Azadirachtin Research Articles

The toxic effects of neem extract and azadirachtin on the brown planthopper, Nilaparvata lugens (Stål) (BPH) (Homoptera: Delphacidae)

Extracts of neem ( Azadirachta indica A. Juss) are used in the developing world for many purposes including management of agricultural insect pests. The effects of different neem extracts (aqueous (NSKEaq), ethanol (NSKEeth) and hexane (NSKEhex)) on mortality, survival and weight of the brown planthopper, Nilaparvata lugens (Stål) (BPH) (Homoptera: Delphacidae) third and fourth nymphal instars were investigated. When fed rice plants treated with neem derivatives in bioassays, the survival of BPH nymphs is affected. Comparisons were made with the pure neem limonoid, azadirachtin (AZA) to ascertain its role as a compound responsible for these effects. AZA was most potent in all experiments and produced almost 100% nymphal mortality at 0.5 ppm and higher concentrations. When higher concentrations were applied, the effects appeared shortly after treatment and mortality was higher. Many insects died after remaining inactive for several days or during prolonged moulting. At lower concentrations, if moulting was achieved, disturbed growth and abnormalities were then likely to occur in the moulting process. Nymphs that were chronically exposed to neem extract showed a reduction in weight (45–60%). The results clearly indicate the simple NSKE (aqueous, ethanolic or both), containing low concentrations of AZA, can be used effectively to inhibit the growth and survival of BPH.

Combined effects of azadirachtin and nucleopolyhedrovirus (SpltNPV) on Spodoptera litura Fabricius (Lepidoptera: Noctuidae) larvae

Abstract The effects of azadirachtin (AZA) and Spodoptera nucleopolyhedrovirus (SpltNPV) on development and mortality of Spodoptera litura Fabricius (tobacco cutworm) were evaluated in the laboratory. The effective concentrations for AZA and SpltNPV were determined and tested as single and combination treatments. AZA and SpltNPV produced synergistic effects on tobacco cutworm mortality in higher dose combination treatment. Combinations of AZA + SpltNPV at 0.25 ppm + 1 × 103 OB and 0.50 ppm + 1 × 106 OB resulted in a significantly higher larval mortality than treatment with either virus/botanical insecticide alone at the corresponding concentrations. When consumed together (AZA and SpltNPV) larvae died significantly faster compared with larvae consuming SpltNPV or AZA. These results suggest that treatments with AZA and SpltNPV at appropriate combinations of concentration levels may result in improved control of tobacco cutworm compared with treatment with either AZA or virus alone.

Soil application of azadirachtin and 3‐tigloyl‐azadirachtol to control western flower thrips, Frankliniella occidentalis (Thysanoptera: Thripidae): translocation and persistence in bean plants

To study the systemic effects of active neem ingredients, the substrate of bean plants was treated with a 170 g kg(-1) azadirachtin (NeemAzal-U; Trifolio-M GmbH, Lahnau, Germany, registration pending). This product was used at a dose rate of 10 mg AZA (azadirachtin a) and 1.2 mg 3-tigloyl-azadirachtol (azadirachtin b) per treated bean plant. Afterwards, the translocation and persistence of AZA and 3-tigloyl-azadirachtol and the effects on western flower thrips, Frankliniella occidentalis (Pergande), were studied. Residues of AZA and 3-tigloyl-azadirachtol from substrates with different contents of organic matter [pure culture substrate (CS), CS-sand mixture] and from various plant parts were quantified by high-performance liquid chromatography-mass spectrometry (HPLC-MS). The dissipation trends of AZA and 3-tigloyl-azadirachtol were similar within the same substrates. A slower decline of both active ingredients was measured with CS than with CS-sand mixture. Residue analysis of the bean plants showed that only small proportions of the initial amounts of AZA and 3-tigloyl-azadirachtol applied to the substrate were present in the plant (0.3-8.1%). Variable amounts of residues of the active components in relation to plant parts and time of analysis indicated a different translocation pattern for the two active ingredients. Higher residues of the active ingredients were found in roots and stems after neem application using CS, whereas higher residues were found in leaves after CS-sand mixture treatments. Mortality of F. occidentalis after NeemAzal-U soil applications reached up to 95% on CS-sand mixture, compared with 86% in CS.

