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The origins of insect metamorphosis.

Insect metamorphosis is a fascinating and highly successful biological adaptation, but there is much uncertainty as to how it evolved. Ancestral insect species did not undergo metamorphosis and there are still some existing species that lack metamorphosis or undergo only partial metamorphosis. Based on endocrine studies and morphological comparisons of the development of insect species with and without metamorphosis, a novel hypothesis for the evolution of metamorphosis is proposed. Changes in the endocrinology of development are central to this hypothesis. The three stages of the ancestral insect species-pronymph, nymph and adult-are proposed to be equivalent to the larva, pupa and adult stages of insects with complete metamorphosis. This proposal has general implications for insect developmental biology.

Animals↗

Structural genomics sheds light on protein functions and remote homologs across the insect tree of life.

Protein structure bridges the sequence-function relationship, enabling deep exploration of biological processes across diverse organisms. Insects, the most diverse animal lineage, accounting for over 50% of all described animal species, provide an exceptional system for exploring sequence-structure-function relationships. Here, we reconstructed a comprehensive and well-resolved phylogeny of 4854 insects, spanning all orders. Leveraging this framework, we created an atlas of 13.29 million predicted protein structures from 824 representative species, including 11.63 million newly predicted structures. Structural clustering revealed that proteins with divergent sequences but similar structures could be effectively grouped together. Structural similarity searches against proteins with well-characterized functions yielded annotations for 7.61 million insect proteins, including up to 14% of previously unannotated proteins. We further identified 750 million remote homologs between insect proteins, many of which trace back to ancient branches of the insect phylogeny. Remarkably, despite extensive sequence divergence, cGAS-like receptors (cGLRs) were structurally conserved across all 824 insects. Experimental assays demonstrated that these structurally identified cGLRs play a crucial role in antiviral defense in the yellow fever mosquito. Our findings highlight the significance of structural genomics for understanding protein function and evolution across the tree of life.

Animals↗

A novel lipoprotein from the hemolymph of the cochineal insect, Dactylopius confusus.

A new type of insect lipoprotein was isolated from the hemolymph of the female cochineal insect Dactylopius confusus. The lipoprotein from the cochineal insect hemolymph was found to have a relative molecular mass of 450 000. It contains 48% lipid, mostly diacylglycerol, phospholipids and hydrocarbons. The protein moiety of the lipoprotein consists of two apoproteins of approximately 25 and 22 kDa, both of which are glycosylated. Both apolipoproteins are also found free in the hemolymph, unassociated with any lipid. Purified cochineal apolipoproteins can combine with Manduca sexta lipophorin, if injected together with adipokinetic hormone into M. sexta. This could indicate that the cochineal lipoprotein can function as a lipid shuttle similar to lipophorins of other insects, and that the cochineal insect apolipoproteins have an overall structure similar to insect apolipophorin-III.

Amino Acid Sequence↗

The development and evolution of insect mouthparts as revealed by the expression patterns of gnathocephalic genes.

To understand better both the development and evolution of insect mouthparts, we have compared the expression pattern of several developmentally important genes in insects with either mandibulate or stylate-haustellate mouthparts. Specifically, we examined the expression of the proboscipedia (pb) and Distal-less (Dll) gene products as well as three regulators of pb, Sex combs reduced (Scr), Deformed (Dfd), and cap 'n' collar (cnc). These genes are known to control the identity of cells in the gnathal segments of Drosophila melanogaster and would appear to have similar conserved functions in other insects. Together we have made an atlas of gene expression in the heads of three insects: Thermobia domestica and Acheta domestica, which likely exemplify the mandibulate mouthparts present in the common insect ancestor, and Oncopeltus fasciatus, which has piercing-sucking mouth parts that are typical of the Hemiptera. At the earliest stages of embryogenesis, only the expression of pb was found to differ dramatically between Oncopeltus and the other insects examined, although significant differences were observed later in development. This difference in pb expression reflects an apparent divergence in the specification of gnathal identity between mandibulate and stylate-haustellate mouthparts, which may result from a "phylogenetic homeosis" that occurred during the evolution of the Hemiptera.

