Behavioral and Physiological Effects of Neem (Azadirachta indica) Seed Kernel Extracts on Larval Parasitoid, Bracon hebetor

An aqueous suspension and an ethanolic extract of neem seed kernel (NSK) at 0.3, 0.6, 1.2, 2.5, and 5.0% concentrations were tested for ovipositional deterrency, feeding deterrency, toxicity, sterility, and insect growth regulatory effects on a larval parasitoid, Bracon hebetor. Neither NSK extracts delivered in the food or by contact influenced the B. hebetor oviposition (parasitization). They also did not cause parasitoid sterility through feeding, but they showed feeding deterrent effects for a limited period. Parasitoid eggs and pupae also were unaffected by the extracts tested. The parasitoid larvae, however, were killed by feeding contaminated host larvae and also through contact with neem extracts. Thus, a minimum safety period is suggested for inundative releases of B. hebetor in integrated pest management.     

Inhibition of Citrinin Production in Penicillium citrinum Cultures by Neem [Azadirachta indica A. Juss (Meliaceae)]

The efficacy of different concentrations of aqueous neem leaf extract (3.12 to 50 mg/mL) on growth and citrinin production in three isolates of Penicillium citrinum was investigated under laboratory conditions. Mycotoxin production by the isolates was suppressed, depending on the concentration of the plant extract added to culture media at the time of spore inoculation. Citrinin production in fungal mycelia grown for 21 days in culture media containing 3.12 mg/mL of the aqueous extract of neem leaf was inhibited by approximately 80% in three isolates of P. citrinum. High-performance liquid chromatography was performed to confirm the spectrophotometric results. Vegetative growth was assessed, but neem extract failed to inhibit it. Neem leaf extract showed inhibition of toxin production without retardation in fungal mycelia growth.     

Photooxidation of Nimbin and Salannin, Tetranortriterpenoids from the Neem Tree (Azadirachta indica)

Nimbin and salannin, major triterpenoids accompanying azadirachtin in extracts of neem (Azadirachta indica) seeds, were photooxidized by UV light in the presence of oxygen to more polar, unstable intermediates that rearranged on silica gel to two final products in which the furan ring had been oxidized to isomeric hydroxybutenolides. The isomeric hydroxybutenolides were also readily formed when a crude extract of triterpenoids from neem seeds was irradiated, and both isomers of salannin have been isolated from seeds. Photooxidation of nimbin and salannin proceeded much faster than that of azadirachtin. All photoproducts showed some biological activity againstSpodoptera littoralis, Locusta migratoria, andSchistocerca gregaria. Isonimbinolide was as potent as azadirachtin at inhibiting feeding in all three species, and it also inhibited the growth ofS. littoralis. Isosalanninolide showed potent antifeedant and growth inhibitory activity againstS. littoralis.     

Antifeedant activity of quassinoids

The antifeedant activity of 13 quassinoids of different structural types has been studied against the Mexican bean beetle (Epilachna varivestis Mulsant) 4th instar larvae and the southern armyworm (Spodoptera eridania Crawer) 5th instar larvae. All quassinoids tested displayed significant activity against the Mexican bean beetle and, thus, do not reveal a simple structure-activity relationship. Five quassinoids were active against the southern armyworm. Interestingly, four of these-bruceantin (I), glaucarubinone (VI), isobruceine A (VIII), and simalikalactone D (XI)-possess the required structural features for antineoplastic activity. The noncytotoxic quassin (X) is an exception; it is active against both pests.     

Host plant iridoid-based chemical defense of an aphid,Acyrthosiphon nipponicus,against ladybird beetles

A Rubiaceae-feeding aphid,Acyrthosiphon nipponicus, is seldom attacked by the ladybird beetle,Harmonia axyridis. A potent deterrent against the beetle was isolated from the aphid and identified as paederoside, an iridoid glycoside originating in the aphid's host,Paederia scandens. The iridoid content was as high as 2% of the intact body weight, and a large portion was found in the cornicle secretion.     