The effects of azadirachtin and nucleopolyhedrovirus on midgut enzymatic profile of Spodoptera litura Fab. (Lepidoptera: Noctuidae)

The effects of azadirachtin (AZA) and nucleopolyhedrovirus (NPV) on midgut enzyme activity in Spodoptera litura Fabricius (Lepidoptera: Noctuidae) (Tobacco cutworm) were evaluated. Gut enzyme activities were decreased by AZA and NPV individually and in combination. When S. litura larvae were fed a diet of castor leaves treated with AZA and NPV in bioassays, gut enzyme—acid phosphatases, alkaline phosphatases, adenosine triphosphatases, and lactate dehydrogenase—activities were decreased. There were statistically significant differences ( P ⩽ 0.05) in enzyme activities between combined and individual treatment. A synergistic effect of botanical insecticides and virus was found when combined in low doses. These effects are most pronounced in early instars. Maximum weight loss (59–72%) occurred, when AZA and NPV were combined.

Efficacy of nucleopolyhedrovirus and azadirachtin on Spodoptera litura Fabricius (Lepidoptera: Noctuidae)

Spodoptera litura Fabricius (Lepidoptera: Noctuidae) is a destructive pest of subtropical and tropical agriculture, and has the potential to be a serious pest of forage crops in Asia. The effects of azadirachtin (AZA) and nucleopolyhedrovirus (NPV) on feeding and mortality of S. litura were evaluated in the laboratory. The nutritional indices were significantly affected. Gross dietary utilization (efficiency of conversion of ingested and digested food) of S. litura was also decreased after treatment with AZA and NPV. Food consumption, efficiency of conversion of ingested and digested food, relative growth rate, and consumption index values declined significantly, but at the same time an increase in approximate digestibility was also observed. Maximum larval weight (fifth instar) loss (68%) occurred, when AZA and NPV were combined. When consumed together (AZA and NPV) larvae died significantly faster compared with larvae that consumed NPV or AZA. Dose and larval instar dependent growth retardation was observed after treatment with NPV and AZA. Upon combination of AZA with NPV, nutritional indices decreased twofold even at reduced concentration of both.

Action of insect growth regulator insecticides and spinosad on life history parameters and absorption in third-instar larvae of the endoparasitoid Hyposoter didymator

Abstract Hyposoter didymator (Thunberg) (Hymenoptera: Ichneumonidae) is an important larval parasitoid of several lepidopteran pests. Under laboratory conditions, the topical toxicity of selected insect growth regulator insecticides, azadirachtin (AZA), diflubenzuron (DFB), methoxyfenozide (MET), pyriproxyfen (PYR), and tebufenozide (TEB), and the naturalyte spinosad (SPIN) was tested against last (third)-instar larvae of this parasitoid up to the maximum field recommended concentration (MFRC). At MFRC, no abnormalities or mortality were scored for MET and TEB (IOBC toxicity classes 1). In contrast, AZA was slightly, PYR moderately, and DFB and SPIN harmful to the parasitoid (IOBC toxicity classes 2, 3, and 4, respectively). These ratings are the result of a reduction in rate of: (i) pupae formation, (ii) pupal mortality, (iii) adult longevity, (iv) parasitism, and (v) adult emergence. In addition, PYR and SPIN caused a direct mortality in the treated larvae. At sublethal concentrations, every insecticide but MET and TEB also affected these life history parameters of the parasitoid. The second part of the study focused on pharmacokinetic parameters important for the toxicological properties of these insecticides. We determined the recovery pattern of 14C-labeled DFB, MET, PYR, and TEB from parasitoid body, larval exuvia, and silken pupal cocoon after topical application to third-instar larvae. For MET, TEF, and PYR, >60% was recovered from the parasitoid body after 24 h, whereas this was only 12% for DFB, which was mostly detected in the silken cocoon (>60%). More than 25% PYR was also detected in the larval exuvia after three days. Data suggest that the silken pupal cocoon and the larval exuvia might play a role in eliminating insecticides. In conclusion, the current results indicated that MET and TEB are harmless to third larvae of H. didymator. In contrast, AZA, DFB, PYR, and SPIN are harmful and their use in IPM should be considered with caution until they are evaluated under field conditions.