Amino Acid Sequence↗

A respiratory hemocyanin from an insect.

Insects possess an elaborate tracheal system that enables transport of gaseous oxygen from the atmosphere directly to the inner organs. Therefore, the presence of specialized oxygen-transport proteins in the circulatory system of insects has been considered generally unnecessary. Here, we show for the first time, to our knowledge, the presence of an ancestral and functional hemocyanin (Hc) in an insect. In the hemolymph of nymphs and adults of the stonefly Perla marginata, a hexameric Hc was identified, which consists of two distinct subunit types of 659 and 655 amino acids. P. marginata Hc displays cooperative oxygen binding with a moderately high oxygen affinity [(half-saturation pressure, P(50) approximately 8 torr (1 torr = 133 Pa)]. No evidence was found for the presence of Hcs in the more evolutionarily advanced holometabolan insects, suggesting that this type of respiratory protein was lost later in insect evolution. However, our results demonstrate that, in contrast to the accepted paradigm, certain basal insects have retained an ancestral blood-based mechanism of gas exchange.

Amino Acid Sequence↗

Differential expression patterns of the hox gene are associated with differential growth of insect hind legs.

Diversification of leg appendages is one of the hallmarks of morphological evolution in insects. In particular, insect hind (T3) legs exhibit a whole spectrum of morphological diversification, ranging from uniform to extremely modified. To elucidate the developmental basis of T3 leg evolution, we have examined the expression patterns of two homeotic genes, Ultrabithorax and abdominal-A (collectively referred to as UbdA), in a broad range of species. First, our results show that UbdA expression in hemimetabolous insects is localized only in specific T3 leg segments undergoing differential growth (compared to their foreleg counterparts). In contrast, in basal hexapod and insect lineages, the absence of the UbdA signal coincides with uniform leg morphology. The same situation exists in first instar larvae of holometabolous insects, in which absence of UbdA expression in the embryonic T3 legs is associated with the lack of larval T3 leg diversification. Second, there is a clear difference in the timing of expression between species with greatly enlarged T3 leg, such as crickets and grasshoppers, and species that exhibit more moderate enlargement of hind legs, such as mantids and cockroaches. In the former, the UbdA expression starts much earlier, coinciding with the elongation of T3 limb buds. In the latter, however, the UbdA expression starts at much later stages of development, coinciding with the establishment of distinct leg segments. These results suggest that diversification of insect hind legs was influenced by changes in both the spatial and temporal regulation of the UbdA expression.

Animals↗

The pupal specifier broad directs progressive morphogenesis in a direct-developing insect.

A key regulatory gene in metamorphosing (holometabolous) insect life histories is the transcription factor broad (br), which specifies pupal development. To determine the role of br in a direct-developing (hemimetabolous) insect that lacks a pupal stage, we cloned br from the milkweed bug, Oncopeltus fasciatus (Of'br). We find that, unlike metamorphosing insects, in which br expression is restricted to the larval-pupal transition, Of'br mRNA is expressed during embryonic development and is maintained at each nymphal molt but then disappears at the molt to the adult. Induction of a supernumerary nymphal stage with a juvenile hormone (JH) mimic prevented the disappearance of br mRNA. In contrast, induction of a precocious adult molt by application of precocene II to third-stage nymphs caused a loss of br mRNA at the precocious adult molt. Thus, JH is necessary to maintain br expression during the nymphal stages. Injection of Of'br dsRNA into either early third- or fourth-stage nymphs caused a repetition of stage-specific pigmentation patterns and prevented the normal anisometric growth of the wing pads without affecting isometric growth or molting. Therefore, br is necessary for the mutable (heteromorphic) changes that occur during hemimetabolous development. Our results suggest that metamorphosis in insects arose as expression of br, which conveys competence for change, became restricted to one postembryonic instar. After this shift in br expression, the progressive changes that occur within the nymphal series in basal insects became compressed to the one short period of morphogenesis seen in the larva-to-pupa transition of holometabolous insects.

