Effects of azadirachtin on the nutrition and development of the tobacco budworm,Heliothis virescens (Fabr.) (Noctuidae)

The plant chemical azadirachtin was administered, either in artificial diet or by oral injection, to fifth instar larvae of the tobacco budworm, Heliothis virescens (Fabr.). At a dietary concentration of 0.03125 ppm, azadirachtin significantly reduced the amount of diet consumed and the weight gained by the larvae. Higher dietary concentrations (0.25 and 0.5 ppm) were necessary to reduce the efficiency of larval conversion of digested and ingested food, respectively. However, the approximate digestibility increased at the dietary concentration of 0.25 ppm.Orally injected azadirachtin (0.25 and 0.5 μg) delayed moulting to the pupal stage, produced defective pupae or adults, and inhibited development to the adult stage. Higher doses (5.0 and 10.0 μg) reduced the pre-pupal weight loss normally associated with pupation, and completely inhibited pupation. At the critical dose of 1.0 μg (the minimal dose that disrupted development to the pupal stage), azadirachtin had less of an effect on older than on younger larvae. Larvae injected on the first day of the fifth instar failed to pupate, whereas approx 40% of those injected on subsequent days pupated.The results suggest that azadirachtin affects H. virescens in a manner similar to other tested species of insects. The significance of these results, especially regarding hormonal events in the insects, is discussed.     

Role of the head in the ultrastructural midgut organization in Rhodnius prolixus larvae: evidence from head transplantation experiments and ecdysone therapy

Studies on the effects of decapitation, head transplantation and ecdysone therapy on the ultrastructural organization of the midgut in 5th-instar larvae of Rhodnius prolixus, were carried out. Control insects had a typical and significant organization of the epithelial cells (mainly microvilli, extracellular membrane layers and basal portion of the epithelial cells) of the midgut (stomach and intestine) during the entire period of the experiment. However, the host larvae, when decapitated 1 day after feeding, demonstrated significant changes in the ultrastructural organization of the epithelial cells of these compartments. In converse experiments, head transplantations from untreated donors 4-5 days after feeding into headless larvae sustained the ultrastructural organization of the epithelial cells in the midgut. Oral therapy with ecdysone (5 &mgr;g/mL of blood meal) in decapitated insects significantly reversed the altered organization of the stomach and intestine. These results point to a brain factor, possibly the prothoracicotropic hormone (PTTH) which stimulates ecdysteroid production in the prothoracic glands, may be a factor responsible, directly or indirectly, for the midgut cell organization in R. prolixus.     

Enhanced survival of shrimp, Penaeus (Marsupenaeus) japonicus from white spot syndrome disease after oral administration of recombinant VP28 expressed in Brevibacillus brevis

White spot syndrome virus (WSSV) disease is a major threat to shrimp culture worldwide. Here, we assessed the efficacy of the oral administration of purified recombinant VP28, an envelope protein of WSSV, expressed in a Gram-positive bacterium, Brevibacillus brevis, in providing protection in shrimp, Penaeus japonicus, upon challenge with WSSV. Juvenile shrimp (2-3g in body weight) fed with pellets containing purified recombinant VP28 (50mug/shrimp) for 2weeks showed significantly higher survival rates than control groups when challenged with the virus at 3days after the last day of feeding. However, when shrimp were challenged 2weeks after the last day of feeding, survival rates decreased (33.4% and 24.93%, respectively). Survival rate was dose-dependent, increasing from 60.7 to 80.3% as the dose increased from 1 to 50mug/shrimp. At a dose of 50mug/shrimp, the recombinant protein provided protection as soon as 1day after feeding (72.5% survival). Similar results were obtained with larger-sized shrimp. These results show that recombinant VP28 expressed in a Gram-positive bacterium is a potential oral vaccine against WSSV.     

Influence of brain and azadirachtin on Trypanosoma cruzi development in the vector, Rhodnius prolixus.

