Deterrence coding by a larval Manduca chemosensory neurone mediating rejection of a non-host plant, Canna generalis L.

Abstract. The physiological basis of phagodeterrence was studied electrophy-siologically and behaviourally in the phytophagous caterpillars Manduca sexta and Manduca quinquemaculata. The model unacceptable non-host plant was the canna lily, Canna generalis. A strongly deterrent extract was obtained from fresh leaves of canna by extraction with hot ethanol or ethyl acetate in a blender. Behavioural rejection of these extracts was similar to that of fresh leaves, although less intense. In contrast, blender extracts using other solvents, as well as leaf surface rinses, were phagostimulant or neutral. Chromatographic fractionation of the deterrent ethanolic extract showed the active principles to be moderately polar and separable into two fractions. Previous ablation experiments had shown that the medial maxillary styloconica and epipharyngeal sensilla are the two most important chemosensory organs in mediating behavioural rejection of canna leaves; if only one of these organs is spared, the animal completely rejects canna. We investigated the neural responses of the medial styloconica and their contribution to the sensory coding responsible for this phagodeterrence. The active fractions of the deterrent ethanolic extract elicited a vigorous response from one chemosensory neurone in the medial styloconica. This neurone is distinguishable from others in the medial styloconica by its unique temporal response parameters and the characteristic shape changes of its action potentials. The response frequency of this neurone correlates with the degree of phagodeterrence in a dose-dependent manner. Threshold deterrence occurs at a concentration of extract (1%) that elicits firing in this neurone at a rate of c. 50 spikes/s peak instantaneous frequency and 30 total spikes in the first Is. We conclude that this is a ’deterrent neurone’ in the sense that vigorous response from this neurone is a sufficient sensory code for behavioural rejection of canna. Thus input from a single sensory neurone is capable of blocking feeding, since only one (unilateral) medial styloconicum is needed to mediate this rejection.     

Habituation in Frankliniella occidentalis to deterrent plant compounds and their blends

Feeding and oviposition deterrence of three secondary plant compounds and their 1:1 blends to adult female Frankliniella occidentalis Pergande (Thysanoptera: Thripidae) and the potential for habituation of the thrips to the pure compounds and the 1:1 blends at various concentrations were investigated. In choice assays, we tested dose‐dependent feeding and oviposition deterrence of the two fatty acid derivatives methyl jasmonate and cis‐jasmone, the phenylpropanoid allylanisole, and their blends when directly applied to bean leaf discs. The concentration required to reduce the feeding damage by 50% relative to the control treatment (FDC₅₀) was lowest for cis‐jasmone and highest for allylanisole. The feeding deterrent effect of both jasmonates was increased when blended with allylanisole. Feeding deterrence and oviposition deterrence were strongly correlated. In no‐choice assays conducted over four consecutive days, we discovered that dilutions at low concentrations (FDC₁₅) applied to bean leaves resulted in habituation to the deterrents, whereas no habituation occurred at higher concentrations (FDC₅₀). We observed a tendency that the 1:1 blends reduce the probability that thrips habituate to the deterrent compounds. Our results may be useful in the development of integrated crop protection strategies with the implementation of allelochemicals as pest behaviour‐modifying agents.     

Gustatory perception of phytoecdysteroids in Plodia interpunctella larvae

Phytoecdysteroids are steroidal compounds produced by various plants that disrupt growth and development of insects eating them. They exhibit an insecticidal activity on a number of insect pests such as Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae). In this study, we further evaluated whether phytoecdysteroids deter larvae of this species from feeding, by using four phytoecdysteroid molecules, commonly occurring in plants: 20‐hydroxyecdysone (20E), ponasterone A (PonA), polypodine B (PolB), and makisterone A (MakA). Fourth instar P. interpunctella avoided contact with food pellets treated with these phytoecdysteroids in a dose‐dependent way (2–30 mm ). In order to test whether this avoidance was mediated by taste sensitivity, we recorded the responses of taste neurons located in the lateral and medial sensilla styloconica of the galea. At least one neuron responded to each of these compounds in both sensilla. The neuron located in the medial sensillum had a detection threshold of 10^?6 m for PonA, 10^?4 m for 20E and PolB, and 10^?3 m for MakA. The lateral sensillum neuron responded with less intensity and its detection threshold was 10 times higher than that of the medial sensillum neuron. These results indicate that phytoecdysteroids are detected as deterrent stimuli by P. interpunctella larvae and that small structural differences significantly affect their biological activity.     