Animals↗

Granary trial of protein-enriched pea flour for the control of three stored-product insects in barley.

A granary trial was conducted to evaluate the efficacy of protein-enriched pea flour against three common stored-grain insects, Sitophilus oryzae (L.), Tribolium castaneum (Herbst), and Cryptolestes ferrugineus (Stephens). Six 30-t farm granaries were filled with approximately 11 t of barley. The barley was either not treated, treated with protein-enriched pea flour at 0.1% throughout the entire grain mass, or treated at 0.5% throughout the top half of the grain mass. Adult insects were released in screened boxes (two insects per kilogram barley for S. oryzae and T. castaneum 1.4 insects per kilogram barley for C. ferrugineus). Barley was sampled four times during the 70-d trial. The number and mortality of adults and emerged adults in the samples were noted. Four kinds of traps, flight, surface-pitfall, probe-pitfall, and sticky-bar, were placed at different locations in the granaries to estimate the movement of insects. The 0.1% protein-enriched pea flour treatment reduced adult numbers of S. oryzae by 93%, T. castaneum by 66%, and C. ferrugineus by 58%, and reduced the emerged adults by 87, 77, and 77%, respectively. Treating the top half of the barley with 0.5% protein-enriched pea flour had similar effects as treating the entire grain mass with 0.1% pea-protein flour. However, the top-half treatment failed to prevent insects from penetrating into the untreated lower layer. Differences between traps are discussed.

Animals↗

A GDSL lipase confers resistance to piercing-sucking insects in tobacco by strengthening leaf cuticle.

Piercing-sucking insects, such as whiteflies and aphids, cause massive economic losses in major crops around the world. During feeding, the stylets of piercing-sucking insects navigate cuticles, cell walls, epidermal cells, and mesophyll cells; thus, these barriers are vital for the resistance of plants to insects. However, the relationship between insect stylet probing behavior and the composition and structure of these barriers remains unclear. Here, we identified a tobacco Cuticle Related Factor (NtCRF), which was induced significantly by whitefly infestation. Bioassays showed that NtCRF positively regulated plant resistance against whiteflies and green peach aphids. Silencing of NtCRF did not affect plant jasmonic acid (JA) and salicylic acid (SA) defenses but shortened the stylet probing time of phloem-feeders. Further studies confirmed that silencing of NtCRF resulted in significant structure destruction of the leaf cuticle and led to increased epidermal permeability. Overexpression of NtCRF in Arabidopsis also significantly enhanced the plant's resistance against whiteflies and green peach aphids. Our findings expand understanding of plant-insect interactions and provide a strategy for genetic improvement of crop resistance against piercing-sucking insects.

Animals↗

Induction of detoxication enzymes in insects.

Most of the known enzyme systems involved in the degradation of insecticides, other xenobiotics, and hormones of insects, appear to be inducible by various dietary chemicals, both plant-based and man-made. Although the understanding of the mechanism of this phenomenon comes almost entirely from studies of higher animals, there are good reasons to believe that the insect system is similar in most respects. If so, it involves a recognition system wherein exogenous chemicals in the insect's environment can stimulate the production of the enzymes that degrade them. Evidence is presented that some plant species and some plant allelochemicals are capable of such enhancement and that this is sufficient to increase the tolerance of an insect to various insecticides. It is also suggested that induction by plant chemicals may be involved in the insect's adaptation to its plant diet. Further understanding of the phenomenon, especially in regard to the cyclic nature of the process in plants and insects, may be of value in organizing pest management programs.

Adaptation, Physiological↗

The insect voltage-gated sodium channel as target of insecticides.

