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The effect of day of emergence from the insect cadaver on the behavior and environmental tolerances of infective juveniles of the entomopathogenic nematode Heterorhabditis megidis (strain UK211).

Infective juveniles (IJs) of entomopathogenic nematodes (EPNs) are obligate parasites of insect larvae. Inside the host they develop into sexually mature adult stages and complete their life cycle. Two or 3 adult nematode generations can occur in the insect host. The increase in nematode population density in the insect cadaver, together with limiting nutrient conditions, result in the formation of IJs. These IJs emerge into the soil to search for a new host. It typically takes 7-8 days for all IJs to emerge from a parasitized insect. We have investigated the effect of the day of emergence of IJs from insect cadavers on the environmental tolerance and behavior of the EPN Heterorhabditis megidis strain UK211. The IJs that emerge early display good initial host-finding ability and increased temperature tolerance but disperse poorly and have poor tolerance to desiccation. Conversely, the IJs that emerge later display poor initial host-finding ability and poor temperature tolerance but they disperse well and possess increased desiccation tolerance. These phenotypic differences are likely to facilitate early-emerging IJs in locating and infecting hosts in the vicinity of the cadaver, whereas IJs that emerge late are adapted to disperse away from their natal cadaver. We hypothesize that adaptive phenotypic plasticity rather than allelic variability may provide the genetic basis for the different physiological and behavioral phenotypes of the early- and late-emerging IJs.

Animals↗

The pattern of Distal-less expression in the mouthparts of crustaceans, myriapods and insects: new evidence for a gnathobasic mandible and the common origin of Mandibulata.

We examined embryos of representatives of crustaceans, myriapods and insects with respect to DII expression in the mouthparts. In order to examine the relationships between mandibular DII expression and the occurrence of a mandibular palp we compared amphipod, isopod and decapod crustacean species. In species with mandibular palps, DII expression is maintained throughout development and is restricted to the palps. The species lacking a palp as an adult show only transient DII expression in early embryonic stages. Furthermore, we studied mandibular DII expression in the myriapod Glomeris marginata that lacks like all myriapods mandibular palps as an adult. The expression pattern is similar to that in crustaceans lacking a palp as an adult. We examined entognathous and ectognathous insects. No sign of mandibular expression could be detected. It is shown that the distal parts of the mandibular appendage were reduced in several steps and lineages independently up to a total loss. Furthermore, we studied DII expression in the first and second maxillae. Except for Glomeris and the collembolans, the first maxillae of all species show a similar pattern of three lobes expressing DII: the outer expression marks the maxillary palp and the inner two mark the outgrowing endites (galea and lacinia of insects). In the first maxillae of collembolans only two expression areas could be detected. In palpless adult first maxillae of isopod crustaceans a transitory embryonic palp occurs which is also DII positive. In the second maxillae of insects, isopod and amphipod crustaceans only two DII-positive lobes occur. Our data suggest a gnathobasic character of the mandibles of crustaceans, myriapods and insects supporting the monophyly of Mandibulata sensu Snodgrass. The interpretation of DII expression patterns and its limits are critically evaluated.

Animals↗

Insect immunity: expression of the two major inducible antibacterial peptides, defensin and diptericin, in Phormia terranovae.

Injections of low doses of bacteria into larvae of Phormia terranovae induce the appearance of potent bactericidal peptides in the blood, among which predominate the anti-Gram positive insect defensins and the anti-Gram negative diptericins. Insect defensins show significant homologies to mammalian (including human) microbicidal peptides present in polymorphonuclear leukocytes and macrophages. We report the molecular cloning of cDNAs and primer extension studies which indicate that insect defensin is produced as a prepro-peptide yielding mature defensin A (40 residues) after cleavage of a putative signal peptide (23 residues) and a prosequence (34 residues). Previous studies have established that diptericin (82 residues) is matured from a pre-peptide by cleavage of a putative signal peptide (19 residues) and C-terminal amidation. Using oligonucleotide probes complementary to the sequences of the mRNAs for defensin and diptericin, we show by in situ hybridization that both antibacterial peptides are concomitantly synthesized by the same cells: thrombocytoids, a specialized blood cell type, and adipocytes. Transcriptional studies based on hybridization of RNAs to cDNAs of defensin and diptericin indicate that the transcription of both genes is induced regardless of the nature of the stimulus (injection of Gram positive or Gram negative bacteria, lipopolysaccharides). Even a sterile injury applied to axenically raised larvae is efficient in inducing the transcription of both genes suggesting that the local disruption of the integument aspecifically initiates a signalling mechanism which the thrombocytoids and the adipocytes are able to interpret. The transcription of immune genes is relatively short lived and a second challenge yields a response similar to that of the first stimulus, indicating that the experimental insects do not keep a 'memory' of their first injection.

