Azadirachta indica (neem) leaf dietary effects on the immunity response and disease resistance of Asian seabass,Lates calcarifer challenged with Vibrio harveyi

The present study was aimed to address the possible evaluation of Azadirachta indica (neem) leaf-supplemented diets on innate immune response in Asian seabass, Lates calcarifer fingerlings against Vibrio harveyi infection. Fish were fed for two weeks diets containing six graded levels of neem leaf at 0 g, 1 g, 2 g, 3 g, 4 g and 5 g per kg feed. Fish fed neem leaf-supplemented diet displayed significant differences (p < 0.05) in weight gain, specific growth rate (SGR) and feed conversion ratio (FCR) compared to the control group fed without neem leaf-supplemented diet. Various innate immune parameters were examined pre-challenge and post-challenge. Fish was injected intraperitoneally with a lethal dose of V. harveyi containing 10⁸ cells mL^?1. Supplementation of neem leaf diet significantly increased phagocytic activity, superoxide anion production, serum lysozyme, serum bactericidal activity, serum anti-protease activity throughout the experimental period when compared with the control group. Dietary doses of neem leaf diet significantly influenced the immune parameters, haematological parameters and blood biochemical indices of treated fish. The results suggested that fish fed neem leaf-supplemented diet improved the immune system and increased survival rate in L. calcarifer fingerlings against V. harveyi infection.     

Nematode inhibiting organic materials and a strain of Trichoderma harzianum effectively manages Meloidogyne incognita in Withania somnifera fields

The usefulness of Trichoderma harzianum was tested along with farmyard manure, cow urine, neem oil seed cake, and vermicompost separately and in combination to manage Meloidogyne incognita in Withania somnifera. A treatment combination of nematode inhibitory vermicompost and T. harzianum was found to be most effective against M. incognita.     

Effects of neem leaf extract on production of aflatoxins and activities of fatty acid synthetase,isocitrate dehydrogenase and glutathione S-transferase in Aspergillus parasiticus

The relationship between the activities of 3 cytosolic enzymes with aflatoxin biosynthesis in Aspergillus parasiticus cultured under different conditions has been investigated in order to find out the role of each enzyme in aflatoxin biosynthesis. Basically the activity of isocitrate dehydrogenase (IDH) was higher in non-toxigenic strains as compared to its counterpart toxigenic fungi (p < 0.05). In contrast, the activities of fatty acid synthase (FAS) as well as glutathione S-transferase (GST) were higher (P < 0.05) in toxigenic strains than that of the non-toxigenic fungi. Aflatoxin production was inhibited in fungi grown in presence of various concentrations of neem leaf extract. Aflatoxin was at its lowest level (>90% inhibition) when the concentration of neem extract was adjusted to 50% (v/v). No significant changes in FAS and IDH activities were observed when aflatoxin synthesis was under restraints by neem (Azadirachta indica) leaf extract. During a certain period of time of culture growth, when aflatoxin production reached to its maximum level, the activity of FAS was slightly induced in the toxigenic strains fed with a low concentration (1.56% v/v) of the neem leaf extract. At the time (96 h) when aflatoxin concentration reached to its maximum levels, the activity of GST in the toxigenic fungi was significantly higher (i.e., 7–11 folds) than that of non-toxigenic strains. The difference was highest in mycelial samples collected after 120 h. However unlike FAS and IDH, GST was readily inhibited (67%) in mycelia fed with 1.56% v/v of the neem extract. The inhibition reached to maximum of 80% in samples exposed to 6.25–12.5% of the extract. These results further substantiate previous finding that there is a positive correlation between GST activity and aflatoxin production in fungi.This revised version was published online in October 2005 with corrections to the Cover Date.     

Effect of vermicompost on nutrition and intermediary metabolism of Colorado potato beetle,Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae)

Colorado potato beetle (CPB) is the important pest of potato throughout the world. The study showed the effects of vermicompost on nutritional indices, digestive enzyme activities and intermediary metabolism of the larvae and the adults of CPB. Vermicompost affected significantly the efficiency of conversion of ingested food (ECI) and efficiency of conversion of digested food (ECD) in addition to activities of carbohydrases and proteases in both larvae and adults. Amount of total phenol compounds increased in the leaves of the potatoes grown on the soil containing 30% of vermicompost compared to control and it were amended by 15% of vermicompost. In case of intermediary metabolism, activities of aspartate aminotransferase and γ-glutamyl transferase showed no significant differences in the control and the treated larvae, but those were fluctuated upper and lower when 15% of vermicompost was added into growth plots. The amount of low-density lipoproteins (LDL) and high-density lipoproteins (HDL) in the larvae of CPB showed no significant differences among treatments. However, the amounts of LDL, HDL, glycogen and protein of the adults significantly increased using 30% of vermicompost. Results of the current study clearly revealed significant effects on some physiological processes in CPB fed on the plants grown in the different vermicompost treatments.     