Examination of the function, chemistry, and pharmacology of the voltage-gated insect sodium channel (ISC) reveals that the ISC closely resembles its vertebrate counterpart in electrophysiology and ion conductance, primary structure and allocation of all functional domains, and its pharmacological diversity and flexibility exhibited by the occurrence of different allosterically coupled receptor-binding sites for various neurotoxicants. The toxicants include several groups of insecticides, namely DDT and its analogues, pyrethroids, N-alkylamides, and dihydropyrazoles, which affect channel gating and ion permeability. Despite their similarity, the insect and vertebrate channels are pharmacologically distinguishable, as revealed by the responsiveness of the heterologously expressed Drosophila para clone to channel modifiers and blockers and the occurrence of the insect-selective sodium channel neurotoxins derived from arachnid venoms presently used for the design of recombinant baculovirus-mediated selective bioinsecticides. The pharmacological specificity of the ISC may lead to the design of insect-selective toxicants, and its pharmacological flexibility may direct the use of ISC insecticides for resistance management. Insecticide resistance [such as knockdown resistance (KDR)] is acquired by natural selection and operated by increased metabolism, channel mutagenesis, or both. The resistance issue can be dealt with in several ways. One is by simultaneous application of low doses of synergistic, allosterically coupled mixtures (thus delaying or preventing the onset of resistance). An alternative is to replace an insecticide to which resistance was acquired by channel mutation with a different ISC toxicant to which increased susceptibility was conferred by the same mutation. Such a possibility was exemplified by a significant increase in susceptibility to N-alkylamides, as well as an insect-selective neurotoxin revealed by KDR insects. Third, both of these methods can be combined. Thus owing to its pharmacological uniqueness, the ISC may serve as a high-priority target for future selective and resistance-manageable insecticides.

Animals↗

Conservation of ecdysis-triggering hormone signalling in insects.

Pre-ecdysis- and ecdysis-triggering hormones (PETH and ETH) from endocrine Inka cells initiate ecdysis in moths and Drosophila through direct actions on the central nervous system (CNS). Using immunohistochemistry, we found Inka cells in representatives of all major insect orders. In most insects, Inka cells are numerous, small and scattered throughout the tracheal system. Only some higher holometabolous insects exhibit 8-9 pairs of large Inka cells attached to tracheae in each prothoracic and abdominal segment. The number and morphology of Inka cells can be very variable even in the same individuals or related insects, but all produce peptide hormones that are completely released at each ecdysis. Injection of tracheal extracts prepared from representatives of several insect orders induces pre-ecdysis and ecdysis behaviours in pharate larvae of Bombyx, indicating functional similarity of these peptides. We isolated several PETH-immunoreactive peptides from tracheal extracts of the cockroach Nauphoeta cinerea and the bug Pyrrhocoris apterus and identified the gene encoding two putative ETHs in the mosquito Anopheles gambiae. Inka cells also are stained with antisera to myomodulin, FMRFamide and other peptides sharing RXamide carboxyl termini. However, our enzyme immunoassays show that these antisera cross-react with PETH and ETH. Our results suggest that Inka cells of different insects produce only peptide hormones closely related to PETH and ETH, which are essential endocrine factors required for activation of the ecdysis behavioural sequence.

Animals↗

Safety of Angiotensin-converting enzyme inhibitors in patients with insect venom allergies.

OBJECTIVE: To review the literature with respect to the safety of angiotensin-converting enzyme (ACE) inhibitors in patients allergic to insect venom and those undergoing venom immunotherapy (VIT). DATA SOURCES: A MEDLINE search was conducted (1966-March 2006) using the following search terms: bee sting, venom, insect stings, ACE inhibitors, angiotensin II receptor blockers, immunotherapy, and desensitization. The bibliographies of qualifying articles were also searched for relevant references. DATA SYNTHESIS: Several case reports have described severe allergic reactions, including anaphylaxis, in patients taking ACE inhibitors subsequent to being stung or receiving VIT. Exacerbation of the allergic response by ACE inhibitors is thought to be related to accumulation of bradykinin and inhibition of the formation of angiotensin II. Similar reactions have not been described with angiotensin-receptor blockers, but are theoretically possible. CONCLUSIONS: ACE inhibitors may exacerbate the response to insect venom, resulting in potentially life-threatening allergic reactions to insect stings or VIT. Although this risk is difficult to quantify based only on data from case reports, it seems prudent that patients with documented allergic reactions to insect venom avoid ACE inhibitor therapy, if possible. If, after careful consideration of the risks and benefits, ACE inhibitor therapy is deemed warranted, education regarding measures to minimize exposure to insect stings and training on self-administration of epinephrine should be provided, as with any person with venom allergy. In patients in whom VIT is appropriate, temporary discontinuation of the ACE inhibitor prior to each venom injection may prevent subsequent adverse reactions.

