The endocrine basis for developmentally stationary prepupae in larvae ofTrichoplusia ni pseudoparasitized byChelonus insularis

Summary Larvae of the cabbage looper,Trichoplusia ni, precociously initiate metamorphosis in the penultimate instar when parasitized byChelonus insularis. Some larvae developing from stung eggs precociously spin cocoons, but upon dissection contain no live or obvious parasites. Such pseudoparasitized larvae greatly slow down in development as prepupae, due to a suppressed ecdysteroid titer which in turn may be caused by a suppressed juvenile hormone titer.Abbreviations JH juvenile hormone - JHA juvenile hormone analog - JHE juvenile hormone esterase     

Precocious expression of the final larval instar developmental pattern in larvae of Trichoplusia ni pseudoparasitized by Chelonus spp

The present study has used a number of electrophoretic approaches to analyze the proteins in normal Trichoplusia ni larvae and those pseudoparasitized by Chelonus spp. A number of feeding-stage, hemolymph proteins appear or increase dramatically only during the final larval stadium. Other proteins highly abundant only during the penultimate stadium disappear or decrease dramatically during the final stadium. The comparative protein profiles of penultimate instar, pseudoparasitized larvae are very similar to those of last instar larvae. These changes in hemolymph proteins are seen on gels resulting from electrophoresis, isoelectric focusing (wide range and narrow range Ampholine and very narrow range Immobiline gels) and SDS-disc electrophoresis. It is concluded that the entire last instar developmental pattern of protein gene products is occurring precociously in pseudoparasitized larvae.     

Parasite interaction with host complement: beyond attack regulation

Many orthologous proteins of known mammalian receptors have been discovered in parasites. Besides disguising the parasite as self in terms of the host immune system, evidence is accumulating that these receptors link to signalling pathways in parasites that appear to be involved in their growth or development. Recently, several proteins of the host complement system, which forms part of the innate defence against invading microorganisms, have been shown to possess alternative functions. These complement proteins interact with signalling pathways involved in early development and differentiation, as well as organ and tissue regeneration. By altering cellular interactions and responses, complement is being shown to have novel roles besides the originally described inflammatory role. The possibility exists that, as for other host factors interacting with parasites and affecting their growth or development, host complement proteins could also have such an influence.     

Hemolymph concentrations of host ecdysteroids are strongly suppressed in precocious prepupae of Trichoplusia ni parasitized and pseudoparasitized by Chelonus near curvimaculatus.

Regulation of ecdysteroid production in lepidopteran prepupae was studied using a parasitic wasp (C. near curvimaculatus) which specifically suppresses host prepupal ecdysteroid production after the induction of precocious host metamorphosis. At the developmental stage at which the hemolymph of the unparasitized metamorphosing host has its maximum titer of prepupal ecdysteroids, the hemolymph of 4th instar "truly parasitized" hosts (hosts with a surviving endoparasite) had a strongly reduced ecdysteroid titer. However, during the photophase about 12 h later, just prior to emergence of the parasite larva, an ecdysteroid peak was observed in the host hemolymph. Fourth instar pseudoparasitized prepupal hosts (in which the endoparasite was not present or died early in development) exhibited a sustained suppression in the hemolymph ecdysteroid titer. Small 5th instar pseudoparasitized hosts, which normally would molt to a 6th instar prior to metamorphosis, but which precociously attained the prepupal stage, also had a strongly reduced ecdysteroid titer. The late increase observed in truly parasitized hosts could be completely prevented by surgical removal of the parasite 24 h earlier, resulting in a titer similar to that in pseudoparasitized hosts. HPLC analysis of ecdysteroids in normal, truly parasitized, and 4th or 5th instar pseudoparasitized prepupae showed that both ecdysone and 20-OH ecdysone* were suppressed in truly and pseudoparasitized prepupae, with ecdysteroid levels being lowest in pseudoparasitized hosts. These data, and those of Brown and Reed-Larsen (Biol Contr 1, 136 [1992]), showing endoparasite secretion of ecdysteroids just prior to its emergence from the host, strongly indicate that: (1) the prepupal peak in truly parasitized hosts originates from the endoparasite, and (2) the low level of ecdysteroids in pseudoparasitized hosts results from the host's intrinsic inability to express a normal level of prepupal ecdysteroid titer. While precocious 4th or 5th instar prepupae of similar size had similarly suppressed ecdysteroid titers, smaller 4th instar prepupae had a lower ecdysteroid titer than larger, precocious 5th instar prepupae. Rare 5th instar pseudoparasitized prepupae that were of nearly normal size showed a prepupal ecdysteroid titer distinctly greater than those of the usual smaller, precocious 5th instar prepupae. The data suggest that the competence of the host to express a normal hemolymph titer of prepupal ecdysteroids is more closely correlated with the size of the prepupae than with the instar attained.     

