Electrophysiological studies of gustation in lepidopterous larvae

Summary Comparisons were made of the eleotrophysiological responses of the maxillary gustatory receptors of the following categories of caterpillars: (1) three closely related species (Papilio polyxenes L.,P. troilus L., andP. glaucus L.) each of which feeds on a different group of plants; (2) two unrelated oligophagous species (P. glaucus L. andMalacosoma americana Fabr.) that have one preferred food plant in common; (3) three unrelated monophagous species (Danaus plexippus L.,Euchaetias egle Drury, andPygarctia eglenensis Clemens) that share the same plant. Materials tested included sodium chloride, carbohydrates, amino acids, glycosides, and the saps ofDaucus carota L. andFoeniculum vulgare Mill. (the food plants ofP. polyxenes),Sassafras albidum (Nutt.) andLindera Benzoin (L.) (the food plants ofP. troilus),Prunus virginiana L. (a favored food ofP. glaucus andM. americana),Asclepias syriaca L. andApocynum androsaemifolium L. (eaten byD. plexippus,E. egle, andP. eglenensis), andBrassica oleraceae L. (food plant ofPieris rapae).The following conclusions were drawn: (1) no species of caterpillar gives a single standard electrophysiological response to all of the plants it rejects; that is, rejection is not a unitary modality; (2) a plant that is unacceptable to several species of caterpillars does not elicit the same pattern of response from each; (3) a food plant that is shared by several species of caterpillars does not elicit the same pattern of response from each; (4) a species of caterpillar that has more than one food plant does not generate the same sensory pattern to each; (5) there is no universal difference between sensory patterns for acceptance and those for rejection.A model based upon the hypothesis of across-fiber patterning is proposed to explain these results. The essence of this model is that the receptors have unique but overlapping action spectra and that each compound or mixture of compounds in leaf saps that can be discriminated generates a unique total pattern of response. Whether or not a plant is ingested depends, therefore, not on the presence or absence of a single stimulant or deterrent but upon the total sensory impression derived from integrated response to multiple plant components. Prior to the first bite a caterpillar makes its first discrimination on the basis of olfactory clues.This work was supported by National Science Foundation Grant GB 1472 and the Class of 1877 Professorship of Princeton University.     

Olfactory coding by lepidopterous larvae

Extracellular electrophysiological recording from olfactory receptors in the antennae of tobacco hornworm larvae (Manduca sexta (Johan.)) has revealed that cells respond differentially to different odors by changing latency, rate of increase of frequency of firing, rate of adaptation, and alternation of frequency increase and decrease. The resulting temporal patterns of spike activity could function as a code to allow for discrimination among various plant odors.

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Zusammenfassung Extrazelluläre elektrophysiologische Erregungsableitungen von Geruchsrezeptoren in den Antennen von Tabakschwärmer-Raupen (Manduca sexta (Johan.)) ergaben, daß die Zellen auf verschiedene Duftstoffe unterschiedlich mit Änderung der Latenzzeit, der Zunahmerate der Erregungsfrequenz, der Anpassungsrate sowie der Änderung der Frequenz-Zunahme und-Abnahme reagieren. Die sich daraus ergebenden Zeitmuster der Spike-Aktivität könnten als Code dienen und so die Unterscheidung zwischen verschiedenen Pflanzendüften ermöglichen.

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This research was supported by National Science Foundation Grant GB-1472.     

Activation of 14C-dimethoate in lepidopterous larvae

1. Dimethoate was generally more toxic when ingested than when applied topically to similar sized larvae of Mamestra brassicae (L) (4th-instar), Spodoptera littoralis (Boisd) (4th-instar), Pieris brassicae (L) (3rd-instar) and Bombyx mori (L) (2nd-instar).2. In vivo oxidation of ingested ¹⁴C-dimethoate to omethoate decreased in the order of S. littoralis > P. brassicae > B. mori and followed the order of decreasing susceptibility.3. ¹⁴C-dimethoate was regurgitated and excreted and was related to feeding rates of the species.4. The implications of the above data in relation to causes of variation in interspecies susceptibility with particular reference to concepts of resistance as influenced by polyphagy versus monophagy are discussed.     