Antennal sugar sensitivity in the click beetle Agriotes obscurus

The occurrence of salt‐, sugar‐sensitive neurones and a mechanoreceptor neurone in the antennal hair‐like gustatory sensilla of the click beetle Agriotes obscurus L. (Coleoptera, Elateridae) is demonstrated using the electrophysiological sensillum tip‐recording technique. The stimulating effect of 13 water soluble sugars at 100 mm is tested on the neurones of these sensilla. Sucrose and fructose are the two most stimulating sugars for the sugar‐sensitive neurone, evoking almost 30 spikes s^?1 at 100 mm . The stimulating effect of arabinose, glucose, mannose, maltose and raffinose is three‐ to five‐fold lower, in the range 5.9–9.6 spikes s^?1. The remaining six sugars, xylose, galactose, rhamnose, cellobiose, trehalose and lactose, have very low (<1 spikes s^?1) or no ability to stimulate the sugar‐sensitive neurone. Concentration/response curves of the sugar‐sensitive neurone to sucrose, fructose and glucose at 0.01–100 mm overlap to a large extent in hibernating, cold reactivated and reproductively‐active beetles. A remarkable 9–50% decrease in the number of spikes evoked by 100 mm fructose and 10–100 mm sucrose occurs, however, in reproductively‐active beetles in June compared with beetles at the beginning of hibernation in October. These findings show that A. obscurus is capable of sensing a wide range sugars via their antennal gustatory sensilla.     

Electrophysiological responses of the chemoreceptor neurones in the antennal taste sensilla to plant alkaloids and glucosides in a granivorous ground beetle

The electrophysiological response of chemoreceptor neurones from the antennal chaetoid taste sensilla of the omnivorous ground beetle Pterostichus oblongopunctatus to several plant alkaloids and glucosides is investigated. A quinine‐sensitive neurone responding to quinine and quinine hydrochloride is found, most probably related to the granivorous feeding habit of P. oblongopunctatus. The response to quinine hydrochloride is concentration‐dependent at 0.001–50 mm , with the response threshold at 0.01 mm and a maximum rate of firing of 67 spikes/s at 50 mm . The stimulatory effect of caffeine is very weak, where the firing rate increases by only 1.4 spikes/s at a concentration of 10 mm compared with that evoked by a control stimulus. In addition, both quinine and quinine hydrochloride strongly inhibit spike production by the salt‐ and pH‐sensitive neurones when presented in mixtures with 10 mm NaCl. Several tested plant secondary compounds (i.e. salicin, sinigrin, caffeine and nicotine), which have only little or no effect on the firing rate of the quinine‐sensitive neurone, greatly reduce the responses of the salt‐ and pH‐sensitive neurones. The results of the present study suggest that the antennal taste sensilla of P. oblongopunctatus may detect plant defensive compounds both through the activation of a quinine‐sensitive neurone and via peripheral inhibition of other chemoreceptor neurones of the taste sensillum.     

Oviposition and feeding behavior of the maize borer,Chilo partellus,in response to eight essential oil allelochemicals

Eight essential oil compounds were evaluated against the maize borer, Chilo partellus (Swinhoe) (Lepidoptera: Pyralidae), to determine their influence on oviposition and feeding behavior. Thymol was the most active compound against C. partellus when evaluated as an oviposition deterrent (ODI₅₀ = 1.36 mg ml^?1), an ovicide (LC₅₀ = 2.06 mg ml^?1), or a feeding deterrent (FI₅₀ = 141.8 μg cm^?2) in laboratory experiments. The results corroborated with greenhouse experiments, in which egg laying on maize plants was inhibited significantly when the treatment was 15 times the concentration used in the laboratory experiments. However, there was neither any correlation between oviposition deterrence and feeding inhibition, nor between oviposition deterrence and ovicidal action. Apparently, toxicity per se or ovicidal action does not play any role in choice of oviposition in C. partellus. This was more obvious when the efficacy of compounds was compared in greenhouse experiments where ODI values and number of eggs laid on the treated leaves did not differ significantly in choice and no‐choice situations. Apparently, toxicity plays an important role in predicting host plant choice, but behavioral response in terms of oviposition preference is independent of toxic action, particularly for non‐host toxins. Thus, the same compounds affecting oviposition behavior on the one hand and having ovicidal or feeding deterrent properties on the other could be useful in field situations in any area‐wide integrated pest management model.     