Cladoendesis and a new look at the evolution of insect metamorphosis

Cladoendesis is a method of phylogeny analysis opposed to various matrix methods. In contrast to matrix methods, in cladoendesis phylogenetic trees are not built each time as new ones, but are reconstructed based on the previous results. Each character of each taxon is compared with its ancestral condition in the ground-plan of the higher taxon. The revealed part of the phylogeny is represented in a form of hierarchical classification. In addition to the principles of natural classification elaborated by C. Linnaeus, P.-A. Latreille, and others, evolutionary theory elaborated by Ch. Darwin and others, and principles of phylogenetic systematics elaborated by W. Hennig and others, cladoendesis includes rank-free dual nomenclature system (DualNom) and advanced method of text layout elaborated by the author. Important components of cladoendesis are the use of the term “plesiomorphon” and taking into account such important evolutionary events as conservation and deconservation of characters. Cladoendesis allowed the author to make comprehensive revision of the phylogeny of Ephemeroptera, find homology in the structure of the maxillae of adult Aphaniptera, larval maxillae of Neuropteroidea-Birostrata, larval legs of Nannomecoptera, and genital parts of some insect taxa. It also allows understanding the nature and evolution of insect metamorphosis. It is generally believed that initially insect ontogenesis proceeds the shortest way, while such phenomena as substitutions of organs by means of their atrophy and subsequent restoring are secondary. In contrast to this, the flagellum of the antenna of Amyocerata initially grows by addition of proximal segments and simultaneous loss of distal ones; in many taxa, including Metabola (insects with complete metamorphosis), distal segments are not aborted. Some authors tried to explain the origin of complete metamorphosis by various reasons: ecological, morphogenetic, or others. This approach is wrong because all insects with complete metamorphosis constitute the holophyletic taxon Metabola Burmeister 1832, i.e., originate from a single ancestral species which acquired this kind of metamorphosis. If complete metamorphosis could appear in response to some factor, it would appear many times in different species, in which case the taxon Metabola, characterized by complete metamorphosis, would be polyphyletic. The holophyly of Metabola is well proven by cladoendesis but cannot be revealed by any matrix method. Based on understanding of these facts, the author was able to discover the specific features which appeared in the common ancestor of Metabola and then became conserved and were inherited by all its descendants. These features include loss of the scape in the larval antenna (leading to a peculiar transformation of antennae in metamorphosis) and a peculiar mode of leg transformation during molt from larva to pupa. During the larval/pupal molt, the leg loses musculature and gets an immobile knee bend, so that the pupa is unable to use its legs. This conserved feature determines the inactive mode of life of the pupae of most insects. It is usually believed that male coccids (Gallinsecta De Geer 1776) have metamorphosis similar to the complete metamorphosis of Metabola. But the phylogenetic position of Gallinsecta and Metabola, as revealed by cladoendesis, does not allow one to assume common modifications in their metamorphoses. Now, when concrete autapomorphies of Metabola have been found, it becomes possible to compare metamorphoses of Metabola and Gallinsecta. Examination of Orthezia urticae and some other coccids shows that metamorphosis of their males shares no modifications with the true complete metamorphosis. The inactive mode of life of their nymphs is not connected with any anatomical reason, but purely with the fact that in the course of transformation from the feeding wingless larva to the non-feeding winged adult, the nymph has already lost its mouth apparatus and has not yet got functional wings, so it does not need to move. At the same time, actively feeding stages of Gallinsecta, both males and females, have an unusual mode of molt transformation of the legs and antennae; this is a unique autapomorphy of Gallinsecta not found in any other insect taxon.     

A new form of insect cuticle

A hitherto unnoticed, harder form of cuticle, which occurs on the mandibles of the Australian plague locust, Chortoicetes terminifera , is described     

Human interleukin-2 production in insect (Trichoplusia ni) larvae: effects and partial control of proteolysis