Effect of DIMBOA,an aphid resistance factor in wheat,on the aphid predatorEriopis connexa Germar (Coleoptera: Coccinellidae)

DIMBOA (2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one), a secondary metabolite found in cereal extracts, confers resistance in wheat to aphids. Its effect on beneficial organisms was tested on larvae of the aphid predatorEriopis connexa Germar. Larvae were fed until pupation on artificial diets to which different concentrations of DIMBOA (2–200g/g diet) were added, as well as on aphids that had been feeding on wheat seedlings with different DIMBOA levels (140–440 g/g fresh tissue). In diets, the effect of DIMBOA was greatest on survival of third-instar larvae and on the duration of the second and fourth instars. When aphids were provided as food, those that had fed on a wheat cultivar with an intermediate DIMBOA level led to a significantly longer larval duration in the predator than did those that fed on either low or high DIMBOA cultivars. Shortest predator development times were obtained with aphid prey that had fed on high DIMBOA seedlings. Higher DIMBOA levels in the plant appear to reduce aphid feeding rates (and rates of DIMBOA ingestion), decreasing aphid survival and minimizing the effect of the toxin on the predator.On leave from Departamento de Entomologia, Universidad Nacional Agraria La Molina, Apartado aéreo 456, Lima, Perú.     

Attraction and repulsion of the aphid,Cavariella aegopodii,by Plant Odors

Cavariella aegopodii is induced to land on traps by the monoterpene carvone, and the relevance of this to host-finding by the aphid is discussed. Catches are reduced by linalool. The interaction of plant chemicals in natural communities is discussed, and the possibility of using repellent chemicals for crop protection is suggested.     

Extraction of natural insecticide azadirachtin from neem (Azadirachta indica A. Juss) seed kernels using pressurized hot solvent

Plant-derived pesticides are gaining popularity on account of the increasing sustainable farming practices and also due to the environmental and health hazards resulting from excessive usage of their synthetic counterparts. The neem (Azadirachta indica A. Juss) tree, belonging to the Meliaceae family, is a storehouse of insecticidal azadirachtin, which possess antifeedant, growth disrupting and larvicidal properties against an array of agricultural insect pests. Pressurized Hot Solvent Extraction (PHSE) is fast emerging as an efficient means for recovering valuable active ingredients from natural plant matrices at an accelerated rate and with a reduced solvent consumption. The technique employing heated organic solvents at elevated pressures is a potential green substitute for conventional solvent extractions. The present study deals with the extraction of azadirachtin from defatted neem seed kernels (NSK) using pressurized methanol. Important operating variables like temperature, pressure, extraction time, solvent flow rate and particle size influencing the extraction efficiency were investigated in detail. The azadirachtin content in the extracts was determined by HPLC. The optimum conditions for maximizing azadirachtin yield were found to be 50 °C temperature, 50 bar pressure, −60 + 80 mesh particle size and 5 mL/min extractant flow rate. Under optimal conditions, 210.93 mg azadirachtin was extracted per 100 g defatted NSK within an extraction time of 100 min. Moreover, the proposed method resulted in 1.5 fold reduction in solvent consumption as compared to conventional maceration.     

Jensenone: Biological Reactivity of a Marsupial Antifeedant from Eucalyptus

The resistance of Eucalyptus to browsing mammals has been related to the level and type of formylated phloroglucinol compounds (FPCs) present in the leaf. The antifeedant activity of FPCs appears to depend on their aldehyde groups, but little else is known of their mode of action. We have sought to elucidate this further by examining the biological reactivity and disposition of jensenone, a model FPC. Neither jensenone nor any metabolites were detected in urine or feces of marsupial brushtail or ringtail possums that had ingested up to 725 mg x kg(-0.75). When jensenone was incubated in rat gastrointestinal segments in vitro, it rapidly disappeared. Jensenone also reacted rapidly with glutathione, cysteine, glycine, ethanolamine, and trypsin, and more slowly with acetylcysteine and albumin. Sideroxylonal, a more complex FPC, exhibited the same reactivity. Torquatone, a related compound that lacks both aldehyde groups and antifeedant activity, was unreactive. Mass spectroscopic analysis indicated that the adducts were Schiff bases formed between the aldehyde groups of FPCs and amine groups of the conjugating molecules. Successive adducts were formed with the two aldehyde groups of jensenone, and the four groups of sideroxylonal. The jensenone bis-glutathione adduct appeared to cyclize to the disulfide form. These findings suggest that the antifeedant effects of FPCs are due to their facile binding to amine groups on critical molecules in the gastrointestinal tract, leading to a loss of metabolic function. The consequent toxic reaction, probably involving chemical mediators such as 5-hydroxytryptamine (5HT), may cause colic, nausea, and a general malaise, resulting in anorexia.     