Phospholipase activity in gut juice of lepidopterous larvae

Phospholipase A activity has been identified in gut juice from larvae of Pieris brassicae, Spodoptera exiguens, and Trichoplusia ni. The specific activity is high and suggests a rôle in lipid breakdown and assimilation.     

Induction of specific food preference in lepidopterous larvae

Last instar larvae of Manduca sexta (Johanssen) and Heliothis zea (Boddie), fed on different host plants or on artificial diet, and then tested individually, have shown clear preference for the plant previously eaten. This induced preference is specific for the inducing plant species and is not merely a change in the insect's general threshold of food acceptability. The extent to which preference can be induced by various host plants differs considerably. No induction is possible with plants outside the insect's host range. In larvae fed on artificial diet preference to a given host plant can be induced even by a one-day feeding on it. A preference once induced is not wiped out by two larval moults and subsequent feeding on an artificial diet. Thus it is supposed that the information serving as a basis for the induced feeding habit is stored in the central nervous system. The possible ecological significance of the induced preference is discussed.

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Zusammenfassung Larven des letzten Entwicklungsstadium von Manduca sexta (Sphingidae) und Heliothis zea (Noctuidae), die an verschiedenen Wirtspflanzen gezüchtet wurden, bevorzugten in individuellen Nahrungswahlversuchen jene Pflanzenarten, an welchen sie sich entwickelten. Diese induzierte Präferenz ist spezifisch für die induzierende Pflanzenart und nicht einfach eine allgemeine Verschiebung der Reizschwelle für Frass- bzw. Vergällungsstoffe. Die Stärke der durch verschiedene Wirtspflanzen induzierten Präferenz variierte von Pflanze zu Pflanze erheblich. Es war nicht möglich, Präferenz für eine Pflanzenart ausserhalb des Wirtspflanzenkreises zu induzieren. In den an künstlicher Nahrung gezüchteten Raupen entwickelte sich die Präferenz für eine gegebene Wirtspflanze bereits während einer eintägigen Ernährung an derselben. Die einmal erworbene Präferenz wurde selbst durch zwei Häutungen und Ernährung mit künstlicher Nahrung nicht ausgelöscht. Demnach wird angenommen, dass die als Basis für die induzierte Präferenz dienende Rezeptoreninformation im Zentralnervensystem gespeichert wird. Die wahrscheinliche ökologische Bedeutung der induzierten Präferenz wird besprochen.

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Supported by National Institutes of Health grant NB 07069-01.     

Gustation and foodplant selection in some lepidopterous larvae

The majority of the chemoreceptory cells, which are located in the sensilla styloconica on the maxillae, have been characterized in two oligophagous caterpillars. Besides the sugar and salt receptors M. sexta has a cell, which is stimulated by some alkaloids, a cell sensitive to some glycosides and two inositol sensitive cells. P. brassicae has, besides a sugar and a salt cell two cells sensitive to mustard oil glucosides (differing in their sensitivity patterns), one cell sensitive to amino acids and another one to anthocyanins. In M. sexta, the sensitivity of some cells to their specific stimuli can be reduced by adding these specific compounds to its food. Modifications of food selection behaviour can be correlated with such induced receptor changes. The consequences of these observations are discussed in relation with food conditioning phenomena.

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Résumé La plupart des récepteurs gustatifs, localisés dans des sensilla styloconica sur les maxilles, ont été caractérisés dans deux espèces de chenilles oligophages. En dehors des récepteurs pour le sucre et les sels, Manduca sexta a une cellule, qui est stimulée par quelques alcaloides, une autre sensible à quelques glycosides et deux cellules qui sont sensibles à l'inositol. La chenille de Pieris brassicae possède, en dehors des récepteurs pour le sucre et les sels, deux cellules qui réagissent aux glucosides des sénévols, une cellule sensible aux acides aminés et une autre aux anthocyanines. Le type de réaction induit par la sève d'une plante n'est pas seulement compliqué par des interactions inhibitrices et synergistes entre les divers composants de la séve, mais aussi par le fait que la sensibilité de certains chimiorécepteurs dépend en quelque sorte de la composition de la nourriture. Les modifications de sensibilité des récepteurs sont accompagnées par des changements du comportement dans la sélection des plantes-hôtes.