Many eukaryotic proteins have been successfully expressed in insect cells infected with a baculovirus in which the foreign gene has been placed under the control of a viral promoter. This system can be costly at large scale due to the quality of virus stock, problems of oxygen transfer, and severity of large-scale contamination. To circumvent this problem, we have investigated the expression of a foreign protein, human interleukin-2 (IL-2), in insect larvae, Trichoplusia ni, infected with the baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV). The IL-2 gene was placed under control of the p10 promoter so that the polyhedra remained intact for efficient primary infection. From our results, it was clear that early infection limited larval growth and late infection delayed product production until near pupation, hence infection timing was important. Also, the harvest time was crucial for obtaining high yield, because IL-2 production had a sharp optimal peak with a time of occurrence dependent on both temperature and the initial amount of infection virus. Specifically, we found that, by raising the infection temperature to 30 degrees C, we more than doubled the protein productivity. Furthermore, a significant concern of the larvae/baculovirus expression system has been the large amount of protease produced by the larvae, which adversely affects the protein yield. Therefore, we screened several protease inhibitors and characterized the larval protease specificity and timing to attenuate their impact. This report elucidates and delineates the factors that most directly impact protein yield in the larvae expression system, using IL-2 as a model.     

昆虫的变态发育研究

昆虫变态发育使得昆虫成为地球陆地上种类最多、数量最大、分布最广、生活环境最多样化的一群生物。变态使昆虫在其生命周期中的不同发育时期表现出完全不同的形态、结构、功能和生活习性的变化,有利于昆虫迁飞转移,扩大其求偶交配、生活和生存环境空间。昆虫变态发育的变化是长期自然环境适应、协同进化的结果,受激素、营养和基因的精确调控。本文简要介绍了昆虫变态的类型、激素调控、营养调控和基因调控方面的研究进展,以及研究昆虫变态发育的科学和应用意义。     

Phenotypic plasticity and nutrition in a phytophagous insect: consequences of colonizing a new host

The European rose-hip fruit fly Rhagoletis alternata (Diptera, Tephritidae) infests hips of Rosa species. This fly includes R. rugosa, an Asian species now cultivated all over Europe, in its host range. Differences in size and biomass of hips between the ancestral host R. canina and the new host translate into better growth, shorter larval development of larvae within hips of R. rugosa and larger body size and fertility of flies which developing in the new host. In turn this causes different interactions with other organisms of the food-web centred on the host plant. The importance of nutrition and phenotypic plasticity is twofold: they generate a considerable part of life-history diversity within a species and reinforce differences in the ecological context of the ancestral and new host.     

Parasite strain coexistence in a heterogeneous host population

Heterogeneity in host susceptibility and transmissibility to parasite attack allows a lower transmission rate to sustain an epidemic than is required in homogeneous host populations. However, this heterogeneity can leave some hosts with little susceptibility to disease, and at high transmission rates, epidemic size can be smaller than for diseases where the host population is homogeneous. In a heterogeneous host population, we model natural selection in a parasite population where host heterogeneity is exploited by different strains to varying degrees. This partitioning of the host population allows coexistence of competing parasite strains, with the heterogeneity-exploiting strains infecting the more susceptible hosts, in the absence of physiological tradeoffs and spatial heterogeneity, and even for markedly different transmission rates. In our model, intermediate-strategy parasites were selected against: should coexistence occur, an equilibrium is reached where strains occupied only the extreme ends of trait space, under appropriate conditions selecting for lower R₀.     

Developmental neuroethology of insect metamorphosis

During metamorphosis, the insect nervous system must change to accomodate alterations in body form and behavior. Studies primarily on moths have shown that these changes involve the death of some larval neurons, the conservation and remodeling of others, and the maturation of new, adult-specific cells. The motor and sensory sides of the adult CNS vary in this regard with the former being constructed primarily from remodeled larval components, whereas the latter arises primarily from new neurons. Neuronal remodeling has received considerable attention. Larval-specific dendritic fields are pruned back during the larval–pupal transition, followed by the sprouting of adult-specific dendrites. Simple reflexes have been used to correlate these neuronal changes with the acquisition or loss of particular behaviors. The loss of the proleg retraction reflex is associated with the regression of the dendritic arbors of the proleg motoneurons. By contrast, expansion of axon arbors of the gin-trap afferents is necessary, but not sufficient, for the assembly of the gin-trap reflex in the pupal stage. The stretch receptor reflex provides a third example in which a new dendritic field in the adult form of a neuron is associated with new adult-specific connections. Interestingly, these connections are masked by persisting larval contacts until the emergence of the adult moth. For the metamorphosis of more complex behavioral circuits, some, such as that for flight behavior, seem to be assembled de novo, whereas others, like that for adult ecdysis behavior, show conservation of some circuit elements from the larval stage but with the superposition of some adult-specific components. © 1992 John Wiley & Sons, Inc.     