Ecological Risk Assessment of Neem-Based Pesticides

A tiered process was used to evaluate the risks of pure azadirachtin (AZA) and two neem-based insecticides (Neemix™ and Bioneem™) on six aquatic animals [crayfish (Procambarus clarkii), white shrimp (Penaeus setiferus), grass shrimp (Palaemonetes pugio), blue crabs (Callinectes sapidus), water fleas (Daphnia pulex), and mosquito larvae (Culex quinquefasciatus)] through short term acute toxicity tests. The risk was calculated using the level of concern endpoints (Q values) and relative hazard index (RHI) for acute and chronic exposure scenarios. The Q values of Neemix™, Bioneem™, and pure AZA derived from acute exposure tests indicated that D. pulex is the only sensitive species to the test pesticides. Furthermore, the RHI values of Neemix™ and Bioneem™ for D. pulex were above the critical limit of 10 indicating that these pesticides may pose a moderate hazard to this species and related crustaceans in acute exposure scenarios. The RHI values of the two pesticides and pure AZA were all below the critical limit of 10 for P. clarkii, P. setiferus, P. pugio, C. sapidus, and C. quinquefasciatus. The aquatic risk assessment process showed that the risk values of tested pesticides did not exceed the criteria, and therefore, no ecological hazard is likely to result from their use.

Analysis of neem oils by LC-MS and degradation kinetics of azadirachtin-A in a controlled environment. Characterization of degradation products by HPLC-MS-MS.

Since it was first isolated, the oil extracted from seeds of neem (Azadirachtin indica A juss) has been extensively studied in terms of its efficacy as an insecticide. Several industrial formulations are produced as emulsifiable solutions containing a stated titer of the active ingredient azadirachtin-A (AZ-A). The work reported here is the characterization of a formulation of this insecticide marketed under the name of Neem-azal T/S and kinetic studies of the major active ingredient of this formulation. We initially performed liquid-liquid extraction to isolate the neem oil from other ingredients in the commercial mixture. This was followed by a purification using flash chromatography and semi-preparative chromatography, leading to (13)C NMR identification of structures such as azadirachtin-A, azadirachtin-B, and azadirachtin-H. The neem extract was also characterized by HPLC-MS using two ionization sources, APCI (atmospheric pressure chemical ionization) and ESI (electrospray ionization) in positive and negative ion modes of detection. This led to the identification of other compounds present in the extract-azadirachtin-D, azadirachtin-I, deacetylnimbin, deacetylsalannin, nimbin, and salannin. The comparative study of data gathered by use of the two ionization sources is discussed and shows that the ESI source enables the largest number of structures to be identified. In a second part, kinetic changes in the main product (AZ-A) were studied under precise conditions of pH (2, 4, 6, and 8), temperature (40 to 70 degrees C), and light (UV, dark room and in daylight). This enabled us to determine the degradation kinetics of the product (AZ-A) over time. The activation energy of the molecule (75+/-9 kJ mol(-1)) was determined by examining thermal stability in the range 40 to 70 degrees C. The degradation products of this compound were identified by use of HPLC-MS and HPLC-MS-MS. The results enabled proposal of a chemical degradation reaction route for AZ-A under different conditions of pH and temperature. The data show that at room temperature and pH between 4 and 5 the product degrades into two preferential forms that are hydrolyzed to a single product over time and as a function of pH change.

