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The gut bacteria of insects: nonpathogenic interactions.

The diversity of the Insecta is reflected in the large and varied microbial communities inhabiting the gut. Studies, particularly with termites and cockroaches, have focused on the nutritional contributions of gut bacteria in insects living on suboptimal diets. The indigenous gut bacteria, however, also play a role in withstanding the colonization of the gut by non-indigenous species including pathogens. Gut bacterial consortia adapt by the transfer of plasmids and transconjugation between bacterial strains, and some insect species provide ideal conditions for bacterial conjugation, which suggests that the gut is a "hot spot" for gene transfer. Genomic analysis provides new avenues for the study of the gut microbial community and will reveal the molecular foundations of the relationships between the insect and its microbiome. In this review the intestinal bacteria is discussed in the context of developing our understanding of symbiotic relationships, of multitrophic interactions between insects and plant or animal host, and in developing new strategies for controlling insect pests.

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The antennal lobe of orthoptera - anatomy and evolution.

The first odor-processing neuropils of insects comprise glomeruli, islets of neuropil, that are supplied by olfactory receptor neurons and give rise to efferent axons to higher brain centers. Glomeruli size and organization varies in a taxon-specific manner across the Insecta, suggesting possible correlates between their organization and chemosensory behaviors in different insect groups. Comparative studies of antennal lobe glomeruli within the Orthoptera have been used to infer how the various taxon-specific arrangements of odorant-processing structures (glomeruli) might have evolved. The cellular arrangements in glomeruli have been surveyed using anterograde filling and Golgi impregnation of antennal receptor neurons projecting to the antennal lobe in Stenopelmatidae, Tettigoniidae, Gryllidae, Tetrigidae and Acrididae. These taxa, which represent the two sub-orders of Orthoptera, reveal a high correlation between the neural architecture of the glomeruli and structures within the glomeruli. Using a recent molecular phylogeny of the Orthoptera we have mapped the occurrence of glomerular characteristics to infer the evolution of antennal lobe structures in orthopterans. The functional implications of these results are discussed.

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The evolution of insect flight: implications for the evolution of the nervous system.

The Insecta encompasses a prodigiously diverse group as measured at the species, family and ordinal levels, but the nervous system bears evidence of conservatism. The early acquisition of flight must have been a major factor in the diversification of body form. Arguments are presented that predator evasion was a primary factor in the origin of flight and that a conserved set of giant interneurons played a key element in the transition.

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Distribution of PER protein, pigment-dispersing hormone, prothoracicotropic hormone, and eclosion hormone in the cephalic nervous system of insects.

Investigations performed on adult insects revealed that putative components of the central pacemaker, the protein Period (PER) and the pigment-dispersing hormone (PDH), are immunocytochemically detectable in discrete sets of brain neurons throughout the class of Insecta, represented by a bristletail, mayfly, damselfly, 2 locust species, stonefly, 2 bug species, goldsmith beetle, caddisfly, honeybee, and 2 blowfly species. The PER-positive cells are localized in the frontal protocerebrum and in most species also in the optic lobes, which are their only location in damselfly and goldsmith beetle. Additional PER-positive cells occur in a few species either in the deuto- and tritocerebrum or in the suboesophageal ganglion. The PER staining was always confined to the cytoplasm. The PDH immunoreactivity consistently occurs in a cluster of perikarya located frontoventrally at the proximal edge of the medulla. The mayfly and both locust species possess additional PDH neurons in 2 posterior cell clusters at the proximal edge of the medulla, and mayfly, waterstrider, and 1 of the blowfly species in the central brain. PDH-positive fibers form a fanlike arrangement over the frontal side of the medulla. Two or just 1 bundle of PDH-positive fibers run from the optic lobe to the protocerebrum, with collaterals passing over to the contralateral optic lobe. Antisera to the prothoracicotropic (PTTH) and the eclosion (EH) hormones, which in some insects regulate the molting and ecdysis rhythms, respectively, typically react with a few neurons in the frontal protocerebrum. However, the PTTH-positive neurons of the mayfly and the damselfly and the EH-positive neurons of the caddisfly are located in the suboesophageal ganglion. No PTTH-like antigen was detected in locusts, and no EH-like antigens were detected in the damselfly, stonefly, locusts, and the honeybee. There are no signs of co-localization of the PER-, PDH-, PTTH-, and EH-like antigens in identical neurons.