Effects of neem-based insecticides on beet armyworm （Lepidoptera： Noctuidae）

Abstract Three commercial neem [ Azadirachta indica A. Juss (Meliaceae)]-based insecticides, Agroneem, Ecozin, and Neemix, and a non-commercial neem leaf powder, were evaluated for oviposition deterrence, antifeedant effect on larvae, and toxicity to eggs and larvae of the beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), on cotton leaves in the laboratory. Oviposition deterrence in no-choice, and two- and fivechoice assays, was observed for the neem-based insecticide treatments when compared with a non-treated control. Neem-based insecticides also deterred feeding by beet armyworm larvae. Direct contact with neem-based insecticides decreased the survival of beet armyworm eggs. Survival of beet armyworm larvae fed for 7 days on leaves treated with neembased insecticides was reduced to 27, 33, 60, and 61% for neem leaf powder, Ecozin, Agroneem, and Neemix, respectively. Possibilities for adoption of neem-based insecticides in commercial cotton for beet armyworm control are discussed.     

<Emphasis Type="Italic">Bacillus subtilis</Emphasis> and Vermicompost Suppress Damping-off Disease in Psyllium through Nitric Oxide-Dependent Signaling System

Fusarium oxysporum is one of pathogens causing the damping-off disease of Plantago psyllium in Iran. A greenhouse experiment was conducted to assess the effect of Bacillus subtilis and vermicompost singly and in combination on control of Fusarium–induced damping-off in psyllium. The results showed that vermicompost or B. subtilis, significantly increased the growth of psyllium seedlings and both were effective biocontrol agents against F. oxysporum. Among treatments at least damping-off incidence was recorded in combination of 50% vermicompost and B. subtilis. Results for the first time exhibited that vermicompost as well as B. subtilis induced systemic resistance through nitric oxide (NO) signaling and their combined application further than their individual treatments induced development of plant defense related enzymes including β-1,3-glucanase (GLU), phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO) and the activities of antioxidant enzymes (ascorbate peroxidase, catalase, superoxide dismutase and peroxidase) and also more effectively reduced lipid peroxidation in psyllium leaves. These findings suggested potential of B. subtilis in promoting plant growth as well as inducing systemic resistance in the host plants, was enhanced by vermicompost application.     

Effect of methanolic extracts of neem (Azadirachta indica A. Juss) and bakain (Melia azedarach L) seeds on oviposition and egg hatching of Earias vittella (Fab.) (Lep., Noctuidae)

Abstract: The effects of methanolic extracts of neem ( Azadirachta indica ) and bakain ( Melia azedarach ) seeds on the oviposition behaviour and hatchability of eggs of Earias vittella were investigated under laboratory conditions. Treatments included 2, 4, 6, 8 and 10% concentrations of both extracts to compare with nimbecidine and untreated control. When able to choose, the adults preferred to lay a greater number of eggs on the untreated portion of the oviposition substrate (muslin) compared with the extract-treated portion. However, under no-choice conditions, higher concentrations of extracts caused proportionate reductions in the number of eggs laid. The extracts also manifested repellent activity by reducing the number of eggs laid even when the ovipositing females were not in direct contact with the extracts. Eggs laid on extract-treated oviposition substrate exhibited reduced hatching and marked adverse effects on hatching were noticed when the eggs were dipped in different concentrations of extracts. Adults fed on an extract-containing sucrose diet laid significantly fewer eggs with poor hatching. There was no egg laying when the moths were fed on a sucrose diet containing 6, 8 and 10% neem and 10% bakain extracts.     

Effects of neem seed derivatives on behavioral and physiological responses of the Cosmopolites sordidus (Coleoptera: Curculionidae)

Both in a choice and multi-choice laboratory tests, fewer adults of the banana root borer, Cosmopolites sordidus (Germar), settled under the corms of the susceptible banana "Nakyetengu" treated with 5% aqueous extract of neem seed powder or cake or 2.5 and 5% emulsified neem oil than on water-treated corms. Feeding damage by larvae on banana pseudostem discs treated with 5% extract of powdered neem seed, kernel, or cake, or 5% emulsified neem oil was significantly less than on untreated discs. The larvae took much longer to locate feeding sites, initiate feeding and bore into pseudostem discs treated with extract of powdered neem seed or kernel. Few larvae survived when confined for 14 d on neem-treated banana pseudostems; the survivors weighed two to four times less than the larvae developing on untreated pseudostems. Females deposited up to 75% fewer eggs on neem-treated corms. In addition, egg hatching was reduced on neem-treated corms. The higher the concentration of neem materials the more severe the effect.     