     

Effect of avian predation on survival of leaf-rolling lepidopterous larvae

Manipulative field experiments were performed in a northern Japanese deciduous forest to determine the role of avian predation in the survival of three leaf-rolling lepidopterous larvae (Homonopsis foedenratana, Archips viola, and Zeiraphera corpulentana) on Japanese lilac (Syringa reticulata) trees. When all leaf rolls on the lilacs were experimentally removed, the visitation rate of birds declined, indicating that the leaf rolls served as a visual cue for the birds. In a second experiment in which birds were excluded from the lilac trees, the survival of H. foedenratana larvae was improved, whereas the mortality of the other two Lepidoptera stayed the same. This is probably the result of differences in the shape of leaf rolls; larvae of H. foedenratana alter the leaf form to a greater extent, which may facilitate the detection of the rolls by birds. The high predation rate on the tight leaf rolls of H. foedenratana may be offset by enhanced resistance against other predators and better food quality for the enclosed larvae. Received: January 30, 1998 / Accepted: December 2, 1998     

The tolerance of lepidopterous larvae to an insect selective neurotoxin

The interaction of the fast-neurotoxic and insect selective polypeptide derived from scorpion venom (AaIT) with lepidopterous larvae tissues was studied through assays of toxicity, chromatography, binding and light microscopical autoradiography. The native and/or radioiodinated toxin was shown to:

• 1.(1) Induce a delayed, slow, progressive paralysis (within 24–48 h) of Spodoptera larvae by relatively high doses (paralytic unit = 2.4 μg/100 mg) corresponding to about only 10% of the total toxicity of the crude venom. Larvae of six species representing five families of Lepidoptera responded similarly to the toxin.
• 2.(2) Resist an in vitro incubation in the insect's hemolymph.
• 3.(3) Lose 80% of its toxicity in the insect's body within 24 h, accompanied by a progressive process of degradation and elimination by the excretory system.
• 4.(4) Specifically bind to a single class of non-interacting binding sites of high affinity and low capacity (0.2 pmol/mg protein, similar to tritiated saxitoxin) in an in vitro, homogenate derived, neuronal preparation.
• 5.(5) Specifically bind with high affinity to desheathed but otherwise intact nerves.
• 6.(6) Be devoid of accessibility to peripheral-terminal branches of Spodoptera motor nerves in situ—strongly contrasting those of the toxin susceptible Periplaneta nerves.

It may be thus concluded that the tolerance of the lepidopterous larvae to AaIT can be substantially attributed to pharmacokinetic aspects of toxin accessibility barriers and degradation processes.     

Regulation and significance of ecdysteroid titre fluctuations in lepidopterous larvae and pupae

The last larval moult of Galleria mellonella is induced by an elevation of ecdysteroid titre to more than 200 ng/g. After ecdysis the titre remains very low until 70 hr of the last-instar when a slight elevation in ecdysteroid concentration initiates the onset of metamorphosis. An ecdysteroid peak (275 ng/g), which occurs between 108 and 144 hr, is associated with wandering and cocoon spinning. Pupal ecdysis follows about 20 hr after a large ecdysteroid peak (780 ng/g) with a maximum in slowly-mobile prepupae (160 hr of the last larval instar). The ecdysteroid decrease between the two peaks coincides with the period when the larvae exposed to unfavourable conditions enter diapause. The pupal-adult moult is initiated by a high ecdysteroid peak (1500–2500 ng/g) in early pupae and imaginal cuticle is secreted in response to a smaller peak (ca. 500 ng/g) in the middle of pupal instar.Until early pupae, the ecdysteroid content is regulated by the prothoracic glands. In decapitated larvae the glands become spontaneously active after 30–40 days and the body titre of ecdysteroids undergoes an increase; the glands revert to inactivity when the insects accomplish secretion of pupal cuticle. A similar ecdysteroid increase occurs within 10 days when the decapitated larvae receive implants of brains releasing the prothoracicotropic neurohormone (PTTH). In either case, the pupation-inducing increase of ecdysteroids is 3 times higher than the large ecdysteroid peak in the last-instar of intact larvae. This indicates that the function of prothoracic glands in intact larvae is restrained, probably by the juvenile hormone (JH). Exogenous JH suppresses the spontaneous activation of the prothoracic glands in decapitated larvae and reduces the ecdysteroid concentration in those larvae (both decapitated and intact), whose glands were activated by PTTH. Furthermore, JH influences the PTTH release from the brain in situ: depending on JH concentration and the age and size of treated larvae, the PTTH liberation is either accelerated or delayed.Neither in G. mellonella larvae, nor in the diapausing pupae of Hyalophora cecropia and Celerio euphorbiae, does JH directly activate the prothoracic glands. It is suggested that the induction of the moult by JH in decerebrate insects, which has been observed in some species, is either due to indirect stimulation of ecdysteroid production or to increased sensitivity of target tissues to ecdysteroids. In G. mellonella, a moult occurs at a 5–15 times lower than usual ecdysteroid concentration when the last-instar larvae are exposed to JH.     