Winter host component reduces colonization by bird-cherry-oat aphid,Rhopalosiphum padi (L.) (homoptera,aphididae), and other aphids in cereal fields

Methyl salicylate, a volatile component ofPrunus padus, the winter host ofRhopalosiphum padi, was found to reduce colonization of the summer host by this aphid. The compound was identified by gas chromatographic analysis coupled with recordings from cells in the primary rhinarium on the sixth antennal segment of the aphid. Methyl salicylate eliminated the attractancy of oat leaves to spring migrants in olfactometer tests. In Sweden, this compound significantly decreased colonization of field grown cereals byR. padi and in the U.K., populations ofSitobion avenae andMetopolophium dirhodum were significantly lower on treated plots.     

Sex attractant pheromone of damson-hop aphid Phorodon humuli (Homoptera,aphididae)

Behavioral studies using an olfactometer demonstrated that sexual females (oviparae) of the damson-hop aphid,Phorodon humuli, release a pheromone to which males respond. Volatiles produced by the oviparae were analyzed by coupled gas chromatography-single cell recording from the secondary rhinaria on the male antenna and showed the presence of one peak with major activity. Coupled gas chrornatography-mass spectrometry suggested a nepetalactol, which was shown to have the 4aR, 7S, 7aS stereochemistry by synthesis from the corresponding nepetalactone isolated from the labiate plantNepeta mussinii. Although the stereochemistry at carbon-1 is not yet established, a synthetic sample comprising ca. 70% 1S and 30% 1R attracted highly significant numbers of males to water traps placed within and adjacent to a hop garden. Initial studies also indicated attraction of males in both the olfactometer and in the field by volatiles from the primary host.     

Activity of drimane antifeedants and related compounds against aphids,and comparative biological effects and chemical reactivity of (−)- and (+)-polygodial

A series of natural drimanes and related synthetic compounds was tested for antifeedant activity against aphids. Polygodial and warburganal were the most active. The synthetic compounds methyl 9-hydroxydrimenoate and 9-hydroxydrimenal, although active against lepidopteran larvae, were inactive against aphids. Natural (–)-polygodial and the synthetic (+) isomer showed similar levels of activity as aphid antifeedants and in phytotoxicity, fish toxicity, and human taste tests, but reacted at different rates with enantiomers of 1-phenylethylamine.     

Feeding deterrency of some 4-hydroxycoumarins and related compounds: Relationship to host-plant resistance of alfalfa towards pea aphid (Acyrthosiphon pisum)

A series of 3-acyl-4-hydroxycoumarins, structurally related to dicoumarol, as well as several alfalfa constituents including coumestrol were tested for their feeding deterrency towards the pea aphid. Feeding deterrency of the 3-acyl-4-hydroxycoumarins decreased as the size of the 3-acyl group increased.     

Plant-derived synergists of alarm pheromone from turnip aphid,Lipaphis (Hyadaphis) erysimi (Homoptera,Aphididae)

The turnip aphid,Lipaphis (Hyadaphis) erysimi, responds weakly to (E)--farnesene, the main component of the alarm pheromone, but the response is substantially increased by incorporating plant-derived isothiocyanates, identified in aphid volatiles by coupled gas chromatography-single-cell recording.     

Role of cuticular hydrocarbons of aphid parasitoids in their relationship to aphid-attending ants

Lysiphlebus cardui, the dominant aphidiid parasitoid of the black bean aphid,Aphis fabae cirsiiacanthoidis (Afc), on creeping thistle, is able to forage in ant-attended aphid colonies without being attacked by ants. Several behavioral observations and experimental studies led to the hypothesis thatL. cardui mimics the cuticular hydrocarbon profile of its host aphid. Chemical analysis of the cuticular extracts revealed that bothL. cardui and Afc exclusively possess saturated hydrocarbons:n-alkanes, monomethyl (MMA), dimethyl (DMA), and trimethyl alkanes (TMA). Comparison of the hydrocarbon profiles of parasitoid and aphid showed great qualitative resemblance between parasitoid and host:L. cardui possesses almost all host-specific compounds in addition to species-specific hydrocarbons of mainly higher molecular weight (>C₃₀). However, there is a lesser quantitative correspondence between parasitoid and host aphid. Furthermore, we analyzed the cuticular hydrocarbon profile of another parasitoid of Afc,Trioxys angelicae. This aphidiid species is vigorously attacked and finally killed by honeydewcollecting ants when encountered in aphid colonies. Its cuticular hydrocarbon profile is characterized by the presence of large amounts of (Z)-11-alkenes of chain lenghts C₂₇, C₂₉, C₃₁, and C₃₃, in addition to alkanes and presumably trienes. The role of the unsaturated hydrocarbons onT. angelicae as recognition cues for aphid-attending ants is discussed.     