Studies on the effects of decapitation, head transplantation, azadirachtin, and ecdysone therapy on the ultrastructural organization of the midgut of Rhodnius prolixus, a vector of the protozoan Trypanosoma cruzi, show a distinct effect on the organization of the epithelial cells. When insects are decapitated or treated with azadirachtin, the ultrastructural organiza tion of these compartments changed significantly and drastically blocked the development of T. cruzi infection. In converse experiments, head transplantation or oral therapy with ecdysone significantly re versed the T. cruzi infectivity and reestablished the organization of the stomach and intestine in decapitated or azadirachtin-treated insects. These results indicat that a brain factor, possibly the prothoracicotropic hormone which stimulates ecdysteroid production on the prothoracic glands, may act directly or indirectly on both the midgut cell organiza tion and the intestinal microenvironment, interfering in the trypanosome survival and infection of the vector R. prolixus.     

Effect of blood components, abdominal distension, and ecdysone therapy on the ultrastructural organization of posterior midgut epithelial cells and perimicrovillar membranes in Rhodnius prolixus

The effects of blood components, nerve-cord severance, and ecdysone therapy on the posterior midgut epithelial cells of 5th-instar Rhodnius prolixus nymphs 10 days after feeding were analyzed by transmission electron microscopy. Cutting the nerve-cord of the blood-fed insects partially reduced the development of microvilli and perimicrovillar membranes (PMM), and produced large vacuoles and small electrondense granules; insects fed on Ringer's saline diet exhibited well developed microvilli and low PMM production; swolled rough endoplasmatic reticulum and electrondense granules; Ringer's saline meal with ecdysone led to PMM development, glycogen particles, and several mitochondria in the cytoplasm; epithelial cells of the insects fed on Ringer's saline meal whose nerve-cord was severed showed heterogeneously distributed microvilli with reduced PMM production and a great quantity of mitochondria and glycogen in the cytoplasm; well developed microvilli and PMM were observed in nerve-cord severed insects fed on Ringer's saline meal with ecdysone; Ringer's saline diet containing hemoglobin recovered the release of PMM; and insects fed on human plasma showed slightly reduced PMM production, although the addition of ecdysone in the plasma led to a normal midgut ultrastructural organization. We suggest that the full development of microvilli and PMM in the epithelial cells depends on the abdominal distension in addition to ingestion of hemoglobin, and the release of ecdysone.     

Cellular immune response in Rhodnius prolixus: role of ecdysone in hemocyte phagocytosis

In this paper we investigate in vivo and in vitro effects of orally administered azadirachtin and ecdysone on the phagocytic responses of Rhodnius prolixus 5th-instar larval hemocytes to the yeast Saccharomyces cerevisiae. Groups of insects fed non-treated blood (control) and insects that received azadirachtin plus ecdysone in the blood meal were inoculated with yeast cells in the hemocele. The injected yeast cells disappeared rapidly from the hemolymph, being removed completely by 90min after inoculation. In the insects treated only with azadirachtin the clearance of free yeast circulating particles was significantly delayed compared to the two previously mentioned groups. It was demonstrated that the binding of yeast cells to hemocytes was reduced in the insects treated only with azadirachtin in comparison to both non-treated control and azadirachtin plus ecdysone-treated groups. Phagocytosis occurred when yeast cells were added to hemocyte monolayers prepared with hemolymph from blood fed insects, treated or not with azadirachtin plus ecdysone, so that yeast cells were rapidly bound to hemocytes and internalized in high numbers. By contrast, insects treated with azadirachtin exhibited a drastic reduction in the quantity of yeast cell-hemocyte binding and subsequent internalization. In all groups, the hemocytes attached to the glass slides were predominantly plasmatocytes. The magnitude and speed of the cellular response suggests that hemocyte phagocytosis is one of the main driving forces for the clearance of free circulating yeast cells from the hemolymph. We propose that ecdysone modulates phagocytosis in R. prolixus larvae, and that this effect is antagonized by azadirachtin.     

