The residual effect of some products from Neem (Azadirachta indica A. Juss) seeds upon larvae ofBoarmia (Ascotis) selenaria Schiff. in laboratory trials

The effect of Neem on the development ofBoarmia (Ascotis) selenaria Schiff. was investigated. Young leaves of avocado were treated in the laboratory with varying concentrations of Neem seed kernel suspensions or with emulsions of some Neem oil products, and offered to larvae of different size. Furthermore, avocado plants in flower pots were sprayed with Neem suspensions and the residues allowed to age in different locations. Leaves from the different treatments were offered to larvae of various sizes in laboratory tests and the larval weight, pupation rate and mortality were recorded. 0.3% Neem was highly effective on laboratory-treated leaves. A negative correlation was found between the concentration of the Neem suspensions on the one hand and the rate of growth and pupation on the other hand. No activity was found with the emulsion of either Neem extractive or Neem oil obtained from Neem kernels after extraction. In larvae kept for four days on avocado leaves treated with 0.3% Neem suspension, the growth and pupation rate was nearly as low as in larvae fed on treated leaves throughout their life span. The experiment with Neem-sprayed plants kept at different locations indicated the following order of activity of the residues: laboratory ? shade > sun.     

Antifeedant and growth inhibitory effects of calamus oil and neem oil onSpodoptera litura under laboratory conditions

A distillate of the essential oil of the rhizomes of Indian calamus (Acorus calamus L.) and seed oil of neem (Azadirachta indica A. Juss) were evaluated as emulsified foliage sprays againstSpodoptera litura (F.) larvae. A dose-response relationship of both oils was established using food acceptance, feeding ratio, weight gain and larval development as parameters in choice and no-choice tests. Calamus oil at the 0.5% and 1.0% concentrations was effective in both tests in inducing significant feeding deterrence and growth inhibition in early 3rd instar larvae. Neem oil had such an effect only at 2%, particularly in no-choice tests. The significance of these findings in the context of the possible action of calamus oil on gustatory receptors is discussed.     

Effect of Neem extract on Lymantria dispar L. (Lepidoptera: Lymantriidae) and Leptinotarsa decemlineata Say. (Coleoptera: Chrysomelidae)

High antifeedancy and low toxicity of the plant preparation Neem (azadirachtin), on the gypsy moth, Lymantria dispar L. larvae were proved under laboratory conditions. A high index of Neem antifeedancy was confirmed in all tested concentrations.Under laboratory conditions the Neem had satisfying antifeedancy on Leptinotarsa decemlineata Say third instar larvae too. Field experiments also proved that the foliage protective effect of Neem was very significant.Experiments showed that some antifeedants or repellents, such as Neem, should control olygophagous (L. decemlineata, Coleoptera) insects as well as very polyphagous insects such as L. dispar (Lepidoptera).As a biological, plant-derived preparation, Neem could be suitable for integrated pest management (IPM) programs, especially in small orchards, parks, and tree rows in urban environments and on small private gardens.     

Effects of a neem-based insecticide on different immature life stages of the leafminerLiriomyza sativae on tomato

The effects of the commercially available neem insecticide NeemAzal-T/S® (neem) (1% azadirachtin) onLiriomyza sativae (Diptera: Agromyzidae) were studied on tomato in Bangkok, Thailand. Neem emulsions were sprayed on the aerial plant parts at five concentrations (from 0.001% to 0.010% a.i.); different aged residues (0, 1, 3, 5, and 7 days) were tested, in both controlled environment rooms and in greenhouses. Irrespective of the residual age of the application, no significant effects on oviposition or egg hatch were detected. However, neem drastically increased larval mortality at the higher concentrations tested. The L1 and L2 larvae were most susceptible. Irrespective of concentration, larval mortality or inhibition of adult eclosion decreased much faster in greenhouses than in controlled environments with artificial light. Nevertheless, our results suggest that neem has potential to controlL. sativae in netted greenhouses.     

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.     

Toxicity of neem to natural enemies of aphids

Neem [Azadirachta indica (A. Juss.)] seed oil (NSO) applied in three concentrations, 0.5%, 1% and 2%, to potted plants infested with green peach aphid,Myzus persicae (Sulzer), totally prevented adult eclosion of larval coccinellids,Coccinella undecimpunctata L., and reduced adult eclosion of syrphids,Eupeodes fumipennis (Thompson), to 11%, 7% and 0%, respectively, of the controls. Under the same conditions, NSO did not reduce the rate of parasitism ofM. persicae byDiaeretiella rapae (Mcintosh), but emergence of parasitoid adults from aphid mummies collected from treated plants was reduced to 35%, 24% and 0%, respectively, of the controls. Although topical treatment of predator larvae with NSO at concentrations of up to 5% did not affect survival, emergence of parasitoids from mummies dipped in aqueous emulsions of NSO at similar rates was reduced significantly. Under field conditions, sprays of NSO and neem seed extract to plants had no significant impact on the number of aphids parasitized. Although total numbers of predators were reduced, numbers of predators relative to aphid numbers were similar to those in controls. Neem insecticides may be suitable for use in integrated pest management programs, as under field use they appear to be relatively benign to aphid predators and parasitoids.     