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Insect gas exchange patterns: a phylogenetic perspective.

Most investigations of insect gas exchange patterns and the hypotheses proposed to account for their evolution have been based either on small-scale, manipulative experiments, or comparisons of a few closely related species. Despite their potential utility, no explicit, phylogeny-based, broad-scale comparative studies of the evolution of gas exchange in insects have been undertaken. This may be due partly to the preponderance of information for the endopterygotes, and its scarcity for the apterygotes and exopterygotes. Here we undertake such a broad-scale study. Information on gas exchange patterns for the large majority of insects examined to date (eight orders, 99 species) is compiled, and new information on 19 exemplar species from a further ten orders, not previously represented in the literature (Archaeognatha, Zygentoma, Ephemeroptera, Odonata, Mantodea, Mantophasmatodea, Phasmatodea, Dermaptera, Neuroptera, Trichoptera), is provided. These data are then used in a formal, phylogeny-based parsimony analysis of the evolution of gas exchange patterns at the order level. Cyclic gas exchange is likely to be the ancestral gas exchange pattern at rest (recognizing that active individuals typically show continuous gas exchange), and discontinuous gas exchange probably originated independently a minimum of five times in the Insecta.

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Unusually long palindromes are abundant in mitochondrial control regions of insects and nematodes.

BACKGROUND: Palindromes are known to be involved in a variety of biological processes. In the present investigation we carried out a comprehensive analysis of palindromes in the mitochondrial control regions (CRs) of several animal groups to study their frequency, distribution and architecture to gain insights into the origin of replication of mtDNA. METHODOLOGY/PRINCIPAL FINDINGS: Many species of Arthropoda, Nematoda, Mollusca and Annelida harbor palindromes and inverted repeats (IRs) in their CRs. Lower animals like cnidarians and higher animal groups like chordates are almost devoid of palindromes and IRs. The study revealed that palindrome occurrence is positively correlated with the AT content of CRs, and that IRs are likely to give rise to longer palindromes. CONCLUSIONS/SIGNIFICANCE: The present study attempts to explain possible reasons and gives in silico evidence for absence of palindromes and IRs from CR of vertebrate mtDNA and acquisition and retention of the same in insects. Study of CRs of different animal phyla uncovered unique architecture of this locus, be it high abundance of long palindromes and IRs in CRs of Insecta and Nematoda, or short IRs of 10-20 nucleotides with a spacer region of 12-14 bases in subphylum Chelicerata, or nearly complete of absence of any long palindromes and IRs in Vertebrata, Cnidaria and Echinodermata.

AT Rich Sequence↗

Antihemostatic strategies of blood-feeding arthropods.

Arthropods in at least 23 different families or orders, distributed between two classes (Insecta and Arachnida), feed on vertebrate blood. They are able to do this despite constraints imposed by a sophisticated array of hemostatic defenses, due to the presence of a wide range of antihemostatic molecules in their saliva, including vasodilators, antiplatelet factors, and anticoagulants. Vasodilators include amines, prostaglandins, peptides, proteins, and even a mechanism to store large amounts of nitric oxide and deliver it into the skin. Platelet aggregation inhibitors include nitric oxide, prostaglandins, apyrase, molecules that sequester ADP, and a range of peptides and proteins that interact specifically with integrin receptors. Anticoagulants include a wide variety of inhibitors that target thrombin and factor Xa, as well as proteins that disrupt the "tenase", prothrombinase, and tissue factor/FVIIa complexes. The potential complexity of saliva is illustrated with the example of Rhodnius prolixus, which contains a large array of compounds, many of which affect more than one target in the hemostatic process. Finally a brief discussion of a new approach (sialomics) to the discovery of pharmacological agents in arthropod saliva is presented.