Organization of the retrocerebral gland system in lepidopterous larvae of the family pyralidae

The organization of the retrocerebral gland system in larvae of six species of Lepidoptera belonging to the family Pyralidae was compared using light and electron microscopy. We have demonstrated for the first time the presence of separate corpora cardiaca and corpora allata in the following economically important borers: the southwestern corn borer, Diatraea grandiosella, the sugar cane borer, Diatraea saccharalis, the European corn borer, Ostrinia nubilalis, and the rice stalk borer, Chilo plejadellus. In these species a long nervus corporis allati (ca. 300 μm) runs from the corpus cardiacum to the corpus allatum which is attached to the duct of the mandibular gland.The identity of the corpora allata of D. grandiosella was confirmed by transplantation. Corpora allata removed from pre-diapausing larvae and implanted into the haemocoele of early last stage non-diapausing larvae led to a high incidence of supernumerary larval rather than pupal ecdyses.     

Antifeedant effects of azadirachtin and structurally related compounds on lepidopterous larvae

The antifeedant activity of azadirachtin, azadirachtin-derivatives and related limonoids was assessed in choice and no-choice bioassays against four species of Lepidoptera: Spodoptera littoralis, Spodoptera frugiperda, Heliothis virescens and Heliothis armigera. The choice bioassay showed that the feeding behaviour of S. littoralis was affected by more of the compounds than that of either S. frugiperda or H. virescens. H. armigera was the least affected. Azadirachtin and dihydroazadirachtin were the most potent of the 40 compounds tested. The results showed that hydrogenation of the C-22,23 double bond did not decrease antifeedant activity and the nature of the substitutes at C-1, C-3 and C-11 were important. Molecules with bulky substitutes at either C-22 or C-23 were usually ineffective antifeedants as were compounds lacking an epoxide. Compounds recorded as active antifeedants in the choice bioassay were not always as active in the no-choice test. The value of the bioassays in assessing the mode of action of the compounds is discussed.

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Résumé L'activité phagodissuadante de l'azadirachtine, de ses dérivés et des limonoïdes voisins sur 4 espèces de lépidoptères: Spodoptera littoralis, S. frugiperda, Heliothis virescens et H. armigera a été évaluée par des expériences avec et sans choix. Les expériences de choix ont montré que le comportement alimentaire de S. littoralis était modifié par plus de substances que celui de S. frugiperda ou H. virescens. Celui de H. armigera était le moins modifié. Les 2 substances les plus puissantes parmi les 40 examinées, ont été l'azadirachtine et le dihydroazadirachtine. Ces résultats montrent que l'hydrogénation de la double liaison C-22,23 ne réduit pas l'activité phagodissuadante et que la nature des substitutions en C-1, C-3 et C-11 sont importantes. Les molécules avec des substitutions volumineuses en C-22 ou C-23 sont généralement des phagodissuadants aussi inefficaces que ceux ayant perdu un époxide. Les substances notées comme phagodissuadants actifs dans les expériences de choix ne sont pas toujours aussi actives en absence de choix. La valeur des tests dans l'évaluation du mode d'action des substances est discuté.