Acute Toxicity Assessment of Azadirachtin‐Based Pesticides Using Murine Hybridoma and Oyster Cells

In vitro acute toxicities of azadirachtin‐containing pesticides (Neemix™ and Bioneem™), formulated with neem tree extracts, and pure azadirachtin (AZA), the believed active ingredient, were studied using hybridoma and oyster cells and were compared to results obtained using the standard in vivo Daphnia pulex toxicity assay. Neem‐based pesticides showed relatively high toxicity to both hybridoma and oyster cells at concentrations of 1 μg AZA/mL and higher. The IC50 values for hybridoma cells were 2.15 μg AZA/mL for Neemix™ and 1.67 μg AZA/mL for Bioneem™. Oyster cells had IC50 values of 2.18 μg AZA/mL for Neemix™ and 9.46 μg AZA/mL for Bioneem™. Purified AZA, however, did not appear to be as toxic as the formulations. D. pulex was also more sensitive to neem‐based pesticide exposure than that of pure AZA. The applications and limits of these two in vitro models for testing the acute toxicity of AZA‐based pesticides are discussed in comparison with the in vivo D. pulex test.

Lethal and sublethal effects of a neem-based insecticide on balsam fir sawfly (Hymenoptera: Diprionidae).

Lethal and sublethal effects of Neemix 4.5 EC, a commercial neem preparation, on balsam fir sawfly, Neodiprion abietis (Harris), were determined in the laboratory. Larval mortality of N. abietis increased in a concentration-dependent manner, and lethal time decreased with increasing Neemix 4.5 EC concentration. Fifty percent of the larvae died after 4.6 d at a concentration of 90 ppm azadirachtin (AZA) and 12.3 d at a concentration of 0.08 ppm. Neemix 4.5 EC showed some deterrent effects to feeding site selection on N. abietis larvae at high concentrations, but not at low concentrations. Strong reduction of food intake by N. abietis larvae after exposure to Neemix 4.5 EC was demonstrated by significant reduction of frass production. Larvae fed on Neemix 4.5 EC-treated foliage at 90 ppm AZA produced only 16% as much frass as that produced by larvae fed on control foliage (0 ppm). Neemix 4.5 EC at a concentration of 0.08 ppm AZA retarded larval and pupal development. Sublethal doses significantly reduced pupal weight and adult emergence, although the sex ratio of N. abietis adults was not affected. Results indicate that sublethal effects of Neemix 4.5 EC on N. abietis may contribute greatly to the overall field efficacy.

Effect of neem seed extract on feeding, growth, survival, and reproduction of Diaprepes abbreviatus (Coleoptera: Curculionidae).

A commercially available neem seed extract, Neemix 4.5, containing 4.5% azadirachtin (AZA), was assessed for biological activity against the root weevil Diaprepes abbreviatus (L.), an important exotic insect pest of Florida citrus. Laboratory bioassays against neonatal and 3-wk-old larvae fed sliced carrot treated with Neemix produced dose-dependent larval mortality and reduced fresh weights among survivors of treatments. The weight response was greater than the mortality response for both larval age groups. Neonates treated with 45 mg/liter AZA weighed 60% less than those in the control after 4 wk. Three-week-old larvae treated with 45 mg/ liter AZA weighed 30% less than those in the control after 5 wk. When neonates were exposed to insect diet incorporated with Neemix, reductions in larval survival and weight were observed at concentrations as low as 4.8 mg/liter AZA after 6 wk. Larval growth was inhibited by >97% with 42.9 mg/liter AZA in the diet. A soil drench containing 30 mg/liter AZA reduced the survival and weight gain of neonates added to potted citrus and provided protection to the roots in a greenhouse experiment. A concentration of 90 mg/liter AZA was required to provide protection of citrus roots against 4-wk-old larvae. Reproductive effects were observed when adult weevils were fed foliage treated with Neemix. The numbers of larvae hatching per egg mass were reduced by 27% and 68% at 30 and 90 mg/liter AZA, respectively. These results suggest that Neemix should be further evaluated for use in integrated pest management (IPM) programs of citrus.

