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Evolutionary history of Stratiomyidae (Insecta: Diptera): the molecular phylogeny of a diverse family of flies.

Stratiomyidae is a cosmopolitan family of Brachycera (Diptera) that contains over 2800 species. This study focused on the relationships of members of the subfamily Clitellariinae, which has had a complicated taxonomic history. To investigate the monophyly of the Clitellariinae, the relationships of its genera, and the ages of Stratiomyidae lineages, representatives for all 12 subfamilies of Stratiomyidae, totaling 68 taxa, were included in a phylogenetic reconstruction. A Xylomyidae representative, Solva sp., was used as an outgroup. Sequences of EF-1alpha and 28S rRNA genes were analyzed under maximum parsimony with bootstrapping, and Bayesian methods to recover the best estimate of phylogeny. A chronogram with estimated dates for all nodes in the phylogeny was generated with the program, r8s, and divergence dates and confidence intervals were further explored with the program, multidivtime. All subfamilies of Stratiomyidae with more than one representative were found to be monophyletic, except for Stratiomyinae and Clitellariinae. Clitellariinae were distributed among five separate clades in the phylogeny, and Raphiocerinae were nested within Stratiomyinae. Dating analysis suggested an early Cretaceous origin for the common ancestor of extant Stratiomyidae, and a radiation of several major Stratiomyidae lineages in the Late Cretaceous.

Animals↗

Evolution of host specialization in the Adelgidae (Insecta: Hemiptera) inferred from molecular phylogenetics.

The Adelgidae form a small group of insects in the Aphidoidea. They are cyclically parthenogenetic with host alternating, multiple-generation complex life cycles and are restricted to certain host genera in the Pinaceae. Species that host alternate always have Picea as the primary host where sexual reproduction and gall formation occur, and another genus in the Pinaceae as the secondary host where a series of parthenogenetic generations are produced. Other species that do not host alternate complete their entire life cycle on one host and only reproduce parthenogenetically. We studied relationships within Adelgidae using DNA sequences from the mitochondrial COI, COII, and cytb genes, and the nuclear EF1alpha gene. Analysis of the combined data resulted in a well-resolved phylogeny in which the major adelgid clades correspond neatly to their association with secondary host genera. Specialization on each secondary host genus occurred only once and was followed by diversification on the host genus. Molecular dating of divergence times in the Adelgidae suggest that diversification among host genera occurred in the Late Cretaceous and Early Tertiary when the Pinaceae genera were diverging. It is not clear, however, whether the Adelgidae and Pinaceae co-diversified because the relationships among the Pinaceae genera are not fully resolved. We discuss implications for adelgid taxonomy, life cycle evolution, and evolution of the interaction between adelgids and their host plants.

Animals↗

Studies on the toxicity of cypermethrin and fenvalerate in different strains of Drosophila melanogaster Meig. (Insecta, Diptera).

To improve our knowledge of pyrethroid toxicity mechanisms, Drosophila melanogaster was chosen as a model species, with well-defined characteristics. This study reports the results of the toxic effects of cypermethrin and fenvalerate on larvae and adults of four D. melanogaster strains. Our results show that cypermethrin is more toxic than fenvalerate at larval and adult stages, that growth on media containing insecticide significantly decreases the rate of preimaginal development, and that adult males are more sensitive than are females to both insecticides.

Animals↗

Immune suppression of Galleria mellonella (Insecta, Lepidoptera) humoral defenses induced by Steinernema feltiae (Nematoda, Rhabditida): involvement of the parasite cuticle.

Immune depression of Galleria mellonella larvae was evaluated a short time after infection with the entomopathogenic nematode Steinernema feltiae. In the host the activity of the enzymatic cascade known as the proPO system was significantly reduced by the presence of either live or dead parasites. The presence of parasites decreased the LPS-elicited proPO system activity. In addition, this process seems to be related to a decrease in the activity of hemolymph proteases, more than to phenoloxidase damage. proPO inhibition was also achieved by injected isolated cuticle fragments, suggesting that the parasite body surface plays an important role in the early parasitation phase.

Animals↗

The effect of azadirachtin on Blastocrithidia triatomae and Trypanosoma cruzi in Triatoma infestans (Insecta, Hemiptera).

