Response ofSpodoptera littoralis (Boisd.) andEarias insulana (Boisd.) larvae to azadirachtin and salannin

The effect of azadirachtin and salannin, two triterpenoids isolated from seeds of neem (Azadirachta indica A. Juss), on the feeding response ofSpodoptera littoralis (Boisd.) andEarias insulana (Boisd.) larvae, was investigated. Styropor (foamed polystyrene) lamellae were painted on both sides with a mixture of 5% sucrose with different concentrations of either azadirachtin or salannin dissoled in methanol-water (3∶7). Azadirachtin strongly suppressed feeding inS. littoralis larvae even at 0.001%, whereas salannin showed some antifeedant activity at 0.005% and above. Larvae ofE. insulana were deterred from feeding on azadirachtin-treated lamellae even at 0.005%, whereas salannin was effective only at 0.01% and above. Azadirachtin applied on cotton leaves deterred larvae ofS. littoralis from feeding at all concentrations ranging between 0.001 and 0.02%.     

The effect of diflubenzuron upon larvae of the geometrid Boarmia (Ascotis) selenaria on avocado leaves

Small (70 mg) Boarmia selenaria larvae fed for 4 days on avocado leaves or alfalfa dipped in aqueous diflubenzuron suspensions suffered from severe developmental disturbances. Similar results were obtained with leaves sprayed in an avocado orchard, with which, in addition, the considerable persistence of diflubenzuron under field conditions could be demonstrated. The substance was also active by topical application against large (500–600 mg) B. selenaria larvae, whereas by contact it was only moderately toxic. Some other non-conventional control agents, viz., the antifeedant AC-24055 and several juvenile hormone analogues, were of medium and negligible activity, respectively, against this insect.     

The effect of CME 134 onSpodoptera Littoralis Eggs and larvae

CME 134, a new benzoylphenyl urea chitin synthesis inhibitor, was less active than diflubenzuron and BAY SIR 8514, when tested againstSpodoptera littoralis eggs by a dipping method. AgainstS. littoralis larvae the compound was tested by feeding treated alfalfa, topical application and contact with crystalline residues on glass, followed by observation until the adult stage. With both 200–250 and 360–440-mg larvae 100% mortality was obtained by one-day feeding of alfalfa treated with 0.15 ppm a.i. Topical application to 100- and 200-mg larvae showed CME 134 to be about five and nine times more active than BAY SIR 8514 and diflubenzuron, respectively. These differences were even much greater in the contact tests. Cotton field plots were sprayed with either CME 134 or diflubenzuron formulations, leaves were collected at different intervals and fed for one day toS. littoralis larvae in the laboratory. 0.0009% a.i. CME 134 residues gave complete kill of 30–50-mg larvae after 5 and 20 days, and 86% kill after 28 days of aging. With 0.003 and 0.009% a.i., complete kill was obtained in 200–250-mg larvae until 50 days after spraying.     

The effect of diflubenzuron on Eggs of the vine Moth,Lobesia (Polychrosis) Botrana Den. & Schiff (Lepidoptera: Tortricidae)

The activity of diflubenzuron against eggs ofLobesia botrana Den. & Schiff. was assayed with a dipping procedure, at 22° and 27° C. The compound was much more active at 27°C (LC-50 = 0.0072%) than at 22°C (LC-50 = 0.195%).     

Speed of action and toxicity of acylurea insect growth regulators against Spodoptera exempta (Walk.) and Spodoptera littoralis (Boisd.) larvae: Effect of inter-moult age

Ultra-low volume (ULV) spray bioassays at droplet densities (20–80 drops cm^?2) corresponding to field deposits were used to determine the efficacy of two acylurea insect growth regulators, teflubenzuron (‘Nomolt®’) and flufenoxuron (‘Cascade®’) against three different age groups of third-instar Spodoptera exempta and Spodoptera littoralis larvae. While no response (mortality and/or abnormal moulting with impaired ability to feed) was observed with one-day-old larvae until 48 h after treatment, two-day-old larvae showed some response (27–93% abnormal moulting and 3–10% mortality when larvae were sprayed; 80–100% abnormal moulting and up to 83% mortality when larvae and plants were sprayed) after 24 h and up to 100% mortality at 120 h. Studies with non-feeding, pre-moult larvae showed that cuticular uptake of acylureas was sufficient to affect an appreciable proportion of larvae after 24 h, with up to 100% mortality at 120 h. More detailed studies with pre-moult S. littoralis larvae showed that, while a few larvae which moulted between 1 and 5 h after treatment showed abnormal moulting to the L4 stage, a much larger proportion of larvae which moulted 6–20 h after treatment were affected. Previous studies with S. exempta have shown that different larval instars show similar levels of sensitivity to acylureas. The present work suggests that, while the age distribution within instars has little influence on the end-point mortality of acylureas, it can profoundly modify the speed of response, the majority of larvae within an instar being effectively controlled at the moult following treatment. The significance of these observations in relation to the crop protection activity of acylureas against field populations of Spodoptera larvae is discussed.     