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.     

Flufenoxuron, an acylurea insect growth regulator, alters development of Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) by modulating levels of chitin, soluble protein content, and HSP70 and p34 in the larval tissues

The effect of sublethal concentrations, 0.00141% (LC₂₀), 0.00251% (LC₃₀), and 0.0036% (LC₄₀) of a dispersible formulation of an acylurea insect growth regulator, flufenoxuron (Cascade) on certain biochemical parameters in the larvae of Tribolium castaneum was investigated. When neonates were fed on diet treated with sublethal concentrations for 24 h, it was observed that at all concentrations tested, there was a significant reduction in chitin content on the 15th day of development. Total soluble protein content at LC₂₀ and LC₃₀ decreased with increasing age of the larvae. At LC₂₀ and LC₄₀ concentrations there was a progressive increase in the protein: chitin ratio as a function of increase in age of the larvae. SDS-PAGE analysis of the larval tissue extracts indicated gross quantitative changes in some of the protein bands (MW 50-97 kDa). Western blot analysis revealed significant increase in the level of HSP70 in the extracts of larvae fed on LC₃₀ treated diet, on the 7th and 10th day of development in the decreasing order. Conversely, a significant decrease in the hyper-phosphorylated form of p34^cdc2 kinase due to flufenoxuron treatment indicating modulation of cell cycle regulation was observed. Thus, sublethal concentrations of flufenoxuron alter expression of developmentally regulated proteins, HSP70 and p34^cdc2 and chitin formation in a stage-specific manner thereby resulting developmental abnormalities in T. castaneum.     

Polysubstrate monooxygenases (cytochrome P-450) in larvae of susceptible and resistant strains of house flies

The polysubstrate monooxygenases (PSMO or cytochrome P-450) of house fly larvae were studied at the mature larval or “clear gut” stage. Fat body and gut tissues were most efficient in the conversion of aldrin to dieldrin. Microsomal fractions of larval homogenates had the highest PSMO activities, with lower PSMO activities also found associated with mitochondrial fractions. Microsomes from Rutgers (resistant) larvae had higher levels of NADPH:cytochrome c reductase (2×), cytochrome P-450 (2×), aldrin (4×), and heptachlor (9×) epoxidases than microsomes from CSMA (susceptible) larvae. Cytochrome P-450 of Rutgers larvae had an absorption maximum at 449 nm, 2 nm lower than the cytochrome P-450 of CSMA larvae. n-Octylamine spectra showed that the level of high-spin cytochrome P-450 was higher in Rutgers larvae. NADPH:cytochrome c reductase, cytochrome P-450, and aldrin epoxidase were induced by phenobarbital, and Rutgers larvae were shown to be more sensitive to this inducer than CSMA larvae. Induction of larval PSMO by phenobarbital did not affect the expression or the inducibility of PSMO in adults.     

Cry1Ac和Cry2Ab蛋白对大草蛉生长发育及酶活力的影响

为明确转Cry1Ac和Cry2Ab基因棉花对大草蛉的影响,运用Bt蛋白与人工饲料混合的方法,以大于转基因棉花叶片中蛋白含量20倍的剂量饲喂大草蛉初孵幼虫,初步研究了Cry1Ac和Cry2Ab对大草蛉生物学参数和消化酶、解毒酶活性的影响。结果表明：取食含Bt蛋白饲料的大草蛉幼虫的发育历期、体重、蛹重、成虫体重、羽化率等生物学参数与对照相比均没有显著差异;在大草蛉幼虫体内可以检测到含量较高的Cry1Ac和Cry2Ab蛋白,分别为974.92~1 282.39 ng/g鲜重和5 592.62~6 082.92 ng/g鲜重,而在大草蛉成虫体内检测到的Cry1Ac和Cry2Ab蛋白含量非常低,分别为0.29~0.39 ng/g鲜重和50.34~56.71 ng/g鲜重;取食含Bt蛋白的饲料对大草蛉幼虫的类胰蛋白酶、类胰凝乳蛋白酶、氨肽酶和谷胱甘肽-S-转移酶活性有一定的影响,但对大草蛉成虫影响与对照差异不显著。表明大草蛉取食含Bt蛋白的人工饲料后,虽然体内可以检测到一定含量的Bt蛋白,但对大草蛉的生长发育并没有显著的直接不利影响。     