Ultrastructural changes in the corpus allatum after azadirachtin and 20-hydroxyecdysone treatment in adult females of Labidura riparia (Dermaptera)

In previous reports, we have shown that the injection of azadirachtin (AZA) as well as 20-hydroxyecdysone (20E) into vitellogenic females of Labidura riparia induces inhibition of vitellogenin synthesis and ovarian development. Juvenile hormone (JH) treatment rescues vitellogenin synthesis and ovarian growth (Sayah et al., 1995, 1996). In this work, we have studied ultrastructural changes of corpus allatum (CA) after injection of 200, 400, and 600 ng of 20E or 1, 3, and 5 μg of AZA. CA cells exhibit signs of inactivity in both AZA and females treated with 20E at doses of 3 μg and 400 ng, respectively. Conspicuous cytological effects consisting of multivesicular bodies with dense contents, abnormally large intercellular spaces comprising myelinic structures, and rare smooth endoplasmic reticula occured in cytoplasm of CA glandular cells in both experimental females. However, the CA ultrastructure of females injected with 20E differs from CA cells of females injected with AZA in having a cytoplasm containing numerous electron-lucent intracellular areas and marked glycogen zones. They also differ in having abundant microtubules and well-developed junctional membranes. At a dose of 600 ng of 20E or 5 μg of AZA, the intensity of the cytotoxic effects is more apparent. CA cells display pycnotic nuclei, spherical mitochondria, large multivesicular bodies, and vacuolization of the cytoplasm. These results are discussed and compared with observations made on other insect species.

Comparative toxicity of two azadirachtin‐based neem pesticides toDaphnia pulex

Azadirachtin (AZA)-based pesticides (Neemix and Bioneem) demonstrated toxicity in 48-h nonrenewal toxicity assays using Daphnia pulex at levels that were comparable with several organophosphate pesticides. The median lethal concentration (LC50) values for the two neem pesticides were found to be 0.028 and 0.033 microl/ml, respectively. The LC50 value for nonformulated (95% pure) AZA was determined to be 0.382 microg AZA/ml. Neemix and Bioneem were exposed to air and northern sky daylight in a light box at 24 and 37 degrees C for 1, 3, 6, and 9 d. Standard 48-h acute toxicity tests were used to determine the effect of aging in these dry environmental conditions. Neemix and Bioneem were also fractionated into volatile and nonvolatile fractions, and the toxicity of each was tested. Compared with Neemix, Bioneem remained toxic longer when exposed to light and air at 37 degrees C, indicating that this pesticide may be less prone to environmental degradation. When fractionated, the nonvolatile fractions for both pesticides exhibited significantly lower LC50 values than the full formulations. These results suggest that, depending on the application rate and environmental fate, AZA-based pesticides may have direct adverse effects on aquatic organisms and that the toxicity and stability of formulated pesticides depend on factors other than only the AZA concentration.

Extraction of azadirachtin A from neem seed kernels by supercritical fluid and its evaluation by HPLC and LC/MS.

A new supercritical extraction methodology was applied to extract azadirachtin A (AZA-A) from neem seed kernels. Supercritical and liquid carbon dioxide (CO(2)) were used as extractive agents in a three-separation-stage supercritical pilot plant. Subcritical conditions were tested too. Comparisons were carried out by calculating the efficiency of the pilot plant with respect to the milligrams per kilogram of seeds (ms/mo) of AZA-A extracted. The most convenient extraction was gained using an ms/mo ratio of 119 rather than 64. For supercritical extraction, a separation of cuticular waxes from oil was set up in the pilot plant. HPLC and electrospray mass spectroscopy were used to monitor the yield of AZA-A extraction.

Oviposition bioassay responses of Culex tarsalis and Culex quinquefasciatus to neem products containing azadirachtin