Contact activity of diflubenzuron against Spodoptera littoralis larvae

The effect of diflubenzuron as a residue on glass or applied topically to Spodoptera littoralis larvae was investigated. Diflubenzuron was active as a residue on glass against 100 and 200 mg larvae; the toxicity of residues was identical whether a dispersable formulation or a wettable powder or the technical substance were used. By topical application, diflubenzuron had an ED₅₀ for cumulative percentage mortality up to the adult stage of 004 and 0066 μg/larva for 100 and 200mg larvae respectively. Neither the site of the topical application nor whether the larvae were kept singly or in groups of ten after the treatment had an influence on toxicity. The data indicate that diflubenzuron has contact toxicity to at least one insect species as well as the known stomach poison action.     

The toxicity of diflubenzuron to Oxya japonica (Willemse) and its effect on moulting

A laboratory evaluation of the acute toxicity of diflubenzuron against the final stage of Oxya japonica nymphs showed that it was more effective in preventing the development of the nymphs into the adults when applied topically, than when injected. Histological studies revealed that treated nymphs that subsequently died, before or during ecdysis, suffered from severe endocuticular lesions. Comparable abnormalities and disruption of endocuticular deposition and growth were also seen in nymphs that were treated topically with diflubenzuron at 50 or 200 μg a.i. per insect, although these nymphs appeared normal externally.     

The effect of diflubenzuron feeding on glycosaminoglycan and sulfhemoglobin biosynthesis in mice

Diflubenzuron (Dimilin), an inhibitor of insect chitin synthesis, was fed to mice in concentrations of 50 to 2000 ppm in the diet. The activity of the mammalian hexosamine transferases responsible for glycosaminoglycan (mucopolysaccharide) formation was monitored by measuring the in vivo rate of incorporation of a labeled precursor into hyaluronic acid and chondroitin sulfate of skin. No inhibition of biosynthesis was noted at any concentration; indeed the insecticide appeared to stimulate the production of these compounds. After 3 weeks on the diet, mice eating 1000 and 2000 ppm diflubenzuron showed obvious signs of cyanosis, and sulfhemoglobin was demonstrated in the blood of mice eating more than 200 ppm. The amount of sulfhemoglobin appeared to be related to the dietary insecticide content and the highest level seen was 13% of total hemoglobin in the 2000-ppm group after 31 days. These values returned to normal within 3 weeks when diflubenzuron was removed from the diet.     

The toxic effect of diflubenzuron on Spodoptera littoralis eggs and on their respiration

The mortality rates of Spodoptera littoralis eggs of different ages when dipped in aqueous dilutions of a liquid diflubenzuron formulation were investigated. Eggs 2-3 days old were less sensitive than younger eggs, 0-1 day old. The respiratory rate of eggs 0-1 day old, 1 or 2 days after treatment, was the same as in the control. Eggs in the control hatched on the next day; on this day, respiration in the treated eggs decreased rapidly, but further decrease was then gradual and the base line was not reached until 11 days after treatment, or 9 days after the maximum respiratory rate had been obtained. The curve with diflubenzuron resembled that obtained with parathion.     

Response ofTearias Insulana boisd. larvae to neem (Azadirachta Indica a. juss) Kernel extract

The effect of Neem on the development ofEarias insulana Boisd. was investigated. Different concentrations of an aqueous extract of Neem kernels were incorporated in a semi-synthetic diet and larval growth, pupation rate and mortality of larvae were recorded. Neem was highly effective against 7-8- and 12-day-old larvae at all concentrations ranging between 0.075 and 1%. The systemic activity of Neem was investigated by standing cotyledons of a glandless cotton strain with their petioles in different concentrations of Neem. After 72 hours feeding at all concentrations tested, larval weight was about half that on the untreated leaves.     