Bioactivity of neem, Azadirachta indica, against spittlebug Mahanarva fimbriolata (Hemiptera: Cercopidae) on sugarcane

The effect of neem, Azadirachta indica A. Juss, on some biological parameters of Mahanarva fimbriolata (Stil) (Hemiptera: Cercopidae) was studied in the laboratory by using NeemAzal-T/S, Nimkol-LS, and an aqueous neem seed extract. Initially, the LC,, was estimated for nymphs. Later, nymphs fed on sugarcane, Saccharum officinarum L., roots were sprayed with the respective LC,, for each product. The adults were maintained in cages on sugarcane plants sprayed at the base with the maximum rate recommended commercially for the crop (3 liter/ha). Moistened cotton discs surrounding the base of the plant were used as oviposition substrates. The LCso values estimated for NeemAzal, Nimkol, and aqueous extract were 0.014, 0.225, and 0.611%, respectively. There was a reduction in spittlebug longevity, regardless of sex, in relation to the control. Males exposed to the neem products, and aqueous extract showed longevity reductions of approximately 50%, whereas for females the reductions were 55-60%. The neem products and extract reduced fecundity by 75-85%. Morphological and physiological changes were observed in 9% of the eggs from individuals submitted to NeemAzal. Neem-based products, especially NeemAzal, have potential for the control of M. fimbriolata.     

Pig manure vermicompost as a component of a horticultural bedding plant medium: effects on physicochemical properties and plant growth

This experiment was designed to characterize the physical, chemical and microbial properties of a standard commercial horticultural, greenhouse container, bedding plant medium (Metro-Mix 360), that had been substituted with a range of increasing concentrations (0%, 5%, 10%, 25%, 50% and 100% by volume) of pig manure vermicompost and to relate these properties to plant growth responses. The growth trials used tomatoes (Lycopersicon esculentum Mill.), grown in the substituted media for 31 days under glasshouse conditions, with seedling growth recorded in 20 pots for each treatment. Half of the tomato seedlings (10 pots per treatment) were watered daily with liquid inorganic fertilizer while the other half received water only. The percentage total porosity, percentage air space, pH and ammonium concentrations of the container medium all decreased significantly, after substitution of Metro-Mix 360 with equivalent amounts of pig manure vermicompost; whereas bulk density, container capacity, electrical conductivity, overall microbial activity and nitrate concentrations, all increased with increasing substitutions of vermicompost. The growth of tomato seedlings in the potting mixtures containing 100% pig manure vermicompost was reduced, possibly as a result of high soluble salt concentrations in the vermicompost and poorer porosity and aeration. The growth of tomato seedlings was greatest after substitution of Metro-Mix 360 with between 25% and 50% pig manure vermicompost, with more growth occurring in combinations of pig manure vermicompost treated regularly with a liquid fertilizer solution than in those with no fertilizer applied. Some of the growth enhancement in these mixtures seemed to be related to the combined effects of improved porosity, aeration and water retention in the medium and the high nitrate content of the substrate, which produced an increased uptake of nitrogen by the plant tissues, resulting in increased plant growth. When the tomato seedlings were watered daily with liquid inorganic fertilizer, substitution of Metro-Mix 360 with a very small amount (5%) of pig manure vermicompost resulted in a significant increase in the growth of tomato seedlings. Such effects could not be attributed solely to the nutritional or physical properties of the pig manure vermicompost. Therefore, it seems likely that the pig manure vermicompost provided other biological inputs, such as plant growth regulators into the container medium, that still need to be identified fully.     