AbstractNeem (Azadirachta indica A. Juss) products have been shown to exert pesticidal properties against a variety of insect species. In mosquito control programs, such products may have the potential to be used successfully as larvicides. In exploring other advantages of neem products, we studied the oviposition responses of Culex tarsalis Coquillett and Cx. quinquefasciatus Say to two experimental azadirachtin (AZ) formulations, wettable powder Azad™ WP10 (WP) and emulsifiable concentrate Azad™ EC4.5 (EC). Gravid Cx. tarsalis exhibited a distinct preference for the neem suspension of the WP, where significantly more egg rafts were collected from the treatment than from the control. The minimum effective AZ concentration for this activity was 0.5 ppm. The aged suspensions from 1–7 days at 0.5 and 1 ppm AZ were more active in eliciting oviposition responses in Cx. tarsalis than the fresh preparations. This activity of the aged suspensions lasted up to 14 and 21 days at 0.5 and 1 ppm AZ, respectively. Negative ovipositional responses were indicated in the tests of the EC vs. Cx. tarsalis, as well as both neem formulations vs. Cx. quinquefasciatus. In the tests of the EC formulation, significantly less gravid females were trapped by oviposition cups in the treatment than in the control, and in the tests of the WP significantly less egg rafts were collected from the treatment than from the control. The minimum effective concentrations for oviposition avoidance activity were 5 ppm AZ for Cx. tarsalis and 10 ppm AZ for Cx. quinquefasciatus, which lasted up to 1 and 4 days for these two species respectively. Neem products potentially used as mosquito larvicides may have many additional benefits in mosquito control programs, the oviposition modification noted in the current studies is one such example.

Endocrine and neuroendocrine effects of Azadirachtin in adult females of the earwig Labidura riparia

In previous studies we have shown that injection of the insect growth regulator Azadirachtin (AZA) into young vitellogenic females induces inhibition of vitellogenesis in a dose-dependent manner. Juvenile hormone treatment rescues vitellogenin synthesis and ovarian growth. The cytopathological effects on ovaries and fat body are not linked to an inhibition of feeding. In this work we investigated the effects of AZA on the endocrine and neuroendocrine system. Enzyme immunoassay reveals that ovarian ecdysteroid levels are drastically reduced, in a dose-dependent fashion, by AZA. Ultrastructural study indicates that corpus allatum cells exhibit signs of inactivity and degenerative changes after AZA exposure. Using an antibody against allastostatin-3 of Blatella germanica (BLAST-3), we show the appearance of strong immunoreactivity of numerous cells and axons in the brain of AZA-injected females. We conclude that vitellogenesis inhibition by AZA consists of a direct cytotoxic effect as well as a generalized disruption of endocrine and neuroendocrine functions.

Dissipation kinetics of azadirachtin in some forest matrices and its systemic translocation in conifers for spruce budworm control

Field microcosm studies were conducted to investigate the deposition, persistence, translocation and bioactivity of the botanical insecticide azadirachtin‐A (AZ‐A) in young spruce trees. The deposition and persistence of AZ‐A in aspen foliage, forest soil and litter were also studied. AZ‐A residues in the different matrices were monitored at intervals of time after spray application of Neemix® 4.5 formulation at 100 g AZ‐A in 2.3 L/ha. The dissipation half‐lives in the matrices ranged from 16.9 to 33.6 h. AZ‐A was translocated and concentrated in the growing parts (especially the shoots) of spruce trees, after treatment by trunk injection (Tl), basal bark painting (BBP) and foliar painting (FP) of the Neemix 4.5 formulation. Systemically translocated and accumulated AZ‐A residues in the shoots after Tl and FP were found to be effective against spruce budworm (Choristoneura fumiferana Clemens.) larvae. Results suggest that Tl may be a useful way of using neem insecticide to protect high value trees from insect pests.

Formulation selection, and investigation of azadirachtin-A persistence in some terrestrial and aquatic components of a forest environment