EFFECT OF FEEDING STIMULANT ON THE FEEDING BEHAVIOR AND MORTALITY OF HELICOVERPA ARMIGERA ON DIETS WITH BACILLUS THURINGIENSIS*

Abstract Bacillus thuringiensis (Bt) was noticed to be highly deterrent to the feeding of Helicoverpa armigera larva when incorporated into diets, even at the non‐lethal dosage. As the dosage of Bt increased, the deterrence showed a tendency of elevation. Feeding stimulant, FS‐1 could attract larvae for feeding, and increase larval feeding time and consumption. Addition of 1% (vlv) FS1 could alleviate the deterrent effect of Bt, especialy in low concentration. In no‐choice test, it was observed that while 1% (v/v) FS1 was added, LC₅₀, of Bt underwent a 41% decrease, but in the choice test, the mortality of larvae was not significantly enhanced.     

Effects of pyrrolizidine alkaloids and sesquiterpenes on snail feeding

Summary We determined in the laboratory the feeding response of two populations of the generalist herbivorous snail Arianta arbustorum (Helicidae) towards the composite Adenostyles alliariae and towards various allelochemicals. These were: a pyrrolizidine alkaloid (PA) extract of Adenostyles leaves; senecionine (a PA present in Adenostyles); retrorsine (a PA not present in Adenostyles) and two sesquiterpene (ST) fractions from Adenostyles: a mixture of the STs adenostylone and neoadenostylone, and deacyladenostylone. Tertiary PAs and PA N-oxides were tested separately. For each allelochemical, we tested whether it was deterrent or whether it induced changes of feeding behaviour (i.e. whether it had pre- or postingestive effects), and whether the effects were more pronounced with younger (smaller) snails. The tertiary PA extract from Adenostyles was deterrent, especially for young snails, but did not induce changes of feeding behaviour. Tertiary PA senecionine was deterrent for young snails only and induced changes of feeding behaviour. Also, consumption of untreated Petasites was higher after this treatment. Tertiary PA retrorsine was not deterrent, but induced changes of feeding behaviour. The PA N-oxides showed no activity against the snails. The mixture of adenostylone and neoadenostylone was deterrent and induced feeding aversions. Deacyladenostylone was highly deterrent, but did not induce changes of feeding behaviour. At the Jura site, PA content of Adenostyles was lower than at the Black Forest site. The snails from Jura consumed much less Adenostyles than the snails from Black Forest, and also ate a little less of the treated leaf discs. The PAs which are encountered by the snails in their natural food plants (PA extract and senecionine) were more deterrent than retrorsine (a novel compound). This suggests that the snails have mechanisms for the rejection of allelochemicals which they encounter in their natural food plants, but not for novel allelochemicals. The results suggest two hypotheses regarding the function of the allelochemicals in Adenostyles: (1) The allelochemicals act mainly on very young snails. (2) PAs render Adenostyles toxic, while STs act as feeding deterrents.     

Insect feeding deterrent and growth inhibitory activities of scopoletin isolated from Artemisia annua against Spilarctia obliqua (Lepidoptera: Noctuidae)

Abstract Artemisia annua (Asteraceae) is well known for its antimalarial activities due to presence of the compound artemisinin. We isolated a methoxy coumarin from the stem part of A. annua and confirmed its identity as scopoletin through mass spectral data. The structure was established from ¹H‐nuclear magnetic resonance (NMR), ¹³C‐NMR. The compound scopoletin was evaluated for its feeding deterrence and growth inhibitory potential against a noxious lepidopteran insect, Spilartctia obliqua Walker. Scopoletin gave FD₅₀ (feeding deterrence of 50%) value of 96.7 μg/g diet when mixed into artificial diet. S. obliqua larvae (12‐day‐old) exposed to the highest concentration (250 μg/g diet) of scopoletin showed 77.1% feeding‐deterrence. In a growth inhibitory assay, scopoletin provided 116.9% growth inhibition at the highest dose of 250 μg/g diet with a GI₅₀ (growth inhibition of 50%) value of 20.9 μg/g diet. Statistical analysis showed a concentration‐dependent dose response relationship toward both feeding deterrent and growth inhibitory activities. Artemisinin is found mainly in the leaves of A. annua and not in the stems, which are typically discarded as waste. Therefore identification of scopoletin in stems of A. annua may be important as a source of this material for pest control.     