Differential cytotoxic effects of azadirachtin on Spodoptera frugiperda and mouse cultured cells

Azadirachtin is a natural biopesticide extracted from the neem tree (Azadirachta indica); however, its safety to mammals is not well documented. This study examined the influence of azadirachtin on the proliferation and morphology of Spodoptera frugiperda insect cells (SF-9), and mouse erythroleukemia cells (MEL-GM 86). Cell kinetic studies conducted at 24 h intervals for 144 h showed that azadirachtin treatments (0.05, 0.20 and 0.50 g/ml) inhibited cell multiplication of the SF-9 cells. This inhibitory effect was dose dependent. Conversely, azadirachtin at 30 g/ml did not reduce MEL-GM 86 cell proliferation. SF- 9 cells displayed an increase in cell swelling and cell abnormalities after 48 h of treatment with azadirachtin, whereas no such changes were detected in MEL-GM 86 cells. A scanning electron microscope study revealed that treatment with an azadirachtin concentration as low as 0.10 g/ml induced cell swelling and caused distortions on the cell surface ultrastructure of SF-9 cells characterized by blebbing and holes after 48 h of treatment. However, these effects were not seen in the azadirachtin-treated MEL- GM 86 cells.     

Accumulation of cadmium by the fourth instar larva of the fly Chironomus thummi

Accumulation and effects of cadmium were investigated in Chironomus thummi larvae exposed to 10, 100 and 250 mug radiolabeled Cd/1 for up to 4 days. (1) After 4 days, average cadmium accumulation was 6.6 ng Cd/mg dry weight (10 mug Cd/1 exposure) and 177 ng Cd/mg dry weight (250 mug Cd/1 exposure). (2) Dissection studies showed that by 32 hr of exposure to both cadmium concentrations, 63.5-81.4% of accumulated cadmium was confined to the posterior midgut epithelium. Light microscope autoradiography similarly showed accumulations of cadmium in posterior midgut epithelium and smaller amounts in fat body and muscle. Little cadmium was associated with Malphigian tubules, haemocoel, anterior midgut or exoskeleton. (3) After exposure to 10 or 250 mug Cd/1, 60-75% of cadmium in ultracentrifuged homogenates of whole animals or dissected guts was associated with the resulting supernatant. When supernatants were further analyzed by gel chromatography, cadmium eluted with both a high and low molecular weight peak. The relative proportions of cadmium in the two peaks varied with concentration and length of exposure. (4) Transmission electron microscopy of posterior midgut cells from animals exposed to cadmium demonstrated frequent mitochondrial lesions. Exposure to high cadmium concentrations caused some posterior midgut cells to undergo generalized structural degeneration.     

Azadirachtin affects growth and endocrine events in larvae of the tobacco hornworm, Manduca sexta

Injection of azadirachtin in freshly emerged last-instar larvae of Manduca sexta elicited different reactions according to the dose administered. At low doses, pupation occurred in most of the cases, but the resulting pupae were defective for the most part. Individuals treated with higher doses usually did not fully complete development, moulting to supernumerary larvae or dying as larvae (sometimes at the wandering stage) after varying periods of survival. The haemolymph ecdysteroid titre of individuals treated with 2 μg azadirachtin/g bodyweight showed characteristic changes which are presumed to cause the disorders in the last stages that normally lead to pupation. Injection of moulting hormone in azadirachtin-treated individuals at certain times during the penultimate stage elicited no reduction of the azadirachtin-induced effects. It is shown that azadirachtin is able to inhibit development even when individuals performed a complete moult after the treatment.     