The effect of azadirachtin on Blastocrithidia triatomae and Trypanosoma cruzi, which colonise the intestinal tract of the blood-sucking bug Triatoma infestans, was investigated. In established infections of controls without azadirachtin treatment, the small intestine of fifth-instar T. infestans contained up to 7 x l0(6) B. triatomae and the rectum 3 x 10(6). In comparison to this homoxenous flagellate, the population densities of T. cruzi in the respective regions were 99.3 and 76% lower. Treatment with azadirachtin (1 microg ml(-1)) via a blood meal and a concurrent infection with B. triatomae resulted in an increase of the population density (3 weeks p.i.), caused mainly by the mastigote stages in the rectum. In an established B. triatomae infection (12 weeks p.i.), feeding of azadirachtin did not affect the population density and composition. In an optimal T. cruzi-vector system, i.e. parasite and bug originate from the same locality, the treatment with azadirachtin at 20 weeks p.i. strongly reduced the population density in the small intestine of all bugs up to 100 days after treatment, but only in a minor percentage of the bugs in the rectum. Trypanosoma (cruzi incubated for up to 24 h in faeces of azadirachtin-treated bugs were not affected, indicating that the rectum of these bugs contained no toxic substances. The importance of these findings is that investigations of the mechanisms of action of azadirachtin offer a possibility to identify vector-derived compounds, which are necessary for the development of T. cruzi, thereby, giving us a possible new strategy to combat Chagas' disease.

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On the respiratory quotient (RQ) of termites (Insecta: Isoptera).

The respiratory quotient (RQ) at 28 degrees C was determined by Warburg manometry in 23 species of termites from the Mbalmayo Forest Reserve (Cameroon) and three sub-tropical species cultured under laboratory conditions in the U.K. or freshly collected in Australia. The data are tabulated with other recently reported RQs (determined by manometry or GC) and with measured CH(4) emission rates to provide a survey of 29 species covering both lower and higher termites in all major trophic (functional) categories. In all species, except the wood-feeding Coptotermes acinaciformis and the soil-feeding Cubitermes fungifaber, the observed mean values (with manometry corrected for known fluxes of H(2) and CH(4)) were at or well above 1.00. Soil-feeding forms (except C. fungifaber) generally showed a high apparent RQ (not corrected for H(2)), with nine species (out of 13) above 1.20 and six species above 1.30. Well-replicated laboratory experiments with Reticulitermes lucifugus showed that there was a tendency for RQ to fall with time over a 4-h incubation, although remaining greater than 1.00.The observed RQs are consistent with carbohydrate being the principal substrate supporting respiration in all trophic and taxonomic categories, with little or no contribution from the degradation of lignin or other polyaromatic materials. However, in many species (especially soil-feeders), the observed RQ is greater than that expected from known fluxes of O(2), CO(2) and CH(4) on the assumption that carbohydrate is the respiratory substrate. This presupposes that there is a large hydrogen sink (additional to CH(4) production), possibly the emission of H(2) gas, and/or the existence of unresolved digestive mechanisms or electron routings. Uncertainties in the use of manometry with termites are discussed.

Journal Article↗

Origin and pathway of the epidermal secretion in the damselfly head-arresting system (Insecta: Odonata).

In damselflies, the arrester system is responsible for an additional attachment of the head to the neck. It consists of a pair of mobile postcervical sclerites (SPC) covered by microtrichia. In their lateral position, SPCs can fixate the head on fields of microtrichia on the back surface of the head. The intact surface of microtrichia of the SPC is usually covered by a lipid-containing secretion. The present study provides ultrastructural data on the secretory epidermis and pore channels adapted to transport the secretion to the cuticle surface. 1) Shock-frozen preparations of the contact area show that microtrichia of co-opted surfaces do not interlock with each other. When co-opted fields are pressed to each other, deformations of flexible microtrichia of the SPC result in an increase of contact area between corresponding structures and consequently in an increase of frictional forces. 2) In the area of the SPC, only electron-lucent vesicles have been found. Histochemical procedures revealed that the material stored in vesicles and liberated on the external surface of the SPC is presumably non-volatile lipid. 3) Only a small number of large pore channels reach the surface of a microtrichium. Most of them terminate with tiny terminal channels, whose diameter is approximately six to twenty times smaller than the diameter of structures of secretory blooms occurring in SEM in shock-frozen and air-dried preparations. The terminal channels do not penetrate the epicuticular layer. It seems that the secretion reaches the epicuticle through terminal channels and diffuses through the epicuticle without any channel structures.

Journal Article↗

Differences between the pathogenic processes induced by Steinernema and Heterorhabditis (Nemata: Rhabditida) in Pseudaletia unipuncta (Insecta: Lepidoptera).

Larvae of Pseudaletia unipuncta are moderately susceptible to infections caused by entomopathogenic nematodes, being a desirable host to study pathogenic processes caused by Heterorhabditis bacteriophora, Steinernema carpocapsae, and Steinernema glaseri and their associated bacteria. The ability of the infective stage of these nematodes to invade hosts is quite different. S. carpocapsae invades the highest number of insects and presents the highest penetration rate, followed by H. bacteriophora. Regression analysis between the number of insects parasitized and the number of IJs counted per insect, over time, showed a high correlation for S. carpocapsae whereas for H. bacteriophora it was low. Dose-response was most evident at a concentration below 100 IJs per insect on H. bacteriophora, whereas on S. carpocapsae it was found for doses ranging from 100 to 2,000 IJs. Student's t test analysis of dose-response showed parallel, yet unequal, slopes for both strains of H. bacteriophora, whereas distinct regressions were obtained for S. carpocapsae and S. glaseri, thus, evidencing each species develop a distinct pathogenic process. Insects injected with Photorhabdus luminescens died within 50 h after injection, whereas those treated with X. nematophila died much later. Moreover, the mortality in insects exposed to H. bacteriophora complex and injected with P. luminescens was close, but insects injected with bacteria died faster. Insect mortality in treatments with complexes S. carpocapsae and S. glaseri was significantly higher than that which was observed in insects injected with symbiotic bacteria.