     

The sensory basis of trail following in some lepidopterous larvae: contact chemoreception

ABSTRACT. . Caterpillars of the genus Malacosoma follow trails of the chemical 5-beta-cholestane-3,24-dione, but nothing is known of how they perceive this compound, or more generally about the sensory basis of trail following in caterpillars. By selective ablations of chemosensory organs we show that, in Malacosoma , the trail chemical is perceived by the maxillary palpi. In another lepidopteran species, Yponomeuta cagnagellus , the palpi are needed to discriminate their own trails from a trail of Malacosoma. Malacosoma larvae also lose their specificity for conspecific trails when their palpi are ablated. Volatile cues evidently do not play a role in trail-following behaviour, since neither Malacosoma nor Yponomeuta can orient on a trail covered with fine nylon mesh. These data indicate that for Malacosoma , and probably also for Yponomeuta , contact chemoreception mediated by the maxillary palpi is the primary mode of pheromone perception. The evolution of receptor sensitivity to trail chemicals in caterpillars is discussed.     

Azadirachtin: structural requirements for reducing growth and increasing mortality in lepidopterous larvae

Spodoptera littoralis, Spodoptera frugiperda and Heliothis virescens were exposed to azadirachtin and 23 other related compounds in three bioassays: oral cannulation, haemolymph injection and topical application. Spodoptera littoralis was susceptible to a larger proportion of the compounds than either of the other species. Overall, azadirachtin, 22,23-dihydroazadirachtin and 1-detigloyl-22,23-dihydroazadirachtin were the most active compounds. The nature of the substitutes at C-1 and C-3 of the decalin ring affects the potency of the compounds, as does the addition of substitutes to C-22,23. Generally, larvae were less sensitive to compounds when they were topically applied than when they were cannulated into the gut or injected into the haemolymph.

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Résumé Spodoptera littoralis, S. frugiperda et Heliothis virescens ont été exposés à l'azadirachtine et 23 autres substances voisines dans 3 expériences: canulation orale, injection d'haemolymphe et application topique. S. littoralis est sensible à un plus grand nombre de substances que les 2 autres espèces. De toutes façons, les substances les plus actives ont été l'azadirachtine, la 22,23-dihydroazadirachtine et la 1-détigloyl-22,23-dihydroazadirachtine. La nature des substitutions en C-1 et C-3 de l'anneau décaline modifie la puissance des substances, tout comme les substitutions en C-22,23. D'une façon générale, les chenilles ont été moins sensibles aux applications topiques qu'aux canulations dans le tube digestif ou aux injections dans l'haemolymphe.

     

Use of feeding rate as an indicator of caloric value in some lepidopterous larvae

Summary A significant, positive correlation between feeding rate and caloric value becomes apparent in 6 species of lepidopterous larvae exposed to different rations, food plants and temperatures. There is a possibility of using feeding rate of the final instar larvae as an indicator of its caloric value. Larvae feeding at the rates of 100, 200, 400 and 600 mg dry weight/g live insect/day contained around 5,320, 5,670, 5,800 and 5,900 g cal/g dry weight, respectively.     

Use of larval weight and length as indices of gut content weight in some lepidopterous larvae

Summary Significant correlation was obtained between body weight or length and the weight of contents of gut in chosen lepidopterous larvae ranging from 50 to 3,300 mg or 13 to 75 mm. The obtained relations can be used in the determination of net weight of a larva (i.e. the weight of the larva excluding its gut contents) and hence may find wide application in ecological studies.     

Diversity of culturable phyllosphere bacteria on beech and oak: the effects of lepidopterous larvae

The community composition of epiphytic heterotrophic bacteria on leaves of beech and oak, which were either damaged by lepidopterous larvae or remained undamaged, was investigated. In addition, the ability of these bacteria to utilize inorganic nitrogen was studied. The bacteria were isolated on nutrient agar and systematically identified with biochemical and physiological tests. Rarefaction plots and the Shannon-Wiener function revealed that species diversity was significantly higher on leaves of damaged beech compared to undamaged leaves, but no differences were found on leaves of oak. The portion of bacterial isolates showing a strong response to ammonia and nitrate was significantly larger on leaves of oak than on those of beech. Furthermore, significantly more isolates with a high capability to assimilate both nitrogen compounds were found on leaves attacked by the folivorous larvae compared to those not attacked on oak. It is suggested that the changes in the microbial community in response to folivorous insects might affect the extent of nutrient cycling exceeding eventually the scale of a leaf.