Amino Acid Sequence↗

Biosynthesis and distribution of insect-molting hormones in plants--a review.

Insect-molting hormones, phytoecdysteroids, have been reported to occur in over 100 plant families. Plants, unlike insects, are capable of the biosynthesis of ecdysteroids from mevalonic acid, and in several cases the biosynthesis of phytoecdysteroids was also demonstrated to proceed via sterols. Spinacia oleracea (spinach) biosynthesizes polypodine B and 20-hydroxyecdysone, which is the predominant insect-molting hormone found in plant species. The onset of ecdysteroid production in spinach requires the appropriate ontogenetic development within the plant, which is related to leaf development. In spinach, lathosterol is the biosynthetic precursor to ecdysone and 20-hydroxyecdysone. Phosphorylated ecdysteroid intermediates, particularly ecdysone-3-phosphate, are required during biosynthesis. Polyphosphorylated forms of ecdysteroids are putative regulatory components of the pathway. During spinach development, the 20-hydroxyecdysone is transported from the sites of biosynthesis to the apical regions. An analysis of the physiological data available suggests that different species may synthesize ecdysteroids in various organs and distribute these ecdysteroids to other sites. Annual plants appear to concentrate ecdysteroids in the apical regions, including flowers and seeds. Perennial plants may recycle their ecdysteroids between their deciduous and their perennial organs over the growing season. Further investigations of ecdysteroid biosynthesis and physiology within plants will be required before an acceptable system can be designed to test phytoecdysteroid effectiveness in vivo against insect herbivory.

Ecdysteroids↗

Gastrointestinal digestion governs insect protein hydrolysis and predicted bioactive peptide release: Species-dependent implications for functional food applications.

This study investigates the digestion of insect proteins and the release of predicted bioactive peptides during human gastrointestinal digestion. Using the Infogest in vitro model, mealworm, cricket, and black soldier fly larvae (BSFL) proteins were digested and analyzed through discovery proteomics and bioinformatics to identify predicted bioactive peptides. Sequential windowed acquisition of all theoretical fragment ion mass spectra (SWATH-MS) quantified insect proteins including predicted bioactive peptide precursor proteins, the precursors of predicted bioactive peptides. Results indicated that gastrointestinal digestion strongly influences peptide release, with the gastric phase exhibiting a richer predicted bioactive peptide profile than the small intestinal phase. Many predicted bioactive peptides were rapidly hydrolysed under small intestine conditions, which may lead to reduced stability or diminished activity in vivo, potentially explaining why certain peptides show strong bioactivity in vitro but limited effects in vivo. Additionally, predicted bioactive peptide release varied by insect species, influenced by genetic factors and peptide abundance. These findings highlight the importance of species selection and consideration of proteolytic digestion patterns in optimizing insect-derived bioactive peptides for functional foods and nutraceutical applications.

Animals↗

Small RNA viruses of insects: expression in plants and RNA silencing.

Interest in insect small RNA viruses (SRVs) has grown slowly but steadily. A number of new viruses have been analyzed at the sequence level, adding to our knowledge of their diversity at the level of both individual virus species and families. In particular, a number of possible new virus families have emerged. This research has largely been driven by interest in their potential for pest control, as well as in their importance as the causal agents of disease in beneficial arthropods. At the same time, research into known viruses has made valuable contributions to our understanding of an emerging new field of central importance to molecular biology-the existence of RNA-based gene silencing, developmental control, and adaptive immune systems in eukaryotes. Subject to RNA-based adaptive immune responses in their hosts, viruses have evolved a variety of genes encoding proteins capable of suppressing the immune response. Such genes were first identified in plant viruses, but the first examples known from animal viruses were identified in insect RNA viruses. This chapter will address the diversity of insect SRVs, and attempts to harness their simplicity in the engineering of transgenic plants expressing viruses for resistance to insect pests. We also describe RNA interference and antiviral pathways identified in plants and animals, how they have led viruses to evolve genes capable of suppressing such adaptive immunity, and the problems presented by these pathways for the strategy of expressing viruses in transgenic plants. Approaches for countering these problems are also discussed.

Animals↗

Prolonged pheromonotropic activity of pseudopeptide mimics of insect pyrokinin neuropeptides after topical application or injection into a moth.