Angiotensin-Converting Enzyme Inhibitors↗

Transformation systems in insects.

Genetic transformation is an important technology that provides unique opportunities to find, isolate, and analyze genes, as well as to create organisms with unique functional characteristics. Insect biologists have been developing genetic transformation technologies that rely extensively on transposable elements. A number of class II transposable elements isolated originally from insects have been converted into broad host range insect gene vectors. Class II transposable elements are particularly amenable to gene vector development, although they suffer from some limitations such as low rates of recombination. Use of these gene vectors requires the physical introduction of the vectors into developing insect embryos by microinjection. Microinjection methods vary to accommodate the unique physical and developmental characteristics of the target insects. All methods rely on the use of fine glass needles in conjunction with micromanipulators and a microscope. A serious constraint on the use of existing systems can be the inefficiency of successfully delivering the gene vectors to the germ cells of the developing embryo. The general method for vector delivery to insect germ cells is described, as well as variations that are useful under some conditions.

Animals↗

Eavesdropping on insects hidden in soil and interior structures of plants.

Accelerometer, electret microphone, and piezoelectric disk acoustic systems were evaluated for their potential to detect hidden insect infestations in soil and interior structures of plants. Coleopteran grubs (the scarabaeids Phyllophaga spp. and Cyclocephala spp.) and the curculionids Diaprepes abbreviatus (L.) and Otiorhynchus sulcatus (F.) weighing 50-300 mg were detected easily in the laboratory and in the field except under extremely windy or noisy conditions. Cephus cinctus Norton (Hymenoptera: Cephidae) larvae weighing 1-12 mg could be detected in small pots of wheat in the laboratory by taking moderate precautions to eliminate background noise. Insect sounds could be distinguished from background noises by differences in frequency and temporal patterns, but insects of similarly sized species could not be distinguished easily from each other. Insect activity was highly variable among individuals and species, although D. abbreviatus grubs tended to be more active than those of O. sulcatus. Tests were done to compare acoustically predicted infestations with the contents of soil samples taken at recording sites. Under laboratory or ideal field conditions, active insects within approximately 30 cm were identified with nearly 100% reliability. In field tests under adverse conditions, the reliability decreased to approximately 75%. These results indicate that acoustic systems with vibration sensors have considerable potential as activity monitors in the laboratory and as field tools for rapid, nondestructive scouting and mapping of soil insect populations.

Acoustics↗

Association of insects and ergot (Claviceps purpurea) in Kentucky bluegrass seed production fields.

Insects in Kentucky bluegrass seed production fields in Oregon, Idaho, and Washington were sampled just before harvest and their association with ergot conidia of Claviceps purpurea Fr. (Tul.) was evaluated during 1996-1998. A diversity of insects was observed at all three locations. The most abundant beneficial insects collected with sweep nets were Nysium spp., Nabis spp., ichneumonid wasps, and Hippodamia spp. The cranberry girdler, Chrysoteuchia topiaria (Zeller), was the only important pest on grass seed collected by sweep net. Numbers of aphids such as Sitobion avenae (F.), cicadellids and thrips such as Anaphothrips spp. and Aptinothrips spp. that were collected with all aphid sampler were below economic thresholds. Other insect groups occurred in low numbers. Noctuid moths collected in universal blacklight traps included nine species of cutworms and armyworms. Protogrotis obscura (B. & McD.) was the most common cutworm species and was present in all fields. The moth Chortodes rufostrigata (Pack.) previously reported only from wet meadows in northeast and south central Oregon was found in Kentucky bluegrass fields in central Oregon, suggesting that irrigated Kentucky bluegrass seed production fields may simulate a montane meadow habitat. Conidia of C. purpurea were found on a diversity of insects, including moths, flies, leafhoppers, and thrips. Up to 100% of moths and 75% of flies collected from some fields carried conidia of C. purpurea. No correlation between ergot honeydew present in a field and number of insects with conidia of C. purpurea was detected.