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。     

Inducibility of metabolic insecticide defenses in boll weevils and tobacco budworm caterpillars

Of five cotton allelochemicals, (+)-α-pinene, β-caryophyllene, gossypol, umbelliferone, and scopoletin, all except gossypol induced cytochrome P-450 content, N-demethylation, and epoxidation activities more in tobacco budworm larval midguts than in adult or larval boll weevils. All except gossypol also induced glutathione transferase activity in tobacco budworm larval midguts and adult boll weevils. Microsomal esterase activity was unaffected or suppressed by all five allelochemicals. Soluble esterase activities were unaffected, or induced only in boll weevil larvae. The contrast in insecticide resistance development between the tobacco budworm and the boll weevil may be, in part, related to superior inducibility of cytochrome P-450 and associated activities in the former species in addition to overall higher uninduced activities (26).     

Environmental degradation of the insect growth regulator methoprene: V. Metabolism by houseflies and mosquitoes

The metabolism of methoprene (I, isopropyl (2E,4E)-11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate, trademark Altosid) was investigated in larval mosquitoes and houseflies. The most abundant primary metabolite in third- and fourth-instar Aedes and fourth-instar Culex larvae was the hydroxy ester while the hydroxy acid predominated in third-instar Musca larvae. Biological isomerization of the double bond at C-2 in I (i.e., conversion of (E) to (Z)) was an effective mode of insect detoxication, but these dipterans apparently cannot isomerize the (2Z) isomer of I to methoprene. In general, piperonyl butoxide and triorthocresyl phosphate slightly increased the morphogenetic activity of I in these insects.     

Cell death localization in situ in laboratory reared honey bee (Apis mellifera L.) larvae treated with pesticides

In this study, cell death detected by DNA fragmentation labeling and phosphatidylserine (PS) localization was investigated in the honey bee (Apis mellifera L.) midgut, salivary glands and ovaries after treating larvae with different pesticides offered via an artificial diet. To do this, honey bee larvae reared in an incubator were exposed to one of nine pesticides: chlorpyrifos, imidacloprid, amitraz, fluvalinate, coumaphos, myclobutanil, chlorothalonil, glyphosate and simazine. Following this, larvae were fixed and prepared for immunohistologically detected cellular death using two TUNEL techniques for DNA fragmentation labeling and Annexin V to detect the localization of exposed PS specific in situ binding to apoptotic cells. Untreated larvae experienced ∼10% midgut apoptotic cell death under controlled conditions. All applied pesticides triggered an increase in apoptosis in treated compared to untreated larvae. The level of cell death in the midgut of simazine-treated larvae was highest at 77% mortality and statistically similar to the level of cell death for chlorpyrifos (65%), imidacloprid (61%), myclobutanil (69%), and glyphosate (69%) treated larvae. Larvae exposed to fluvalinate had the lowest midgut columnar apoptotic cell death (30%) of any pesticide-treated larvae. Indications of elevated apoptotic cell death in salivary glands and ovaries after pesticide application were detected. Annexin V localization, indicative of apoptotic cell deletion, had an extensive distribution in the midgut, salivary glands and ovaries of pesticide-treated larvae. The data suggest that the tested pesticides induced apoptosis in tissues of honey bee larvae at the tested concentrations. Cell death localization as a tool for a monitoring the subclinical and sub-lethal effects of external influences on honey bee larval tissues is discussed.     

The residual effect of a neem seed kernel extract sprayed on fodder beet against larvae ofSpodoptera littoralis

The residual effect of a dried methanolic extract of neem (Azadirachta indica A. Juss) seed kernels was investigated with larvae of the Egyptian cotton leafworm,Spodoptera littoralis (Boisd.), in field trials. Fodder beet was sprayed with 1% emulsions of the extract. The treated leaves were collected from the field 1, 3, 5 and 9 days after spraying and offered to the larvae for 48 h. Survival and mean weight of the larvae were recorded 2, 4, 6, 9 and 11 days from the start of exposure to the residues, and rate of pupation was calculated. The extract had a strong antifeedant effect and an intense insect growth-regulating (IGR) effect in field-treated leaves, especially with the 1- and 3-day-old residues. A positive correlation was found between the age of the residues and the mean percent of live larvae, larval weight and pupation rate of the larvae.     