Azadirachta indica metabolites interfere with the host-endosymbiont relationship and inhibit the transmission of potato leafroll virus by Myzus persicae

The effects of neem (Azadirachta indica A. Juss) seed kernel extracts (NSKE) and azadirachtin on the ability of Myzus persicae (Sulz.) to transmit potato leafroll luteovirus (PLRV) was studied. Moreover, it was investigated whether treatments with these compounds would exert an effect on larval growth and mortality, and on the aphid intracellular symbionts. Endosymbiotic bacteria play an essential role in the performance of aphids, and in luteovirus transmission by aphids. NSKE and azadirachtin were offered to one-day-old M. persicae nymphs via a membrane feeding system. The neem metabolites displayed a 100% mortality at doses higher than 2560 ppm. At intermediate doses, ranging between 320 and 2560 ppm, larval growth and mortality were affected in a dose-dependent manner. The transmission of PLRV by M. persicae was inhibited by 55–90%. The endosymbiont population of the aphid was clearly affected by a treatment with neem metabolites as the release of their most abundant protein, Buchnera GroEL, into the haemocoel of the aphid was inhibited. Moreover, morphological aberrations on the bacterial endosymbionts were observed in aphids which fed on 2560 ppm of azadirachtin. At doses lower than 160 ppm of NSKE or azadirachtin, the endosymbiont population of M. persicae, and mortality, growth and feeding behaviour were similar to that of the untreated groups of aphids. However, PLRV transmission was still inhibited by 40–70%. The possible targets of the neem metabolites in the aphid are discussed.     

Aberrations caused by neem oil odour in green leafhopper feeding on rice plants

Using an electronic device and a lignin-specific dye, the feeding behaviour of the green leafhopper,Nephotettix virescens (Distant), (Homoptera: Cicadellidae) was monitored on rice plants kept in an arena permeated with the odour of neem (Azadirachta indica A. Juss) seed oil. The garlicky odour of neem oil disrupted the normal feeding behaviour of the leafhopper. Phloem feeding byN. virescens on rice plants kept in an arena permeated with odour of 6, 12, or 25% neem oil was significantly reduced compared with that on control plants kept in an arena with paraffin oil or 1.66% aqueous Teepol solution. Reduced intake from phloem was associated with a significant increase in the insect's probing frequency and an increase in durations of salivation and xylem ingestion.

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Alterations du comportement sur riz de Nephotettix virescens, dues à l'odeur d'essence de neem (Azadirachta indica)

Résumé De nombreux essais en plein champ à l'I.R.R.I. ont révélé une réduction du T.V. (tungro virus) transmis par (Nephotettix virescens) sur des parcelles de riz sensible non-traitées, entourées de parcelles pulvérisées périodiquement avec de l'essence de graines de neem (Azadirachta indica).L'essence de neem a une forte odeur d'ail, perceptible à distance. Nous avons examiné l'effet de l'odeur d'essence de neem sur le comportement alimentaire deN. verescens, grâce à un dispositif électronique et un colorant spécifique de la lignine. Sur du riz maintenu dans une enceinte imprégnée d'odeur d'essence de neem, nous avons constaté une diminution de la consommation de phloème, une augmentation significative de la fréquence des piqûres et une augmentation de temps de salivation et d'absorption de xylème. Ces altérations du comportement alimentaire provoquées par l'odeur d'essence de neem peuvent expliquer la diminution de la fréquence du T.V. spécifique du phloème dans des champs de riz non-traités, entourés de champs pulvérisés avec de l'essence de neem.

     

Isolation and characterization of microsatellite markers in Indian neem (Azadirachta indica var. indica A. Juss) and cross-amplification in Thai neem (A. indica var. siamensis Valenton)

We developed eight polymorphic microsatellite loci in Indian neem (Azadirachta indica var. indica) and cross-amplified in closely related species Thai neem (A. indica var. siamensis). The number of alleles per locus in Indian neem and Thai neem ranged from 3 to 9 and 3 to 9, respectively. Average observed and expected heterozygosities in Indian neem and Thai neem were 0.63 and 0.70 and 0.61 and 0.65, respectively. Two loci exhibited significantly fewer heterozygotes than expected under Hardy–Weinberg equilibrium. These microsatellites may provide a useful tool for population genetics to establish conservation and management strategy.     