Animals↗

Transmission of Ehrlichia risticii, the agent of Potomac horse fever, using naturally infected aquatic insects and helminth vectors: preliminary report.

Ehrlichia risticii, the agent of Potomac horse fever (PHF), has been recently detected in trematode stages found in snail secretions and in aquatic insects. Based on these findings, horses could conceivably be exposed to E. risticii by skin penetration with infected cercariae, by ingestion of infected cercariae in water or via metacercariae in a second intermediate host, such as an aquatic insect. In order to test this hypothesis, horses were challenged with infectious snail secretions and aquatic insects collected from a PHF endemic region in northern California. Two horses stood with their front feet in water harbouring E. risticii-infected cercariae, 2 horses drank water harbouring E. risticii-infected cercariae, and 6 horses were fed pools of different aquatic insects harbouring E. risticii-infected metacercariae. In this preliminary study, only the one horse infected orally with mature caddisflies (Dicosmoecus gilvipes) developed the clinical and haematological disease syndrome of PHF. The agent was isolated from the blood of the infected horse in a continuous cell line and identified as E. risticii by characterisation of the 16S rRNA gene. Therefore, E. risticii is maintained in nature in a complex aquatic ecosystem and transmission to horses can occur through accidental ingestion of insects such as caddisflies containing infected metacercariae. At present, the small number of horses used in this study does not exclude other insects and free trematode stages as potential sources of infection.

Animals↗

Use of CD63 expression as marker of in vitro basophil activation in identifying the culprit in insect venom allergy.

BACKGROUND: The diagnosis of insect venom allergy and the indication for specific immunotherapy is based on history, skin tests and demonstration of hymenoptera venom specific IgE-antibodies. In cases with contradictory test results additional cellular tests are recommended. OBJECTIVE: We evaluated the usefulness of a newly introduced test based on basophil CD63 expression as marker of activation in comparison with the basophil histamine release test (BHR) and the cellular antigen stimulation test (CAST) measuring leukotriene release. METHODS: In 14 patients (10 males, 4 females; age; 12 to 67 years, mean: 42.5 +/- 15.1 years) with systemic reactions to hymenoptera stings in their history skin tests and determination of specific IgE-antibodies (CAP-RAST-FEIA) had shown inconsistent results: No demonstration of specific IgE-antibodies (n = 4), one sting by an unknown insect together with positive skin test and/or positive RAST to both bee and wasp venom (n = 4), several stings of partly unknown insects with positive skin test and/or demonstrable specific IgE-antibodies to more than one insect venom (n = 4), uncertain history and divergent results in skin test and/or RAST (n = 2). BHR, CAST and basophil activation test (BAT) were done according to the manufacturers with negative and positive controls and different concentrations of bee and wasp venom. The BAT is based on double staining with anti-IgE antibodies and anti-CD63 and subsequent determination of the percentage of activated basophils by flow cytometry. RESULTS: BAT and skin test were concordant in 42.9%, BAT and RAST in 57.1%. Concordance of all three cellular tests was seen in 57.1%, of BAT and BHR in 69.1%, of BAT and CAST in 78.6% and of BHR and CAST in 64.3%. In 6 cases where the three cellular tests (BHR, CAST, BAT) were not in accordance the addition of BAT led to a more reliable diagnostic result concerning the relevant insect in 3 cases and added no further information in 3 cases. BAT in controls always was negative. Correlation between CAST and BAT was higher than between CAST and BHR. CONCLUSIONS: In difficult cases of hymenoptera allergy, where history, skin tests and determination of specific antibodies do not allow a clear decision regarding the relevant insect species for immunotherapy, the additional performance of cellular tests (CAST and BAT) may be helpful.

Adolescent↗