Feeding behaviour as a limiting step in insecticide absorption for the wireworm Agriotes sp. (Coleoptera: Elateridae)

The fipronil control of wireworms field populations (larval Agriotes sp., Coleoptera: Elateridae) is difficult because of the low integumental penetration rates of this insecticide into the target larval body. The main way of insecticide absorption being associated with food, analysis of the larval feeding behaviour is of key importance for designing a strategy of dietary chemical control of those subterranean pests. In this purpose, a standard method for assessing the long-term survival of field specimens in the laboratory was developed together with experimental designs for investigating food searching and choosing by larvae. Larval biological performances under those laboratory conditions were checked monthly. Three criteria were taken into account for measures: the duration of individual survival, the gain in average body weight, and the moulting rhythm. Experimentation revealed that seed flour was highly desired by larvae and that larval feeding choice was affected over short distances only. This suggests the involvement of dietary preference rather than attractivity in the larval feeding behaviour. This implies that, at the operational step, insecticide had better be associated to food, either as a coating on the seeds, or inside preferred baits.     

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 of the chitin synthesis inhibitors,diflubenzuron and its dichloro-analogue,to Spodoptera littoralis larvae

The activities of the chitin synthesis inhibitors, diflubenzuron and PH 60–38, against Spodoptera littoralis larvae were assayed by feeding treated alfalfa or poisoned wheat bran baits, by allowing the larvae to imbibe sucrose-containing aqueous dispersions of the compounds, and by injection into larvae. PH 60–38 was less active than diflubenzuron. On alfalfa, diflubenzuron had to be fed for at least 2 days to prevent formation of normal pupae and emergence of adults. For very big (480–540 mg) larvae, feeding diflubenzuron at concentrations of 50 mg/litre for 2 days or 2.5 mg/litre for 3 days prevented adult emergence. For 200–250 mg larvae, this was achieved by feeding concentrations of 100 mg/litre for 2 days, 5 mg/litre for 3 days or 3.5 mg/litre for 4 days. In all larvae > 150 mg, mortality in feeding experiments occurred in the prepupal or the pupal stage. Only with 30–50 mg and 100–150 mg larvae was there considerable mortality during moults between larval instars, the larvae being unable to liberate themselves from the old larval skins and head capsules. Diflubenzuron incorporated into wheat bran baits at concentrations of from 2.5 to 10 000 μg/g killed approximately 70–90% of the insects. When imbibed, diflubenzuron was much less toxic as a wettable powder than as a liquid formulation but the two formulations were equitoxic when injected into the larvae.     

大灰食蚜蝇幼虫饲养及温室释放试验

在室内用桃蚜、蔷无网蚜、桃粉大尾蚜、菊花蚜虫4种蚜虫饲养大灰食蚜蝇幼虫取得比较满意的结果,幼虫成活率76～80％,蛹羽化率92～94％,子代卵孵化率84～91％。用豌豆修尾蚜饲养,幼虫成活率仅48％,蛹羽化率50％,子代卵孵化率无明显差异。在大灰食蚜蝇幼虫发育期中,前期用桃蚜饲养,后期改用修尾蚜饲养,对幼虫成活率及蛹羽化率仍有影响.食物(蚜虫)充足时,幼虫自残程度低,可以集体饲养。以1:10的益害比释放大灰食蚜蝇3日龄幼虫,防治非洲菊上的蚜虫,3天后蚜虫下降85.9％,效果显著。     

昆虫病原线虫对小地老虎的致病力测定及防治效果

为探讨应用昆虫病原线虫防治小地老虎技术的可行性,采用培养皿滤纸法比较9个品系的昆虫病原线虫对小地老虎幼虫的致病力,同时通过盆栽试验验证了斯氏线虫属小卷蛾斯氏线虫Steinernema carpocapsae NC116品系、芫菁夜蛾斯氏线虫S.feltiae SF-SN品系和异小杆线虫属嗜菌异小杆线虫Heterorhabditis bacteriphora H06品系的保苗效果和防治效果,并测定了苦参碱与NC116品系混用对小地老虎3龄幼虫的联合作用效果。结果表明,NC116品系对小地老虎3龄幼虫致病力最高,其对3龄、4龄和5龄的LD50分别为2.4、7.5、和31.2条/头,表明随小地老虎幼虫龄期递增,其致病力下降。盆栽玉米苗上分别施用线虫100~200条/头时,对玉米保苗效果及对小地老虎3龄幼虫的控制效果依次为NC116H06SF-SN。0.8 mg/L苦参碱和NC116品系混用后,可使小地老虎3龄幼虫死亡率提高109.89%,二者表现增效作用。