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Comparative biochemical, histochemical and autoradiographic studies of Na+/k+-ATPase in the rectum of dragonfly larvae (Odonata, Aeshnidae).

Na+/K+-ATPase localization in the rectal wall of larval Aeshna cyanea (Insecta, Odonata) was studied with histochemical precipitation techniques and 3H-ouabain autoradiography in conjunction with biochemical measurements of enzyme activities and radiospectrometry of 3H-ouabain binding, respectively. The NPP-strontium and ATP-lead methods led to complete inhibition of Na+/K+-ATPase in this organ and hence to unreliable histochemical results. The 3H-ouabain binding technique revealed sodium pump sites at the basolateral plasma membranes of the absorptive rectal chloride epithelia.

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[Gregarines from South Korea (author's transl)].

Study of 14 species of Gregarines from terrestrial arthropods (Myriapoda and Insecta) of south Korea. Some of them (Ramicephalus ozakii, Gregarina monoducta, Hoplorhynchus ozakii, Stylocephalus bahli) are typically asiatic whereas the others were already described from identical or similar european hosts.

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[Morphometric invariants of plants, insects and vertebrates].

The morphometric invariant construction is considered. The invariants are the numbers generated in an axiomatic way. Nonempirical invariants coincide with the empirical ones of various species and are calculated by three functions of two independent variables that are natural numbers. The first function is a quantitative description of well-known spectra of empirical plant invariants in the form of phyllotaxis ratios. The second and the third functions are generated from the first one. They describe quantitatively Insecta and Vertebrata empirical morphometric invariants in the form of double ratio among three linear sizes.

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Distribution of GABA-like immunoreactive neurons in insects suggests lineage homology.

Gamma-aminobutyric acid (GABA) is an important inhibitory neurotransmitter in vertebrates and invertebrates (Sattelle [1990] Adv. Insect Physiol. 22:1-113). The GABA phenotype is lineally determined in postembryonic neurons in the tobacco hawkmoth, Manduca sexta (Witten and Truman, [1991] J. Neurosci. 11:1980-1989) and is restricted to six identifiable postembryonic lineages in the moth's thoracic hemiganglia. We used a comparative approach to determine whether this distinct clustering of GABAergic neurons is conserved in Insecta. In the nine orders of insects surveyed (Thysanura, Odonata, Orthoptera, Isoptera, Hemiptera, Coleoptera, Diptera, Lepidoptera, and Hymenoptera), GABA-like immunoreactive neurons within a thoracic hemiganglion were clustered into six distinct groups that occupied positions similar to the six postembryonic lineages in Manduca. On the basis of cell body position and axon trajectories, we suggest that these are indeed homologous lineage groups and that the lineal origins of the GABAergic cells have been very conservative through insect evolution. The distinctive clustering of GABA-positive cells is shared with crustaceans (Mulloney and Hall [1990] J. Comp. Neurol. 291:383-394; Homberg et al. [1993] Cell Tissue Res. 271:279-288) but is not found in the centipede Lithobius forficulatus. There is a two- to threefold increase in numbers of thoracic neurons between the flightless Thysanura and the most advanced orders of insects. Using the GABA clusters as indicators of specific lineages, we find that only selected lineages have significantly contributed to this increase in neuronal numbers.

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Distribution of histamine in the CNS of different spiders.