Continuous exposure to the deterrents cis‐jasmone and methyl jasmonate does not alter the behavioural responses of Frankliniella occidentalis

Behavioural responses of Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), a generalist, cell sap‐feeding insect species with piercing‐sucking mouthparts, after continuous exposure to two deterrent secondary plant compounds are investigated. We compared in choice assays on bean leaf discs, the settling, feeding, and oviposition preferences of F. occidentalis females that had no experience with the two fatty acid derivatives methyl jasmonate and cis‐jasmone before testing (naïve thrips) vs. females that had been exposed to the deterrent compounds before testing (experienced thrips). The thrips were exposed to the deterrents at low or high concentrations for varied time periods and subsequently tested on bean leaf discs treated with the respective deterrent at either a low or a high concentration. Frankliniella occidentalis females avoided settling on the deterrent‐treated bean leaf discs for an observation period of 6 h, independent of their previous experience. Our results demonstrate that feeding and oviposition deterrence of the jasmonates to the thrips were not altered by continuous exposure of the thrips to the jasmonates. Habituation was not induced, neither by exposure to the low concentration of the deterrents nor by exposure to the high concentration. These results indicate that the risk of habituation to two volatile deterrent compounds after repeated exposure is not evident in F. occidentalis. This makes the two compounds potential candidates to be integrated in pest management strategies.     

A taste receptor neurone dedicated to the perception of pyrrolizidine alkaloids in the medial galeal sensillum of two polyphagous arctiid caterpillars

It is shown that pyrrolizidine alkaloids are phagostimulants for the caterpillars of two polyphagous arctiid caterpillars, Estigmene acrea and Grammia geneura. The caterpillars will also eat dry glass‐fibre discs containing only pyrrolizidine alkaloid ? an example of pharmacophagy. The tip‐recording technique is used to demonstrate that each species has a neurone in the medial galeal styloconic taste sensillum responding to pyrrolizidine alkaloids, although the species differ in their sensitivities. This neurone responds to at least four different pyrrolizidine alkaloids and their N‐oxides, and experiments indicate that it is dedicated to perception of these compounds. The sensory response is phasic–tonic and during the tonic phase remains unchanged for at least 500 ms, resembling the type of response often seen in neurones that are sensitive to plant secondary compounds producing deterrent effects.     

Biological activity of essential oils from seven Azorean plants against Pseudaletia unipuncta (Lepidoptera: Noctuidae)

Ten essential oils from seven Azorean plant species were evaluated for their insecticidal, ovicidal, feeding‐deterrence and growth inhibition activities against Pseudaletia unipuncta. The oils of Laurus azorica (leaves), and Juniperus brevifolia (leaves) showed strong moderate insecticidal effect on fourth‐instar larvae causing 93.3% and 46.7% mortality, respectively. Juniperus brevifolia (leaves), L. azorica (leaves), Persea indica (leaves), Hedychium gardnerianum (leaves) and Pittosporum undulatum (fruits and leaves) significantly affected the hatching of P. unipuncta eggs (<8% eclosions). Five oils showed significant feeding deterrent activities (L. azorica, 92.4%, J. brevifolia, 93.6%, P. undulatum leaves, 95.5% and fruits, 83.8% and H. gardnerianum, 88.2%). All of the essential oils tested, significantly inhibited the larval growth after 5 days of feeding on the treated diet. Essential oils from L. azorica and J. brevifolia were the most potent growth inhibitors among the oils tested, producing a decrease in the initial larva weight (?14.8 and ?14.5 mg, respectively). Our results indicate that L. azorica (leaves), J. brevifolia (leaves), P. indica (leaves), H. gardnerianum (leaves), and P. undulatum (leaves and fruits) can be exploited for the development of bioactive compounds as a new source of agrochemicals. Further emphasis on isolation and identification of active constituents can be useful to develop new environment‐friendly insect control agents.     

苏云金杆菌δ-内毒素、芽孢及苏云金素A对棉铃虫毒性及拒食性的比较

苏云金杆菌库斯塔克变种HD-1的晶体蛋白与芽孢、HD-1无晶体突变株(Cry-)的芽孢以及苏云金素A对棉铃虫Helicoverpa armigera毒性和拒食性的比较研究显示,HD-1晶体蛋白对棉铃虫的杀虫毒力高, 拒食作用强; HD-1芽孢对棉铃虫具有一定的杀虫活性和生长抑制作用, 并有很强的拒食作用; HD-1无晶体突变株(Cry-)芽孢对棉铃虫无毒也无拒食作用;苏云金素A对棉铃虫的生长发育有极显著的抑制作用, 但对棉铃虫无拒食作用,由此证明晶体蛋白是苏云金杆菌杀虫活性和拒食作用的主要来源。苏云金素A与苏云金杆菌芽孢晶体混合物一起使用, 可使棉铃虫的死亡率显著提高。     

Some properties of gustation in the larva of Pieris brassicae

The effects of some chemical substances on feeding response were studied in the larvae of Pieris brassicae L. Some synergistic effects between feeding stimulants were found. The larvae appeared to be highly sensitive to deterrents. In the medial sensillum styloconicum of the maxillae a chemoreceptor is present which is stimulated by a wide range of specific feeding inhibitors among which many belong to the alkaloids. Some physiological properties of the deterrent sensitive receptor are given and its significance in food selection behaviour is discussed.