Laboratory selected resistance to diflubenzuron in larvae of aedes aegypti

The variation in tolerance to diflubenzuron [1-(4-chlorophenyl)-3-(2,6-difluorobenzoyl)urea] was examined in fourth instar larvae of seven strains of Aedes aegypti, some of which were resistant to DDT and permethrin. The difference between the least and the most tolerant to diflubenzuron was approximately two-fold. There was no correlation with resistance to the other insecticides. A DDT-resistant strain (T8) was selected 10 times (during 12 generations) with diflubenzuron. The LC₅₀ to diflubenzuron had increased 3.3 times by the S₈ generation but there was no further increase in later generations despite further selection. Associated with this increase, a marked decrease in resistance to DDT was observed but no change in permethrin tolerance. A genetically enriched strain (Hotchpotch) was synthesised from 35 strains of different geographic origin and crossed to the selected T8 strain before subsequent generations were selected five times with diflubenzuron. This procedure resulted in an 8 to 12-fold increase in the LC₅₀ value over that for unselected T8, accompanied by a decrease in the slope of the log dose against probit mortality line.     

The inhibition of chitin synthesis in spodoptera littoralis larvae by flufenoxuron,teflubenzuron and diflubenzuron

The chitin precursor [¹⁴C] N-acetylglucosamine injected into the haemolymph of Spodoptera littoralis (Boisduval) larva was incorporated into the chitin exponentially with time. When caterpillars were injected with precursor at the commencement of feeding on acylurea-treated leaf discs, flufenoxuron, teflubenzuron and diflubenzuron were found to be equally effective inhibitors of chitin synthesis, measured after 21 h. The dose response curves by feeding are not parallel, indicating that the relative potency of the compounds will vary across the dose range. When chitin precursor was injected simultaneously with topically applied diflubenzuron, flufenoxuron or teflubenzuron, all three acylureas were found to be equally effective as inhibitors of chitin synthesis when measured after five hours. The I₅₀values (50% inhibition of chitin synthesis) were not significantly different; average 600 ng, compared with LD₅₀values (50% lethal dose) of ～ 13 ng for flufenoxuron and teflubenzuron but 130 ng for diflubenzuron (topical application). Injection of precursor 24 h after topical application of insecticide gave an I₅₀value which had dropped 670- and 150-fold for flufenoxuron and teflubenzuron respectively but only 20-fold for diflubenzuron. It is postulated that the reason for the low increase in diflubenzuron effectiveness with time was due either to less diflubenzuron than flufenoxuron reaching the site of action, or more probably, a faster rate of metabolism and excretion for diflubenzuron. The lower toxicity of diflubenzuron compared with flufenoxuron and teflubenzuron may not be due to any inherent differences in biochemical effectiveness, but rather to different penetration/metabolism properties.     

Development of resistance and cross-resistance in fenvalerate and deltamethrin selected strains of Spodoptera littoralis (Boisd.)

Resistances of 33- and 27-fold to the pyrethroids fenvalerate and deltamethrin, respectively, have been induced in larvae of the Egyptian cotton leafworm Spodoptera littoralis (Boisd.) by selection of two subcolonies of a field population in the laboratory. Selected strains showed relatively high levels of resistance (11- to 36-fold) to all the tested pyrethroids, moderate levels of cross-resistance (3- to 13-fold) to DDT and the carbamate aminocarb, and a slight increase in tolerance levels to endrin and lindane. The pyrethroid-resistant strains, by contrast, manifested no cross-resistance to the organophosphorus compounds that were tested. Moreover, a negative crossresistance was observed in the pyrethroid-resistant strains to dicrotophos, methamidophos and the carbamate methomyl.     

Effects of ingested diflubenzuron on the longevity and the peritrophic membrane of adult mealworms (Tenebrio molitor L.)

When fed to adults of Tenebrio molitor, diflubenzuron reduced the longevity, the weight of the adults and the thickness of the post-ecdysial adult cuticle. It also affected the production of the peritrophic membrane. The loss of weight and the decrease of longevity of the treated adults may have been because of alterations to the peritrophic membrane, caused by the inhibition of chitin biosynthesis by diflubenzuron.     