Disruption of web structure and predatory behavior of a spider by plant-derived chemical defenses of an aposematic aphid

Two toxic and bitter-tasting cardenolides (cardiac-active steroids) were sequestered by the brightly colored oleander aphid,Aphis nerii B. de F., from the neotropical milkweed host plantAsclepias curassavica L. After feeding on milkweed-reared aphids, the orb-web spiderZygiella x-notata (Clerck) built severely disrupted webs and attacked fewer nontoxic, control aphids, whereas the webs of spiders fed only nontoxic aphids remained intact. The regularity and size of the prey-trapping area of webs were reduced significantly in proportion to the amount of toxic aphids eaten. The effects of toxic aphids on spider web structure were mimicked by feeding spiders the bitter-tasting cardenolide digitoxin, a cardenolide with similar steroidal structure and pharmacological activity to the two aphid cardenolides. These results show that the well-known effects of psychoactive drugs on spider web structure are more than interesting behavioral assays of drag activity. Similar effects, produced by plant-derived chemicals in the spider's aphid prey, are relevant to the ecology and evolution of interactions between prey defense and predator foraging.     

Sucrose esters of carboxylic acids in glandular trichomes ofSolanum berthaultii deter settling and probing by green peach aphid

Removal of type B trichome exudate fromSolanum berthaultii leaflets leads to a decrease in tarsal gumming and mortality and an increase in feeding by the green peach aphid,Myzus persicae. Type B trichome exudate of theS. berthaultii accession PI 473331 is composed of a complex of 3,3,4,6-tetra-O-acyl sucroses containing primarily short-chain branched carboxylic acids. The acyl constituents are primarily derived from 2-methylpropanoic, 2-methylbutyric, and 8-methylnonanoic acids but constituents derived fromn-decanoic and dodecanoic acids are also present. Sucrose esters inhibit settling and probing by aphids in glass feeding cages.     

Effect of Dimboa, a Hydroxamic Acid from Cereals, on Peroxisomal and Mitochondrial Enzymes from Aphids: Evidence for the Presence of Peroxisomes in Aphids

2,4-Dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), a hydroxamic acid involved in the resistance of cereals to aphids, was administered to adult individuals of the aphid Sitobion avenae in artificial diets. Effects on the cellular metabolism were inferred from the evaluation of several organelle marker enzymes. Catalase from peroxisomes and cytochrome c oxidase from mitochondria increased their activities about twofold when aphids were fed with 2 mM DIMBOA. The role of these enzymes in the metabolizing of xenobiotics by aphids is discussed. Biochemical and cytochemical evidences for the presence of peroxisomes in aphids are reported here for the first time.     

Antifeedant activity againstSpodoptera litera larvae and [13C]NMR spectral assignments of the meliatoxins

The tetranortriterpene, meliatoxin A₂, induced a significant level of antifeedant activity inSpodoptera litura F. larvae, while meliatoxin B₁, which lacks the ring D epoxide, was much less active. Complete [¹³C]NMR assignments for both compounds derived from homo- and heteronuclear short- and long-range couplings are given.     

Effects of Tansy Essential Oil from Tanacetum vulgare on Biology of Oblique-Banded Leafroller, Choristoneura rosaceana

The formulations TSW (tansy essential oil, surfactant, distilled water) and TSWD (tansy essential oil, surfactant, distilled water, dillapiol) increased the feeding rate of oblique-banded leafroller (OBLR) larvae with increasing concentrations of tansy essential oil. When tansy essential oil was continuously present at 0.1% and 1% in the diet, all OBLR larvae from susceptible and resistant populations died. The presence of 0.01% tansy essential oil in the diet affected female pupal weight, but not larval weight gain, larval developmental time and male pupal weight. Our results suggest that the concentration of tansy essential oil that significantly affects the developmental variables should be between 0.01% and 0.1% of tansy essential oil. Residues of the formulation TE (tansy essential oil, ethanol 95%) deterred oviposition of OBLR female.