Five commercial formulations of azadirachtin-A (AZ-A) Margosan-O®, Azatin-EC®, Neem-EC®, RH-9999 and Neemix® 4.5, were investigated for their volatilization and washoff potential in laboratory studies. Prior to the investigation, RH-9999 (a wettable powder) was mixed with water to provide an end-use formulation containing 35.6 g AZ-A kg -1 , while the remaining four formulations were investigated without dilution. Volatilization and washoff of AZ-A occurred more from white spruce foliage than from wax-coated glass plates. Neem-EC provided the lowest amount of loss, whereas Margosan-O provided the highest. Physical properties and atomization behaviour of the five formulations indicated that Azatin-EC was highly viscous and caused phase separation in droplets collected on glass plates after atomization in a rotary atomizer. RH-9999, despite its low viscosity, caused phase separation in droplets because of the heterogeneity of the wettable powder formulation. Based on the minimum loss of AZ-A due to volatilization and washoff from spruce foliage, and on the minimum potential for phase separation in droplets after atomization in a rotary atomizer, Neem-EC was considered to be the most appropriate choice for use in field studies to investigate environmental persistence and fate of AZ-A in terrestrial and aquatic matrices of a forest ecosystem. The Neem-EC formulation was sprayed at 40 and 80 g AI ha -1 over single spruce trees and on litter and soil plots selected in a mixed-wood boreal forest in Ontario, Canada. In addition, outdoor aquaria containing stream water and sediment were also fortified with the formulation at 400 and 800 g AI ha -1 . Persistence of AZ-A was evaluated using one-year-old spruce needles, current-year shoots, spruce bark, litter, soil, stream water and sediment. The duration of persistence varied from 3 to 6 days in terrestrial matrices, whereas it ranged from 8 to 13 days in water, and 2 to 3 days in sediment. The half-life (DT 50 ) values ranged from 10.7 h (for soil) to 71.6 h (for spruce bark) at the lower dosage rate, and from 18.8 h (for litter) to 76.2 h (for bark) at the higher dosage rate. The DT 50 value for stream water was about 35 h regardless of the dosage rate applied. The data indicated that AZ-A was appreciably labile and short-lived in different forestry matrices, with low DT 50 values.

Effect of additives in the Neem formulations on deposition, volatilization and persistence of Azadirachtin in spruce foliage

Foliar deposits, volatilization and persistence of azadirachtin‐A (AZ‐A) were investigated after application of four spray mixes prepared from a wettable powder (WP) and three emulsifiable concentrate (EC) formulations of neem. They were applied at the dosage rate of 50 g AI in 4 L/ha onto potted spruce seedlings in a laboratory spray chamber. Droplet‐size spectra and deposits were assessed using Kromekote® card/glass plate collection units. Foliar residues [dislodgeable residues (DR), penetrated residues (PR) and total residues (TR)] of AZ‐A and their volatilization were measured by HPLC at different intervals of time up to 60 h after treatment. Differences in the droplet‐size spectra and deposit levels were observed among the four spray mixes due to the influence of additives present in them. Dissipation half‐lives (DT50) of the DR, PR and TR in the foliage were low (range, 19.5 to 38.9 h) and varied according to the residue type and the spray mix used. The DT50 values of the DR were consistently lower (range, 19.5 to 31.9 h) than those of the PR (range, 30.5 to 38.9 h) due to preferential loss of the surface residues. The low DT50 values observed for the DR and TR in the foliage sprayed with the WP spray mix were attributed to the particulate nature of the deposit. AZ‐A volatilized appreciably from the DR rather than from the PR. The variations found in the amounts of AZ‐A volatilized (42 to 58%) and unaccountable (38 to 46%) from the initial TR values in spruce foliage, after 60 h, were attributed to the physical form of the deposits on the target surface and the influences of additives present in the different spray mixes.

Effect of Azadirachtin on vitellogenesis of Labidura riparia (Insect Dermaptera)

This study investigates the effects of the insect growth regulator Azadirachtin (AZA) on the ultrastructure of ovaries and fat body of the earwig Labidura riparia. Ovarian development is severely reduced in AZA-injected females in a dose-dependent manner. Follicles exhibit degenerative changes, separation of follicle cells from the oocyte, and lack of pinocytotic vesicles as of yolk spheres in cortical ooplasm. Adipocytes show fragmented rough endoplasmic reticulum (RER), numerous autophagic vacuoles, multivesicular bodies, osmiophilic lipid droplets, and many large glycogen areas. Gel electrophoresis reveals that vitellogenin is absent from both fat body and hemolymph, and that vitellin is not deposited in the ovary. These pathological effects are not linked to an absence of feeding. The effect of AZA on vitellogenesis is rescuable by Juvenile hormone (JH) treatment. The inhibition of vitellogenesis by AZA is discussed on the basis of its direct cytotoxic effect as well as its interference with the neuroendocrine system.