Effect of vermicompost and cucumber cultivar on population growth attributes of the melon aphid (Hemiptera: Aphididae)

Worldwide, the developing industry of cucumbers (Cucumis sativus L.) grown in greenhouses is threatened by damage from sucking pests, especially aphids. Among these, the melon aphid, Aphis gossypii Glover (Hemiptera: Aphididae), is the most serious. We tested the effect of two cucumber cultivars ('Royal' and'Storm') and three vermicompost concentrations (0 [control], 20, and 30%) in field soil on the development and fecundity rates of A. gossypii, by using a randomized complete block design with four replicates as a factorial experiment. The developmental times of nymphs reared on plants grown into the three vermicompost concentrations ranged from 5.5 (0%) to 8.7 (30%) d (on Storm) and from 4.3 (0%) to 7 (30%) d (on Royal). The developmental time of melon aphid's nymphs was greatest on plants grown in the culture medium with 30% vermicompost rate and least on plants reared in the soil without vermicompost. The intrinsic rate of natural increase (r,,) of A. gossypii ranged from 0.204 d(-1) on plants grown in the soil amended with 30% vermicompost rate (on Storm seedlings) to 0.458 d(-1) on plants grown in the soil without vermicompost (on Royal seedlings). Accordingly, our findings confirm that a combination of a low level of vermicompost and a partially resistant cucumber cultivar might play an important role in managing this aphid on cucumbers in greenhouses.     

Impact of Chemical,Organic and Bio-Fertilizers Application on Bell Pepper, <Emphasis Type="Italic">Capsicum annuum</Emphasis> L. and Biological Parameters of <Emphasis Type="Italic">Myzus persicae</Emphasis> (Sulzer) (Hem.: Aphididae)

Myzus persicae (Sulzer) is a polyphagous aphid that causes chlorosis, necrosis, stunting, and reduce growth rate of the host plants. In this research, the effects of Zinc sulfate and vermicompost (30%), Bacillus subtilis, Pseudomonas fluorescens, Glomus intraradices, G. intraradices × B. subtilis, and G. intraradices × P. fluorescens compared to control was investigated on the growth characters of Capsicum annuum L. and biological parameters of M. persicae. Different fertilizers caused a significant effect on growth characters of C. annuum and biological parameters of M. persicae. The highest plant growth was observed on Zinc sulfate and B. subtilis treated plants, and the lowest was on control. Increase in the amount of specific leaf area (SLA) (0.502 mm² mg^?1) was significantly higher in the B. subtilis than other fertilizer treatments. The longest (10.3 days) and the shortest (5.3 days) developmental times of M. persicae nymphs were observed on 30% vermicompost and Zinc sulfate treatments, respectively. The lowest adult longevity periods of M. persicae (11.2 and 11.3 days) were observed on G. intraradices × B. subtilis and 30% vermicompost treatments, respectively, and the longest ones (16.4 days) on Zinc sulfate. The highest rate of nymphal mortality and the lowest amount of nymphal growth index (NGI) were recorded on 30% vermicompost. The nymphs reared on Zinc sulfate treatment had the lowest rate of nymphal mortality and the highest amount of NGI. Thus, amending the soil with 30% vermicompost had a significantly negative effect on the biological parameters of M. persicae that can be used as an ecological control tactic for this pest.     

SYSTEMIC INSECTICIDAL ACTION OF AZADIRACHTIN, NEEM SEED AND CHINABERRY SEED EXTRACTS APPLIED AS SOIL DRENCHES TO POTTED PLANTS

Abstract  The systemic insecticidal activities of: 1) azadirachtin, a potent insect antifeedant and growth regulator isolated from seeds of the neem tree, Azadirachta indica , 2) a neem seed extract containing 0. 30 quantitative fraction of azadirachtin, and 3) a methanolic extract of seeds from the chinaberry tree, Melia azedarach , a close relative of neem were determined. Soil drenches of azadirachtin or the neem seed extract at 1 and 2. 5 mg to potted potato ( Solanum tuberosum ) and tomato ( Lycopersicon esculentum ), remarkably retarded the growth of Manduca sexta larvae and strongly disrupted the pupation of Leptinotarsa decemlineata larvae, respectively. However, five to ten times as much azadirachtin or neem seed extract was needed to reduce the amount of tomato and potato foliage consumed. On sorghum ( Surghum biocolor ), much higher quantity (≥25 mg) of azadirachtin or the neem seed extract was necessary to slow the population growth of Schizaphis graminum , and prolong the survival of treated plants. But methanolic extracts of chinaberry seeds ( M. azedarach ) from two sources (Guangdong, PRC and Hurricane, Utah, USA) at 150 mg per pot were ineffective in reducing aphid population growth and preventing plant damage.     