Immunohistochemistry is used to demonstrate histamine-immunoreactivity in the CNS of spiders. We found histamine-immunoreactivity in the photoreceptors of different spiders. Therefore, we suggest that histamine is a neurotransmitter of photoreceptors in all arthropods, since it is also known to occur in the photoreceptors of the other main arthropod taxa (Merostomata, Crustacea, and Insecta). We also describe a system of only six omnisegmental histamine-immunoreactive neurons within the central nervous system. These histamine-immunoreactive neurons can be divided into two subgroups: a dorsal system with two cells per hemisphere and a ventral system with only one cell per hemisphere. All six cells have extended arborizations in both the motor and the sensory areas of all neuromeres in the suboesophageal ganglionic mass. In contrast to araneomorph spiders, two additional sets of histamine-immunoreactive neurons were detected in mygalomorph spiders. The first set consists of seventeen cells with their cell bodies located in the cheliceral ganglion and projecting to central areas of the protocerebrum. The second set contains many if not all sensory projections from the tarsal organs on all eight legs and the pedipalps to the Blumenthal neuropil.

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Identification and molecular analysis of storage proteins from Heliothis virescens.

Three abundant storage proteins have been detected in larval and pupal hemolymph and pupal fat body of the tobacco budworm, Heliothis virescens. These polypeptides have subunit molecular weights of 74,000, 76,000, and 82,000, as determined by SDS-PAGE and exist as 450,000-Mr hexamers in their native state. A purified 82,000-Mr storage protein fraction has been obtained along with a preparation containing equivalent amounts of the 74,000-Mr and 76,000-Mr subunits, and antisera raised to each of these components have been used to document the developmental profiles of protein accumulation and synthesis by fat body. cDNA clones corresponding to each of three abundant classes of fat body mRNAs have been recovered, and at least one of these has been unambiguously demonstrated to encode the 82,000-Mr storage protein subunit. Northern blot studies with these cDNA clones revealed that the developmental accumulation of transcripts in fat body for each was consistent with the general pattern of storage protein biosynthesis, and more interestingly, that transcripts hybridizing to two of these cDNA sequences are also found in tests. These two cDNAs have also been sequenced revealing that one encodes a polypeptide similar to arylphorins, a class of storage proteins widely distributed in Insecta. The derived amino sequence of the second cDNA, corresponding to the 82,000-Mr protein, had no unusual compositional features and determination of its structural relationship to other hemolymph polypeptides awaits molecular analysis of related genes from other insects.

Amino Acid Sequence↗

Innervation is necessary for the development of fast contraction kinetics of singing muscles in a katydid.

The twitch duration of mesothoracic wing muscles of the male katydid Neoconocephalus robustus (Insecta; Orthoptera; Tettigoniidae) decreases rapidly within the first 5 days of adulthood, to about half of its value in newly molted adults. To determine if this change is dependent upon neural input, male mesothoracic first tergocoxal muscles were unilaterally denervated on the second day of adulthood. The contraction kinetics of the denervated and contralateral innervated muscles were tested four days later. The development of rapid contraction kinetics was slowed or stopped in the denervated muscles, while the contralateral innervated muscles did become faster. Mesothoracic wing muscles of females do not develop faster contraction kinetics. When the female mesothoracic first tergocoxal muscle is denervated, there is no difference in twitch duration after 4 days between the innervated and contralateral denervated muscles. Therefore, denervation in newly molted adult male katydids interrupts a developmental program for the acquisition of adult contraction kinetics.

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Quantification and ultrastructure of oosorption in Eretmocerus eremicus (Hymenoptera: Aphelinidae).

The potential for and ultrastructure of oosorption were examined in Eretmocerus eremicus, a short-lived whitefly parasitoid that obligately produces anhydropic oocytes. In the absence of hosts, median egg load decreased by approximately 12% per day between 2 days and 8 days following eclosion. Parasitoid mating status had no significant effect on either egg load alone or the relationship between egg load and age. Yolk degradation in E. eremicus is autolytic, with the enzymes required for yolk sphere digestion apparently being derived from within the ooplasm. The exochorion appear to be digested by the follicular epithelium concurrent with the uniform degradation of the entire ooplasm. The potential adaptive benefits of this novel oosorption mechanism to E. eremicus females include a reduction in the total digestion time per oocyte and resorption of chorion remnants. Finally, to our knowledge, the results of this study provide the first unequivocal ultrastructural evidence of a purely autolytic oosorption mechanism in Insecta.