Amphiphilic pseudopeptide analogs of Phe-Thr-Pro-Arg-Leu-NH2, representing the active C-terminal core pentapeptide of the pyrokinin class of insect neuropeptides, were synthesized by replacement of phenylalanine with hydrocinnamic acid (Hca-Thr-Pro-Arg-Leu-NH2), or addition of 1-pyrenebutyric acid (Pba-Phe-Thr-Pro-Arg-Leu-NH2) or 9-fluoreneacetic acid (Fla-Phe-Thr-Pro-Arg-Leu-NH2). The pseudopeptides were found to stimulate sex pheromone biosynthesis when injected into females of the moth Heliothis virescens. Optimal pheromonotropic responses were obtained by injection of 0.25 pmol of Hca-Thr-Pro-Arg-Leu-NH2, 2.5 pmol of Pba-Thr-Pro-Arg-Leu-NH2 and 0.5 pmol of Fla-Thr-Pro-Arg-Leu-NH2. Topical application of each of the pseudopeptides in water to the cuticle of moths stimulated significant production of pheromone at a dose of 50 pmol with optimal stimulation occurring when 500 pmol were applied. The parent peptide, Phe-Thr-Pro-Arg-Leu-NH2, failed to stimulate significant production of pheromone when applied topically at a dose as high as 2000 pmol. Temporal studies indicated that Hca-Thr-Pro-Arg-Leu-NH2 stimulated significant production of pheromone for only 4 h after application where as continuous pheromone production for 18 h was observed when either Pba-Phe-Thr-Pro-Arg-Leu-NH2 or Fla-Phe-Thr-Pro-Arg-Leu-NH2 were applied to the abdomen. The results show that modification of the C-terminal active core of the insect pyrokinins, by addition of hydrophobic moieties, can result in production of pseudopeptides which effectively penetrate the insect cuticle and have prolonged physiological effects making them favorable candidates for use in development of alternative strategies for pest insect control.

Acetates↗

Antibody-mediated inhibition of the growth of larvae from an insect causing cutaneous myiasis in a mammalian host.

Many insects feed on blood or tissue from mammalian hosts. One potential strategy for the control of these insects is to vaccinate the host with antigens derived from the insect. The larvae of the fly Lucilia cuprina feed on ovine tissue and tissue fluids causing a cutaneous myiasis associated with considerable host morbidity and mortality. A candidate vaccine antigen, peritrophin 95, was purified from the peritrophic membrane, which lines the gut of these larvae. Serum from sheep vaccinated with peritrophin 95 inhibited growth of first-instar L. cuprina larvae that fed on this serum. Growth inhibition was probably caused by antibody-mediated blockage of the normally semipermeable peritrophic membrane and the subsequent development of an impervious layer of undefined composition on the gut lumen side of the peritrophic membrane that restricted access of nutrients to the larvae. The amino acid sequence of peritrophin 95 was determined by cloning the DNA complementary to its mRNA. The deduced amino acid sequence codes for a secreted protein containing a distinct Cys-rich domain of 317 amino acids followed by a mucin-like domain of 139 amino acids. The Cys-rich domain may be involved in binding chitin. This report describes a novel immunological strategy for the potential control of L. cuprina larvae that may have general application to the control of other insect pests.

Amino Acid Sequence↗

Iron metabolism in insects.

Like other organisms, insects must balance two properties of ionic iron, that of an essential nutrient and a potent toxin. Iron must be acquired to provide catalysis for oxidative metabolism, but it must be controlled to avoid destructive oxidative reactions. Insects have evolved distinctive forms of the serum iron transport protein, transferrin, and the storage protein, ferritin. These proteins may serve different functions in insects than they do in other organisms. A form of translational control of protein synthesis by iron in insects is similar to that of vertebrates. The Drosophila melanogaster genome contains many genes that may encode other proteins involved in iron metabolism.

Amino Acid Sequence↗

Insect odor and taste receptors.

Insect odor and taste receptors are highly sensitive detectors of food, mates, and oviposition sites. Following the identification of the first insect odor and taste receptors in Drosophila melanogaster, these receptors were identified in a number of other insects, including the malaria vector mosquito Anopheles gambiae; the silk moth, Bombyx mori; and the tobacco budworm, Heliothis virescens. The chemical specificities of many of the D. melanogaster receptors, as well as a few of the A. gambiae and B. mori receptors, have now been determined either by analysis of deletion mutants or by ectopic expression in in vivo or heterologous expression systems. Here we discuss recent advances in our understanding of the molecular and cellular basis of odor and taste coding in insects.

Animals↗

Pax group III genes and the evolution of insect pair-rule patterning.