Evaluation of a vermicompost and leachates on <i>Solidago x hybrida</i> and organic carbon mineralization under aerobic incubations

In the floriculture region of Tenancingo in the State of Mexico, the application of stabilized organic matter, such as vermicompost and leachates, contributes to improve the quality of the soil and plant nutrition. However, it is important to know the chemical composition of a vermicompost and the mineralization process. This is because the amount and speed of nutrient release which will be available to the crop will depend on that knowledge. The objective of the study was to evaluate the effect of the application of a vermicompost and leachates on various quantitative variables of Solidago x hybrid, and the mineralization of organic carbon under aerobic incubations. Vermicompost (74 and 36 g/kg soil), leachates (5 and 10 L/kg soil), 0.33 g/kg soil of chemical fertilizer Ca (NO₃)₂, and not treated soil (control) were applied under greenhouse conditions to evaluate their effects on plant growth variables. Mixtures of 100 g of soil with vermicompost and leachates were made in the laboratory which were incubated during 9 weeks to obtain the potentially mineralizable organic carbon (Corg PM) and the rate of mineralization (k) after adjusting an exponential model. In the greenhouse experiment there were no statistical differences after applying vermicompost and leachates on the quantitative variables (number of stems per plant, diameter of the panicle, fresh weight, plant length and stem diameter) with respect to the control (p>0.05). The effect among the applied doses was evident only for variables such as fresh weight, panicle length and stem diameter with respect to the control. In incubated soils, k values ranged between 0.209-0.325 C mg/kg soil/week. Only with the application of leachates in high doses two pools of organic matter were shown: one soluble labile (102.9 mg/kg soil) and the other hydrolysable (819 mg/kg soil). The soluble, labile fraction favored nutrient availability immediately after its application to the soil. However, a single pool of hydrolysable organic C (987-1074 mg/kg soil) was found when vermicompost was applied. It was associated with a release of organic matter during crop development, and to a possible stimulation of microbial activity. High values of electrical conductivity in vermicompost and leachates (8.2-11.7 mS/m) suggest a moderate application of both products.     

Integrated vermi-pisciculture--an alternative option for recycling of solid municipal waste in rural India

Vermicomposts as a biofertilizer can be a great option for pond manuring as they never cause any long term harm to the soil like chemical fertilizer. In this study vegetable and horticulture waste was used as an important media for vermiculture. Three separate cemented tanks (6 m(3) each) were used in the system as control tank, vermicompost fertilized tank and inorganic fertilizer manured tank. Monoculture of fish was carried out with cat fish, Clarias batrachus. The produced earthworms were used as fish feed. Regular monitoring of water parameter was conducted in three different ponds. Specifically, the algal biomass variation was quite helpful in analyzing the behavior of the ponds. NPK value of soil samples were analyzed intermittently to know the eutrophication level. Despite the hot summer temperature in northern part of India, which is not ideal for fish growth, we have recorded an encouraging growth performance in organic manured pond along with inorganic fertilizer treated and control pond. Among eutrophicated pond, the fish biomass from vermicompost fed pond showed an increasing trend compared to inorganic fertilizer treated pond. Water retention capacity of vermicompost pond soil was better in comparison to other ponds. Result shows that the low cost model by integrating two production system vermiculture and pisciculture could be a commercially and environmentally viable option.     

Side Effects of Neem Oil on the Midgut Endocrine Cells of the Green Lacewing Ceraeochrysa claveri (Navás) (Neuroptera: Chrysopidae)

We described the ultrastructure of Ceraeochrysa claveri (Navás) midgut endocrine cells in larva, pupa, and adult, and evaluated the side effects of ingested neem oil, a botanical insecticide obtained from the seeds of the neem tree (Azadirachta indica), on these cells. During the larval period, C. claveri were fed (ad libitum) Diatraea saccharalis (F.) eggs treated with neem oil at concentrations of 0.5%, 1%, or 2%. Transmission electron microscopy showed that two subtypes of endocrine cells, namely granular and vesicular, occurred in the midgut epithelium during the three stages of the life cycle. Both cell types did not reach the midgut lumen and were positioned basally in the epithelium. The endocrine cells did not show extensive infoldings of the basal plasma membrane, and there were numerous secretory granules in the basal region of the cytoplasm. In the granular endocrine cells, the granules were completely filled with a dense matrix. In the vesicular endocrine cells, the main secretory products consisted of haloed vesicles. Ultrastructural examination indicated that only the granular endocrine cells exhibited signs of morphologic changes of cell injury present in all life cycle stages after the larvae were chronically exposed to neem oil by ingestion. The major cellular damage consisted of dilatation and vesiculation of the rough endoplasmic reticulum and the development of smooth endoplasmic reticulum and mitochondrial swelling. Our data suggest that cytotoxic effects on midgut endocrine cells can contribute to a generalized disruption of the physiological processes in this organ due to a general alteration of endocrine function.