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Conservation of antigen 3G6: a crystalline cone constituent in the compound eye of arthropods.

A monoclonal antibody (MAb), 3G6, highly selective for neuropil glia in the CNS of the house cricket Acheta domesticus, also demonstrates remarkable selectivity for the nonneuronal crystalline cone cells of the compound eye. MAb 3G6 labels cone cells in eucone eyes throughout Insecta, from ancestral forms such as the bristle tail to the more recent honeybee; eucone structures are also recognized in Crustacea. Analogous nonneural structures found in pseudocone or acone eyes also express detectable 3G6 immunoreactivity. Immunoblot analysis demonstrates that MAb 3G6 binds to similar Mr85 kDa glycoproteins in the cricket CNS and retina, corresponding to the glial and crystalline cone forms of the antigen. Further, polypeptides of similar relative mass are also recognized in the eucone eye of the butterfly Pieris and the pseudocone eye of the fly Calliphora. The properties and function of glycoprotein 3G6 in the CNS and retina are yet to be explored. However, the finding that a unique antigen is highly conserved within the crystalline cone or analogous regions of the retina throughout the Arthropoda lends support at the molecular level to the notion that the arthropod compound eye has a monophyletic origin.

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Purification of toxic compounds from larvae of the gray fleshfly: the identification of paralysins.

Larval haemolymph of Neobellieria bullata (Insecta, Diptera) is highly toxic to adults of the same species: injection causes instant paralysis to death. Referring to their dramatic effect in adult insects the responsible compounds were designated paralysins. Two paralysins, soluble in organic solvents and heat stable, were chromatographically purified to homogeneity. They were identified by use of mass spectrometry and nuclear magnetic resonance respectively as beta-alanine-tyrosine (beta-Ala-Tyr) and as 3-hydroxy-kynurenine (3-HK). The quantities of beta-Ala-Tyr and 3-HK in the insect appear to increase steadily during larval development, with peak values prior to the pupal stage. These findings may contribute to a better understanding of some aspects of the process of insect metamorphosis. Orienting experiments in mammals suggest that both compounds, when injected intraspinally, are also neurotoxic to rats. In addition, cytotoxicity tests revealed that 3-HK, but not beta-Ala-Tyr is toxic to human neuroblastoma cells, rat primary cortex neurons as well as to rat glial cells.

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Evidence for growth of strains of the plant epiphytic bacterium Erwinia herbicola and transconjugation among the bacterial strains in guts of the silkworm Bombyx mori.

Growth of plant epiphytic bacteria Erwinia herbicola and Pseudomonas syringae in guts of the silkworm, Bombyx mori, was studied. Fifth instar silkworm larvae were fed artificial diets supplemented with these bacteria for 6 to 12 h followed by uncontaminated diets. At 1, 3, and 6 days after feeding, bacteria were isolated from insect guts and feces. A much larger population of E. herbicola was detected in the samples collected 3 and 6 days after the inoculation than in samples collected after 1 day, indicating that these bacteria grew in the insect gut, while P. syringae was unable to survive. Transconjugation between E. herbicola strains in the insect gut was also examined. First, either a donor or a recipient strain was fed to the insects in artificial diets containing the bacteria during 12 h, and then pairing strains were fed during 12 h after starvation for 12 h. The conjugative plasmid pBPW1::Tn7 was transferred into recipient cells at very high frequencies (10(-1)/recipient after 3 days and 10(-3) after 6 days) in insect guts. Indigenous plasmids of E. herbicola mobilized RSF1010 plasmid into recipient cells at frequencies of 10(-4) in insect guts. These transconjugants were detected in the feces of the insects. Thus, plasmid-mediated gene transfer among the epiphytic bacteria in insect guts was demonstrated. The results obtained suggest that in insecta gene transfer may play an important role in the evolution of plant epiphytic bacteria.

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