Pair-rule genes were identified and named for their role in segmentation in embryos of the long germ insect Drosophila. Among short germ insects these genes exhibit variable expression patterns during segmentation and thus are likely to play divergent roles in this process. Understanding the details of this variation should shed light on the evolution of the genetic hierarchy responsible for segmentation in Drosophila and other insects. We have investigated the expression of homologs of the Drosophila Pax group III genes paired, gooseberry and gooseberry-neuro in short germ flour beetles and grasshoppers. During Drosophila embryogenesis, paired acts as one of several pair-rule genes that define the boundaries of future parasegments and segments, via the regulation of segment polarity genes such as gooseberry, which in turn regulates gooseberry-neuro, a gene expressed later in the developing nervous system. Using a crossreactive antibody, we show that the embryonic expression of Pax group III genes in both the flour beetle Tribolium and the grasshopper Schistocerca is remarkably similar to the pattern in Drosophila. We also show that two Pax group III genes, pairberry1 and pairberry2, are responsible for the observed protein pattern in grasshopper embryos. Both pairberry1 and pairberry2 are expressed in coincident stripes of a one-segment periodicity, in a manner reminiscent of Drosophila gooseberry and gooseberry-neuro. pairberry1, however, is also expressed in stripes of a two-segment periodicity before maturing into its segmental pattern. This early expression of pairberry1 is reminiscent of Drosophila paired and represents the first evidence for pair-rule patterning in short germ grasshoppers or any hemimetabolous insect.

Amino Acid Sequence↗

Spliceosomal small nuclear RNA genes in 11 insect genomes.

The removal of introns from the primary transcripts of protein-coding genes is accomplished by the spliceosome, a large macromolecular complex of which small nuclear RNAs (snRNAs) are crucial components. Following the recent sequencing of the honeybee (Apis mellifera) genome, we used various computational methods, ranging from sequence similarity search to RNA secondary structure prediction, to search for putative snRNA genes (including their promoters) and to examine their pattern of conservation among 11 available insect genomes (A. mellifera, Tribolium castaneum, Bombyx mori, Anopheles gambiae, Aedes aegypti, and six Drosophila species). We identified candidates for all nine spliceosomal snRNA genes in all the analyzed genomes. All the species contain a similar number of snRNA genes, with the exception of A. aegypti, whose genome contains more U1, U2, and U5 genes, and A. mellifera, whose genome contains fewer U2 and U5 genes. We found that snRNA genes are generally more closely related to homologs within the same genus than to those in other genera. Promoter regions for all spliceosomal snRNA genes within each insect species share similar sequence motifs that are likely to correspond to the PSEA (proximal sequence element A), the binding site for snRNA activating protein complex, but these promoter elements vary in sequence among the five insect families surveyed here. In contrast to the other insect species investigated, Dipteran genomes are characterized by a rapid evolution (or loss) of components of the U12 spliceosome and a striking loss of U12-type introns.

Animals↗

INSECT STING ANAPHYLAXIS.

Anaphylaxis from insect stings, which is considerably commoner than has been recognized, is a distinct emergency, requiring prompt and energetic treatment. Such reactions require the immediate intramuscular or deep hypodermic injection of 0.5 cc of 1:1000 epinephrine, which may have to be repeated shortly. After the initial critical phase is passed, there may be indication for intramuscular antihistamines and corticosteroids.Persons who have survived insect sting anaphylaxis should be immunized with insect antigens for a minimum period of three years and perhaps indefinitely. The choice between pure venom and extracts of whole insect bodies rests with the physician, although the latter are far more often used.Until immunization has become effective susceptible persons must carry with them at all times a kit containing epinephrine for both injection and aerosol use, and they must be trained by physicians in the proper use of these preparations.

Adrenal Cortex Hormones↗

Insect bite-like reaction in patients with hematologic malignant neoplasms.

BACKGROUND: Exaggerated reaction to insect bites, mainly to mosquitoes, is infrequently described in patients with chronic lymphocytic leukemia. Skin lesions usually appear months to years after the diagnosis of leukemia and are unrelated to laboratory findings, disease course, or therapy. OBSERVATIONS: We describe 8 patients with various hematologic disorders (chronic lymphocytic leukemia, acute lymphoblastic leukemia, acute monocytic leukemia, mantle-cell lymphoma, large-cell lymphoma, and myelofibrosis) who developed insect bite-like reaction. Although the clinical picture and the histological characteristics of the lesions were typical for insect bites, none of the patients actually had a history, course, or response to treatment suggestive of arthropod assaults. In 2 patients, the eruption preceded the diagnosis of the malignant neoplasm. The rash persisted for months to years and was resistant to therapies other than systemic corticosteroids. The 3 patients with chronic lymphocytic leukemia seemed to have a worse prognosis than expected for their disease. In 1, the polymerase chain reaction detected leukemic cells in the infiltrate. CONCLUSIONS: Insect bite-like reaction is an infrequent, disturbing, and difficult-to-treat nonspecific phenomenon in patients with hematologic malignant neoplasms. Since it may precede the hematologic disorder, oriented evaluation is warranted. We speculate that immunodeficiency plays a role in its pathogenesis; however, the exact pathogenesis and its prognostic implications await further studies.