Laboratory evaluation of the toxicity of systemic insecticides for Control of Anoplophora glabripennis and Plectrodera scalator (Coleoptera: Cerambycidae)

Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae) is one of the most serious nonnative invasive forest insects discovered in North America in recent years. A. glabripennis is regulated by federal quarantines in the United States and Canada and is the subject of eradication programs that involve locating, cutting, and chipping all infested trees. Other control methods are needed to aid in eradication and to form an integrated management program in the event eradication fails. We conducted laboratory bioassays to determine the toxicity of two systemic insecticides, azadirachtin and imidacloprid, for potential control of A. glabripennis and the cottonwood borer, Plectrodera scalator (F.) (Coleoptera: Cerambycidae), a closely related native cerambycid. Larvae of both cerambycid species were fed artificial diet with dilutions of azadirachtin or imidacloprid for 14 wk. Both insecticides exhibited strong antifeedant effects and some toxicity against A. glabripennis and P. scalator larvae. For A. glabripennis, the highest larval mortality at the end of the bioassay was 60% for larvae fed artificial diet treated with azadirachtin (50 ppm) or imidacloprid (1.6 ppm). For P. scalator, the highest larval mortality at the end of the bioassay was 100% for larvae fed artificial diet treated with azadirachtin (50 ppm) or imidacloprid (160 ppm). At 14 wk, the LC50 values for P. scalator were 1.58 and 1.78 ppm for azadirachtin and imidacloprid, respectively. Larvae of both species gained weight when fed diet treated with formulation blanks (inert ingredients) or the water control but lost weight when fed diet treated with increasing concentrations of either azadirachtin or imidacloprid. In a separate experiment, A. glabripennis adults were fed maple twigs treated with high and low concentrations of imidacloprid. A. glabripennis adult mortality reached 100% after 13 d on twigs treated with 150 ppm imidacloprid and after 20 d on twigs treated with 15 ppm imidacloprid. There was no visible feeding by A. glabripennis adults on twigs treated at the higher imidacloprid rate, and feeding was significantly reduced for adults placed on twigs treated at the low imidacloprid rate compared with adults on untreated twigs. In summary, imidacloprid and azadirachtin had both antifeedant and toxic effects against A. glabripennis and P. scalator and have potential for use in management programs. Based on our results, the delivery of high and sustained insecticide concentrations will be needed to overcome the antifeedant effects and lengthy lethal time for both larvae and adults exposed to these insecticides.     

Phenol oxidases from Rhodnius prolixus: Temporal and tissue expression pattern and regulation by ecdysone

Rhodnius prolixus 5th instar nymphs have significant PO enzymatic activity in the anterior midgut, fat body and hemolymph. The tissue with the major amount of PO activity is the anterior midgut while those with higher specific activities are the fat body and hemolymph. In this work the temporal pattern of PO enzymatic activity in different tissues was investigated. In fat body, PO peaks occur at 7, 12 and 16 days after a blood meal. In hemolymph, PO diminishes until day 7, and then recovers by day 14. In the anterior midgut tissue, PO peaks on day 9 and just before ecdysis; a similar pattern was observed in the anterior midgut contents. Some of these activities are dependent on the release of ecdysone, as feeding blood meal containing azadirachtin suppresses them and ecdysone treatment counteracts this effect. These results suggest that during the development of the 5th instar, the insect has natural regulating cycles of basal PO expression and activation, which could be related to the occurrence of natural infections. The differences in temporal patterns of activity and the effects of azadirachtin and ecdysone in each organ suggest that, at least in R. prolixus, different tissues are expressing different PO genes.     

Modification of the nutritional parameters and of midgut biochemical and absorptive functions induced by the IGR fenoxycarb in Bombyx mori larvae