Effect of Margosan-O on the development ofEarias insulana

The effect of Margosan-O, a formulation of a natural insecticide obtained from the neem tree (Azadirachta indica A. Juss), on the development of and the penetration rate of larvae of the spiny bollworm,Earias insulana (Boisd.), into cotton bolls was investigated by (i) incorporating Margosan-O into a semisynthetic diet and recording mortality until the adult stage; and (ii) spraying leaves and bolls with, or standing cotton seedlings or twigs of cotton with one boll per twig in aqueous dilutions of Margosan-O in an attempt to protect leaves or cotton bolls from feeding or penetration, respectively, by the larvae. Margosan-O was found to be effective when incorporated into the semisynthetic diet, even at low concentrations, especially when it was offered throughout the larval life span. Survival rate and average weight of the larvae were much lower on leaves treated with 0.5% or 0.2% Margosan-O, or exposed to its systemic effect at 0.5% or 0.25%, than on control leaves. Larval penetration into Margosan-O-sprayed bolls at concentrations of 0.01– 0.5% wasca 30–45% of that into control bolls. Margosan-O was active also as systemic treatment: onlyca 35% of the larvae penetrated the 0.25%-treated bolls compared with the untreated ones.     

The effect of BAY SIR 8514, diflubenzuron,and Hercules 24108 on growth and development ofTribolium confusum

Phytoparasitica - The effect of BAY SIR 8514, diflubenzuron and Hercules 24108 on larval growth, pupation and emergence ofTribolium confusum was determined. BAY SIR 8514 was about twice as active...     

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.     

Beetle-specific Bacillus thuringiensis Cry3Aa toxin reduces larval growth and curbs reproduction in Spodoptera littoralis (Boisd.)

Application of Bacillus thuringiensis tenebrionis (Bt) and expression of the Bt protein Cry3Aa in genetically modified crops are used for targeted control of the Colorado potato beetle Leptinotarsa decemlineata (Say). The Cry3A proteins are selectively toxic for the beetles but the present study describes effects of Cry3Aa on the Egyptian armyworm, Spodoptera littoralis (Boisduval). Cry3Aa expressed in potatoes or added to an agar-base wheat-germ diet reduced the growth of S. littoralis caterpillars and the fertility of adults. The effect of 1.4 mg kg(-1) Cry3Aa in potato leaves was comparable with that of 3.3 mg kg(-1) in the diet. This difference in activity was correlated with better digestibility and higher conversion efficiency of the diet that also supported higher reproduction rate: S. littoralis grown on the potatoes reached a similar size to those on the diet but laid only 702 instead of 1077 eggs per female. Cry3Aa consumption reduced body growth as a consequence of lower food intake without significantly affecting food digestibility and the conversion efficiency of nutrients. The 11% and 5% body weight reductions caused by 1.4 mg kg(-1) Cry3Aa in potatoes and 3.3 mg kg(-1) in the diet, respectively, were associated with 74% and 65% reduction in the number of progeny; S littoralis grown on a diet with 9.1 mg kg(-1) Cry3Aa were 10% smaller and produced no viable progeny. These data suggest that the curtailment of reproduction was not caused by a general shortage of nutrient reserves but by a more direct Cry3Aa effect on the reproduction process.     

Mating,longevity, fertility and fecundity of the cotton leaf-worm,Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae)

The mechanisms adopted to ensure mating success, fertility and fecundity ofS. littoralis were studied. The male moths usually mate only once a night but they mate repeatedly during most of their life span, and practically all their spermatophores are equally efficient for inseminating a female. Although females are also multiple maters, one spermatophore is sufficient for a female to lay fertile eggs throughout her life with no reduction in potential fecundity. While a male may mate 5.3 times, on the average, a female may mate only 2.3 times. As a pronounced decrease in pheromone release occurs after mating, a repeat mating by a female will occur only after most neighboring competitive virgin females have been inseminated. A male may mate therefore with 5.3 different females, all of which will be thoroughly inseminated. The female moth is ready to mate just after emergence and at any time throughout her life. If mating is delayed the preoviposition period is greatly prolonged and eggs are saved to be laid later, after mating, as fertile eggs. Delay in egg laying due to delayed insemination does not result in reduced fecundity as it is compensated for by prolonged longevity. Even if mating is greatly delayed and occurs after egg laying has already begun, the potential reduction in reproduction is not very great, due to the low daily egg-laying rate and the increased longevity of unmated females. It appears unreasonable to expect any tendency toward reduction of egg production with a decrease in the ratio of adult males in a population; for this purpose the control of males must be essentially complete. Therefore, a behavioral control program by mass trapping ofS. littoralis males is unlikely to be economically feasible.