Adolescent↗

Insect neuropeptide antagonists.

The development of a new integrated approach to the generation of a novel type of insect neuropeptide (Np) antagonists and putative insect control agents based on backbone cyclic compounds is described. The approach, termed the backbone cyclic neuropeptide-based antagonist (BBC-NBA), was applied to the insect pyrokinin (PK)/pheromone biosynthesis activating neuropeptide (PBAN) family as a model, and led to the discovery of a potent linear lead antagonist and several highly potent, metabolically stable BBC antagonists, devoid of agonistic activity, which inhibited PBAN-mediated activities in moths in vivo. This review briefly summarizes our knowledge of insect Nps, describes the PK/PBAN Np family, presents the basic concepts behind the BBC-NBA approach, and introduces the advantages of this method for generation of Np agonists, antagonists and insecticide prototype molecules.

Amino Acid Sequence↗

Coevolution of insect trypsins and inhibitors.

Many plant proteinase inhibitors have lysine at the P1 position, presumably to avoid hydrolysis by insect trypsins. Lepidopteran trypsins appear to have adapted to resist proteinase inhibitors through increased inhibitor hydrolysis and decreased binding to inhibitor hydrophilic reactive sites. Lepidopteran digestive trypsins prefer lysine at the P1 position and have substrate binding subsites more hydrophobic than trypsins from insects in other orders. All available sequences of sensitive and inhibitor-insensitive insect trypsins were aligned with porcine trypsin, for which interactions with Kunitz and Bowman-Birk inhibitor are known. After discounting conserved positions and positions not typical of sensitive or insensitive trypsins, the following residues were considered important to insect trypsin-PI interactions (chymotrypsin numbering): 60, 94, 97, 98, 99, 188, 190, 213, 215, 217, 219, 228. These residues support the Neighbor Joining analysis tree branches separating sensitive and insensitive trypsin sequences. Primary sequences interacting with PIs are around the active site, with some forming part of the S1 (188, 217, 219 and 228) or S4 (99, 215) pockets.

Amino Acid Sequence↗

The influence of fluorinated molecules (semiochemicals and enzyme substrate analogues) on the insect communication system.

Can the introduction of fluorine atoms affect the bioactivity of natural semiochemicals? Can fluorine contribute in the creation of specific enzyme inhibitors to interrupt or disrupt the insect communication system? The first step for the bioactivity of a molecule is interaction with the biological sensor. Hydrogen and fluorine are almost bioisosteric and the receptor site of the enzyme can still recognize and accept the fluoro analogue of its natural substrate. However, the peculiar electronegativity of the fluorine atom can affect the binding, absorption, and transport of the molecule. The differences in the molecule's electronic properties can lead to differences in the chemical interactions between the receptor and the fluorinated substrate. Fluorine introduction can modify the metabolic stability and pathway of the semiochemicals in many different ways. Fluorinated analogues can show synergism, inhibition, or hyperagonism effects on insect behaviors, that is, the activity of the nonfluorinated parent compounds can be mimicked, lost, or increased. In any case, the fluorinated molecules can interact with the bioreceptors in a new and disrupting way. The semiochemicals are olfactory substances: fluorine can affect their volatility or smell. Production of semiochemicals from exogenous substances, perception at antennal receptors, and processing of biological responses are the main steps of communication among insects. In the production step, the fluorinated molecules can interact with enzymes that catalyze the biosynthesis of the natural pheromones. In the perception step, fluorinated semiochemicals can interact with the olfactory receptor cells; this often leads to totally unpredictable behaviors. Fluorinated molecules have been developed as probes to elucidate the complex chemorecognition processes of insects. Many of these molecules have been tested to find highly effective behavior-modifying chemicals. New analogues have been synthesized to investigate the metabolic pathway of a pheromone molecule and many of them are promising disrupting agents. Despite such titanic research efforts, the results have often been random, rational trends in the induced behaviors have sometimes been impossible to find, and practical applications of the fluorinated semiochemicals are still uncertain.

Alcohols↗