Fifth instar larvae of B. Mori were topically or orally treated with increasing amounts of the Insect Growth Regulator (IGR) fenoxycarb in a single application, in order to determine its effects on the nutritional parameters, the midgut functional activities and the growth of the silk glands. The IGR affected in a dose-dependent manner the progress of the life cycle of the insect, causing a delay or inhibition of spinning, alteration of the feeding behaviour, decrease of the nutritional parameters, impairment of the growth of the silk glands, and an increased mortality during larval-pupal transformation. Measurement of leucine uptake into midgut brush border membrane vesicles and midgut histochemistry revealed a reduced absorption of leucine by the midgut and a large alteration of a number of midgut enzyme activities as a result of treatments with a high dose of fenoxycarb (2.5 μg). Treatments with a dose of 2.5 femto g/larva caused an increase in leucine uptake by the midgut, an increased weight of the cocoon shell, and a modification of some midgut enzyme activities. The lepidopteran midgut appears to be a larval organ that responds promptly to the exposure to fenoxycarb. The epithelial columnar cells modify their absorptive functions, at least with regard to amino acid uptake, as well as their metabolic activity, with a modification of the oxidative status of the cells that is detectable with a single dose of the chemical as low as few fg/larva. Arch. Insect Biochem. Physiol. 39:18–35, 1998. © 1998 Wiley-Liss, Inc.     

Cytotoxicity of azadirachtin A in human glioblastoma cell lines

The neem toxin azadirachtin A exhibits selective toxicity on insects. Despite its well-proven efficacy, the mode of action of this toxin remains obscure. The toxicity on vertebrate cells compared to insect cells is also not well characterized. We have cultivated six human glioblastoma cell lines G-28, G-112, G-60 (TP53 mutant) and G-44, G-62, G-120 (TP53 wild-type) in the presence of 28 microM of azadirachtin. This toxin concentration was chosen because it represents the 25 to 50% lethal dose in the glioma cells. Toxicity was measured in terms of cell proliferation (binucleation index), formation of micronuclei and cell survival. In the TP53 mutant cell lines, azadirachtin reduced the proportion of dividing cells and induced formation of micronuclei. Except for G-44 which showed a decrease in binucleation index, proliferation in the TP53 wild-type cell lines was unaffected by azadirachtin. In the TP53 wild-type cell lines, the decrease in micronuclei frequency is attributed to fewer cells entering mitosis to produce micronuclei. This is also apparent from the low surviving fractions. Cell survival was suppressed by 25-69% in all cell lines. The reduction of cell survival is a clear indication that azadirachtin affects reproductive integrity and cell division. The induction of micronuclei reflects DNA damage. Similar studies on damage induction in insect cell lines could elucidate the processes which precede the antifeedant and antimoulting effects of azadirachtin and other neem toxins in insects.     

Impact of azadirachtin, an insecticidal allelochemical from neem on soil microflora, enzyme and respiratory activities

The effect of 10% azadirachtin granules (alcoholic extract of neem seed kernel mixed with China clay) was studied on the population of bacteria, actinomycetes, fungi, Azotobacter and nitrifying bacteria; soil dehydrogenase, phosphatase and respiratory activities on 0, 15th, 30th, 60th and 90th days after application in sandy loam soil collected from the fields. It was observed that baring the Azotobacter sp., azadirachtin at all the doses exerted a suppressive effect on the rest of the microbial communities and enzyme activities in the initial 15 day period. The population of bacteria, actinomycetes besides phosphatase and respiratory activities recovered after 60th day and subsequently increased significantly. The fungi and nitrifiers were most sensitive groups as their numbers were reduced significantly throughout the studies. The two times and five times recommended dose of azadirachtin had very high biocidal effects on the soil microorganisms and its activities. However, analysis of the data by the Shannon Weaver index showed that azadirachtin reduces both the form and functional microbial diversity at all doses.     