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Résumé Les effets de quelques substances chimiques sur la prise de nourriture ont été étudiés chez la chenille de Pieris brassicae. Des effets synergiques se manifestent entre certains acides aminés ou certaines vitamines et le saccharose, lorsque ces produits sont à des concentrations appropriées. Par ailleurs les chenilles se montrent très sensibles à la présence de substances inhibant la prise de nourriture. Un neurone chimiorécepteur appartenant au sensillum styloconicum médian des maxilles se révèle sensible à une grande variété de ces substances inhibitrices, dont la plupart appartiennent aux alcaloides stéroides. Ce récepteur est sensible aux composés voisins de l'ecdysone présents dans certaines plantes, et son rôle physiologique est discuté, en rapport avec le comportement alimentaire et le choix de la nourriture.

     

Structural and chemical defenses of echinoderms from the northern Gulf of Mexico

The feeding deterrent effects of echinoderm body-wall tissues and ethanolic extracts containing mid-polarity compounds were evaluated utilizing generalist fish and crabs as model predators. The body-wall tissues of the echinoderms examined ranged 10-fold from 0.9–9.4 mm in thickness, and four and a half-fold in level of mineralization (17.8–82.7% ash content). Holothuroids had the thickest body-wall tissues and contained the lowest levels of mineralization in their body-walls. Crinoids and ophiuroids had high levels of mineralization in their arms. Asteroid body-wall tissues varied the most in thickness and ash content (0.9–3.9 mm in thickness and 29.2–55.5% in ash content). Body-wall tissues of 19 species of echinoderms were tested for their feeding deterrent properties against the marine fishes Lagodon rhomboides (Linnaeus) and Cyprinodon variegatus (Lacepede), as well as the decapod crustacean Libinia emarginata (Leach). Equivalent sized pieces of fresh body-wall tissue of 16 species of echinoderms caused observable feeding deterrence responses in at least two of the three model predators. There was no significant correlation between body-wall thickness or percent ash and its palatability to any of the three model predators. Agar pellets containing ethanolic body-wall extracts of 12 of 18 echinoderm species caused observable feeding deterrence responses in the fish L. rhomboides. In similar experiments with the arrow crab Stenorhyncus seticornis (Herbst), using carrageenan fish-meal blocks as food models, no differences in consumption of control fish-meal and experimental body-wall extract blocks were detected. Our findings indicate that invertebrate and vertebrate predators may respond quite differently to echinoderm body-wall extracts.     

Insect growth regulating and antifeedant effects of neem extracts and azadirachtin on two aphid species of ornamental plants

Leaf disc choice test bioassay demonstrated that formulated neem seed extracts were highly deterrent and growth regulatory to rose aphid,Microsiphum rosae (L.) and Chrysanthemum aphid,Macrosiphoniella sanbornii (Gillete). Effective concentrations to produce 50% feeding deterrence was 0.80 and 0.84% respectively for 2nd instar nymphs irrespective of bioassay duration. The disruption of aphid feeding was related to the presence of azadirachtin concentration in the extract. The toxicity on contact from the leaf surface or via topical application due to azadirachtin was significantly different and topical treatment was at least 7 times more effective for both species. Thus growth regulatory effects of azadirachtin were influenced by the host plant and the stage of treatment. Field evaluation with formulated neem extracts revealed the effect to be more of growth regulatory nature thereby showing that azadirachtin is a physiological toxin for aphid species. Neem seed extracts reduced the population of aphid on respective host plants significantly, EC₅₀ values being 0.88 and 0.96% forM. rosae andM. sanbornii respectively.     