Chelonus sp.: suppression of host ecdysteroids and developmentally stationary pseudoparasitized prepupae

When eggs of the caterpillar Trichoplusia ni are stung by Chelonus sp. (near C. curvimaculatus) (Braconidae), the developing host larvae precociously spin a cocoon but then remain developmentally stationary in the prepupal stage. The latter event happens even in hosts which were stung and precociously spin cocoons but which, upon dissection, contain no obvious parasite. Injection of radiolabeled ecdysone into either pseudoparasitized or allatectomized larvae demonstrates suppressed rates of conversion of ecdysone to 20-hydroxyecdysone when compared with controls. The data indicate that the occurrence of developmentally stationary pseudoparasitized prepupae is due to less production of ecdysteroid and less conversion of ecdysone to 20-hydroxyecdysone, both probably as a result of suppressed juvenile hormone titer.     

Induction and regulation of metamorphosis in planktonic larvae:Phoronis mülleri (Tentaculata) as archetype

The larvae ofPhoronis mülleri are comprised of many diverse behavioural forms that can be manipulated experimentally to facilitate precise assertions about the induction of metamorphosis. Various parameters for inducing metamorphosis as exemplified inPhoronis, such as species-specific substrate, bacteria, the cations Rb⁺, Cs⁺ and Hg²⁺ and tensides, are considered, and their ecologic relevance to natural factors in the sea is demonstrated. Findings on metamorphosis in other marine larvae are summarized. The function of marine bacteria as “ecological ushers” is particularly emphasized.     

Developmental neuroethology of insect metamorphosis.

During metamorphosis, the insect nervous system must change to accomodate alterations in body form and behavior. Studies primarily on moths have shown that these changes involve the death of some larval neurons, the conservation and remodeling of others, and the maturation of new, adult-specific cells. The motor and sensory sides of the adult CNS vary in this regard with the former being constructed primarily from remodeled larval components, whereas the latter arises primarily from new neurons. Neuronal remodeling has received considerable attention. Larval-specific dendritic fields are pruned back during the larval-pupal transition, followed by the sprouting of adult-specific dendrites. Simple reflexes have been used to correlate these neuronal changes with the acquisition or loss of particular behaviors. The loss of the proleg retraction reflex is associated with the regression of the dendritic arbors of the proleg motoneurons. By contrast, expansion of axon arbors of the gin-trap afferents is necessary, but not sufficient, for the assembly of the gin-trap reflex in the pupal stage. The stretch receptor reflex provides a third example in which a new dendritic field in the adult form of a neuron is associated with new adult-specific connections. Interestingly, these connections are masked by persisting larval contacts until the emergence of the adult moth. For the metamorphosis of more complex behavioral circuits, some, such as that for flight behavior, seem to be assembled de novo, whereas others, like that for adult ecdysis behavior, show conservation of some circuit elements from the larval stage but with the superposition of some adult-specific components.     

Recombinant protein production in insect larvae: host choice,tissue distribution,and heterologous gene instability

The expression of the fluorescent protein, DsRed, facilitates the optimization of protein production in orally-infected whole larvae. Trichoplusia ni was shown to be a much better host for recombinant AcMNPV compared to four other noctuid Lepidopteran species achieving 100% infectivity at the minimal tested dose. The highest density of marker protein was found in endothelial and tracheal cells, fat body, and hemocytes. Trichoplusia ni larvae possessed visually detected color over sequential passages of oral infection until the sixth round. Western blot analysis confirmed the progressive decrease of both tetramer and monomer forms of DsRed. The intact DsRed gene and promoter region was present in late passages, but viral population carrying the heterologous gene had dropped more than 2-logs after the fifth round while the amount of total viral DNA remained unchanged over sequential passages. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Parasite invasion following host reintroduction: a case study of Yellowstone's wolves

Wildlife reintroductions select or treat individuals for good health with the expectation that these individuals will fare better than infected animals. However, these individuals, new to their environment, may also be particularly susceptible to circulating infections and this may result in high morbidity and mortality, potentially jeopardizing the goals of recovery. Here, using the reintroduction of the grey wolf (Canis lupus) into Yellowstone National Park as a case study, we address the question of how parasites invade a reintroduced population and consider the impact of these invasions on population performance. We find that several viral parasites rapidly invaded the population inside the park, likely via spillover from resident canid species, and we contrast these with the slower invasion of sarcoptic mange, caused by the mite Sarcoptes scabiei. The spatio-temporal patterns of mange invasion were largely consistent with patterns of host connectivity and density, and we demonstrate that the area of highest resource quality, supporting the greatest density of wolves, is also the region that appears most susceptible to repeated disease invasion and parasite-induced declines. The success of wolf reintroduction appears not to have been jeopardized by infectious disease, but now shows signs of regulation or limitation modulated by parasites.