Effects of azadirachtin and of simpler epoxy-alcohols on survival and behaviour of Galleria mellonella (Lepidoptera)

Abstract:  Investigations into the toxicity of three simpler molecules based on the epoxy-alcohol fragment of azadirachtin have revealed insecticidal activity on the greater wax moth Galleria mellonella L. larvae. The simpler epoxy-alcohols doses giving 50% mortalities (LD₅₀) for G. mellonella larvae were in the increasing order from glycidol (0.022 mg/g), 4,5-epoxy-2-pentanol (0.068 mg/g) and finally, glycerol diglycidyl ether (0.147 mg/g). The three epoxy-alcohols exhibited higher insecticidal activity when compared with the commercial neem product for which the dose giving 50% mortalities was 10.6 mg/g and to azadirachtin that killed the larvae only by injection (dose of 0.20 mg/g of larvae body weight). Our results confirm the importance of the epoxy-alcohol junction between the two parts of the azadirachtin molecule for the biological activity. Other effects of the epoxy-alcohols tested were blackening of larvae and morphological deformities of some adults hatching. In future, the molecules should be complexified (degree of ramification, length of chain and presence of bulky ramified substituent) to obtain an insecticide as toxic for insects only and environmentally safe as azadirachtin but more stable, and their physiological activities on insect's tissues and cells should be studied.     

Systemic antifeedant effects of azadirachtin on the peach-potato aphidMyzus persicae

The electrical penetration graph (EPG) method was used to analyse the feeding behaviour of apterous, adultMyzus persicae (Sulzer) (Homoptera: Aphididae) onNicotiana clevelandii (Gray) seedlings, treated systemically with azadirachtin. A preliminary experiment showed that the effects of tethering aphids for EPG recording were minimal. The percentage of the 9 h recording period devoted to non-penetration activities, and to stylet pathway patterns increased as the azadirachtin concentration in the root treatment increased. The number of probes initiated, and the numbers of sieve tube penetrations also increased with increased azadirachtin concentration. The mean time elapsing between the initiation of the first probe to reach a sieve element and contact with this tissue was not significantly altered by azadirachtin treatment. However, azadirachtin treatment significantly reduced the percentage of probes that reached sieve elements and increased non-penetration activity before and after the first perid of ingestion from the sieve elements. The percentage of the recording period spent in the EPG pattern associated with sieve tube penetration was significantly reduced by an azadirachtin concentration of 300 ppm, and the duration of each individual penetration was significantly reduced by an azadirachtin concentration of 100 ppm. When the total EPG was split into 3 h periods, significant interactions were seen between time period and azadirachtin concentration for the duration of non-penetration, pathway, and sieve tube penetration patterns.     

几种植物源化合物对棉铃虫海藻糖酶活性及相关碳水化合物含量的影响

碳水化合物对昆虫的能量代谢和物质合成具有重要的作用。本研究选用2种一般性生物碱（氢溴酸东莨菪碱和烟碱）以及2种β-葡萄糖苷类化合物（七叶灵和皂角苷）, 研究其在不同浓度下对棉铃虫Helicoverpa armigera (Hübner)幼虫体内海藻糖酶活性及相关碳水化合物代谢的影响。结果表明: 用饲喂法处理3龄幼虫96 h后, 皂角苷对棉铃虫幼虫的活体抑制效果明显, 且随添加物浓度增高, 棉铃虫死亡率上升, 10, 20, 40 g/L浓度下棉铃虫的均重分别是0.194, 0.089和0.034 g, 分别为对照的86.99%, 39.91%和15.24%。对海藻糖酶活性及其相关代谢酶的测定结果表明, 2种苷类化合物显著抑制中肠海藻糖酶活性, 饲喂40 g/L皂角苷的试虫中肠海藻糖酶比活力仅是对照组的54.21%; 饲喂30 g/L七叶灵的试虫中肠海藻糖酶比活力为对照组的83.73%。而2种生物碱类化合物显著抑制血淋巴和脂肪体中海藻糖酶活性, 20 g/L氢溴酸东莨菪碱对棉铃虫血淋巴和脂肪体组织的海藻糖酶活性抑制率分别为7.24%和71.43%; 而20 g/L烟碱对试虫血淋巴和脂肪体组织的海藻糖酶活性抑制率为26.29%和33.44%。用氢溴酸东莨菪碱、 烟碱和七叶灵处理试虫后, 血淋巴海藻糖含量都有所增高。4种化合物能够导致试虫糖原磷酸化酶活性变化, 其中, 皂角苷在中肠和脂肪体表现为显著抑制作用, 而随外源化合物浓度变化, 糖原含量和糖原磷酸化酶活性表现为此消彼长关系。饲喂4种植物源化合物的试虫血淋巴中葡萄糖浓度变化和其海藻糖变化一致。本研究证明β-葡萄糖苷类化合物是海藻糖酶抑制剂, 在作为先导化合物进行农药创制开发方面具有重要意义。     