Feeding responses of specialist herbivores to plant extracts and pure allelochemicals: effects of prolonged exposure

We investigated the possibility of decreased feeding deterrent response following prolonged exposure in three specialist herbivores, Plutella xylostella (larvae), Pseudaletia unipuncta (larvae), and Epilachna varivestis (adults) through leaf disc choice and no‐choice tests. Neonate larvae (<24 h old) of the three species were reared on their host plants sprayed with the antifeedants until tested. Our results demonstrated that Ps. unipuncta did not show a decrease in feeding deterrent response to extracts of Melia volkensii (choice and no‐choice tests) or oregano (Origanum vulgare) (choice test) following prolonged exposure. Plutella xylostella exhibited a decrease in feeding deterrent response to M. volkensii in a choice test only, but not to oregano. Although not significant, we did find a trend toward decreasing feeding deterrent response to M. volkensii by both species in no‐choice tests. However, both species exhibited a significant decrease in feeding deterrent response to pure allelochemicals (Ps. unipuncta to thymol, and P. xylostella to thymol and toosendanin) compared with the naïve groups, following prolonged exposure in leaf disc choice tests. Epilachna varivestis showed a significant decrease in feeding deterrent responses to both oregano and thymol in leaf disc choice tests. We conclude from our present and previous studies that not only are there interspecific differences between generalist and specialist species but also among specialist species.     

Nutritional ecology of Preuss's red colobus monkey (Colobus badius preussi Rahm 1970) in Korup National Park, Cameroon

Studies on the nutritive ecology of red colobus monkeys (Colobus badius preussi) in Korup National Park, Cameroon, investigated food plants and the parts eaten as well as nutritive values of the various food items. Food plants were identified through direct observations and collected for laboratory determination of nutrient content. Food plants were analysed for crude protein (CP), crude fibre (CF), ether extract (EE), nitrogen‐free‐extractive (NFE) and total ash (A). A total of seventeen species belonging to nine different families were identified as food plants for red colobus. Food and feeding habits showed that the species were highly folivorous, tending to prefer the young leaves of food plants. Lecomtedoxa klaineana (27.0%) and Xylopia aethiopica (22.2%) of families Sapotaceae and Annonaceae, respectively, constituted about 50% of total food items consumed. Results of nutrient composition indicated individual nutrients varied significantly (P < 0.01) with food species.     

Behavioural and sensory responses to drimane antifeedants in Pieris brassicae larvae

Fifteen drimane compounds were tested for their feeding inhibiting activity in larvae of Pieris brassicae L. (Lepidoptera: Pieridae) when applied to leaf material of the host plant Brassica oleracea L. The antifeedant efficacy of the drimanes was related to their molecular structure in order to identify important functional groups. Of the drimanes tested, those with a lactone group on the B-ring were the most effective feeding inhibitors. Additionally, the sensory responses to 13 of the drimanes were measured. Neural activity was evoked in the deterrent cell in the medial sensillum styloconicum. Also, inhibition of sensory responses to feeding stimulants was found. Results of behavioural and electrophysiological tests were correlated in an attempt to elucidate the sensory code underlying feeding inhibition by drimanes in Pieris brassicae. It was concluded that the response of the deterrent cell in the medial sensillum styloconicum contributes significantly to inhibition of feeding behaviour in larvae of Pieris brassicae.     

Food discrimination and induction of preference for artificial diets in the tobacco hornworm, Manduca sexta

ABSTRACT. Feeding discrimination of Manduca larvae reared on leaves and a variety of artificial media was tested against leaves, various media, solvent fractions of leaves and media, and media components, including pure chemicals. Field-collected animals showed the same preferences as did animals cultured in the laboratory for many generations. Rearing larvae on a leaf species, an artificial diet, or homogenized leaves added to artificial diet induced a preference for that food in subsequent choice tests. Extracts of these foods using organic solvents (particularly hexane) elicited feeding choices resembling those evoked by the foods themselves: water extracts were effective as stimulants or deterrents, but responses to them differed considerably from responses to the foods. Animals raised on a defined artificial diet showed an increased preference (or decreased deterrence) for the lipid component (linolenic acid) of that diet in behavioural tests. Thus induction of preference was shown to be influenced by a specific nutrient compound. Fifteen artificial diets were tested; three were successful, including a completely defined medium. Various components of the diets were tested for feeding preferences, both as omissions from the main diet and as pure compounds. Some were stimulatory, most were neutral or slightly deterrent; a few were strongly deterrent. With the exception of linolenic acid, preferences for pure compounds were not significantly correlated with the food on which the animals were reared. The data suggest that food discrimination depends on the perception of a complex chemical message comprised of both polar and non-polar compounds, with the latter being of somewhat greater importance. The results are consistent with the explanation of the induction of feeding preferences being due to a reduced responsiveness to deterrents, to an increased preference for feeding stimulants or to both.