Ultrastructural changes of the midgut epithelial cells in feeding and moulting nymphs of the tick Haemaphysalis longicornis

The midgut epithelial cells in nymphs fed on laboratory rabbits were examined during feeding and after detachment. The midgut epithelium at the unfed stage consisted of digestive cells of lower activity, containing such nutritive substances as protein, lipid and glycogen. As feeding proceeded, the cells became active in intracellular digestion. At the middle of the feeding stage, the spent digestive cells derived from the active digestive cells began to be replaced by the new digestive cells of lower activity. After detachment, the pinocytotic activity of the above cells increased greatly, and the digestive activity increased to some extent. As a result, many large endosomes were formed by fusion of numerous pinosomes. Thereafter, endosomes decreased in size as digestion proceeded and there was an increase of haematin granules. On day 7 after detachment, the new digestive cells of lower activity, belonging to the 'nutritional reserve' type, appeared adjacent to the spent digestive cells which had almost exhausted all endosomes, and these new cells had completely replaced the spent cells by day 3 after moulting.     

[Ciliogenesis in the mucous cells of the quail oviduct. II. Hormonal control]

The hormonal control of ciliogenesis and transformation of mucous cells was studied in the oviduct (magnum) of ovariectomized quails. Estradiol benzoate induces ciliogenesis with doses varying from 10 mug/day to 100 mug/day after 6 days of treatment. With 100 mug/day, differentiation of some mucous cells is also induced as well as the formation of transitory "mixed cells" which are in the process of ciliogenesis and contain mucous granules. Associated with progesterone (1 mg/day), estradiol benzoate (10 mug/day) induces the differentiation of mucous cells and ciliated cells. The luminal epithelium of quails injected with this mixture is similar to the luminal epithelium observed in the oviduct of laying quails. With the same dose of progesterone (1 mg/day) and 20 mug/day of estradiol benzoate for 6 days, ciliogenesis is completely inhibited. All epithelial cells are secretory cells. Transformation of 50% of the mucous cells into ciliated cells is obtained by following the previous estradiol-progesterone treatment with the injection of estradiol benzoate (20 mug/day) for 3 days. Divisions of mucous cells were also observed. It is also possible to induce ciliogenesis in some mucous cells by withdrawing both hormones for 3 days. In this case, no cell divisions were observed.     

The effect of neem allelochemicals on nutritional physiology of larval Spodoptera litura

Neem allelochemicals azadirachtin, salannin, nimbinene and nimbin were administered to different larval instars of the tobacco armyworm, Spodoptera litura orally in artificial diet, topically or via injection. Nutritional analyses revealed strong antifeedant and growth regulatory effects of azadirachtin which were independent of each other. While salannin and nimbinene induced concentration dependent feeding deterrence only; nimbin was inactive to the 1000 ppm level against this insect species. One of the causes of the reduced growth rate of azadirachtin treated insects was due to an increase in the costs associated with growth. This was relative to a drastic reduction in the activity of gut trypsin. Salannin and nimbinene, however, did not interfere with the trypsin activity of the gut. These results and those from nutritional studies suggest that salannin and nimbinene have no toxicity mediated effects on S. litura larvae and antifeedant activity is a result of the effects on deterrent and other chemoreceptors. The fact that azadirachtin directly or indirectly inhibits the secretion of trypsin by the enzyme-secreting cells of the gut is discussed.