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Entomophthora leyteensis Villacarlos & Keller sp. nov. (Entomophthorales: Zygomycetes) infecting Tetraleurodes acaciae (Quaintance) (Insecta, Hemiptera: Aleyrodidae), a recently introduced whitefly on Gliricidia sepium (Jaq.) Walp. (Fabaceae) in the Philippines.

Entomophthora leyteensis Villacarlos & Keller sp. nov., a species of Entomophthorales infecting the whitefly Tetraleurodes acaciae on Gliricidia sepium in the Philippines is described. Disease prevalence monitored weekly for 8 weeks indicated that the fungus could cause 8-31% infection within the whitefly population. Epizootics due to this fungus occurred in Inopacan, Leyte. Sampling live whitefly adults and dissecting them on glass slides for microscopic examination of fungal structures was found to give a better measure of prevalence than actual counts of infected insect cadavers. E. leyteensis is an important mortality factor for T. acaciae. Some speculations on the origin of the fungus are discussed here.

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Peritrophic membranes and protease activity in the midgut of the malaria mosquito, Anopheles stephensi (Liston) (Insecta: Diptera) under normal and experimental conditions.

Formation and solubility of the peritrophic membranes (pm) in the midgut (stomach) of female Anopheles stephensi depended on salt concentration in the gut lumen. High calcium drastically reduced the solubility of the pm in vitro. Thin-layer chromatography revealed the presence of N-acetylgalactosamine and galactose in the pm. In contrast to Aedes aegypti no N-acetylglucosamine was present indicating significant differences between Anopheles and Aedes mosquitoes with respect to pm formation and composition. Experimental distension of the stomach epithelium did not trigger secretion of proteases, but membrane-bound granules were released from the stomach cells, and pm were formed. When females were fed blood 18 hr after an enema with salt solution, when practically no granules were present in the gut cells, protease activity was increased, and blood was digested normally, but no pm were formed. A functional correlation of the membrane-bound granules and pm formation is postulated. alpha-Amanitin prevented normal digestion, protease activity was only insignificantly increased, and no pm formed after blood intake.

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Ovicide-induced serosa degeneration and its impact on embryonic development in Manduca sexta (Insecta: Lepidoptera).

Eggs of Manduca sexta treated with the ovicide Ov. 165049 turn orange, and the embryos later die. The orange pigmentation is at first confined to the serosa, and is accompanied by pathological changes of serosal cells. Lipid vesicles aggregate and spindle-shaped electron-lucent vesicles-normally forming a single layer below the apical cell surface-greatly accumulate. The mitochondria swell considerably, and their matrices become electron-lucent. Subsequently, the serosal cells develop additional features of necrosis. They form many autophagic vacuoles which contain mostly degradating mitochondria, but also segregated rough endoplasmic reticulum (rER) and glycogen granules. The whole cytoplasm vesiculates, and the cells shrink considerably. The nuclei become less irregular in shape, the chromatin disperses rather evenly whereas the nucleoli persist. Neither chromatin condensation nor the production of apoptotic bodies was observed-further evidence, that the serosal cells die by necrosis rather than apoptosis. At some stage of development the damaged serosa ruptures, retracts from the embryo and forms a sphere beneath it. It is only after the rupture of the serosa, that the embryo also turns orange and disintegrates rapidly. This shows impressively the protective function which the serosa plays for the embryo. Our physiological tests indicate, that the orange pigmentation of the serosa induced by the ovicide results from a disturbance of the tryptophan/ommochrome pathway serving the excretion of potentially toxic metabolites of tryptophan-rich proteins. The results demonstrate first that the serosa represents an important target for ovicide pesticides and second that it plays a vital role as an excretory organ during embryogenesis.

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Induction of ACTH- and TNF-alpha-like molecules in the hemocytes of Calliphora vomitoria (Insecta, Diptera).

Three basic cell types are described in the hemolymph of newly enclosed adult Calliphora vomitoria: prohemocytes, plasmatocytes, and granular cells. In addition, anucleate cellular fragments with some inclusions are observed. Cell division is found only in the prohemocytes. Plasmatocytes and granular cells are capable of in vitro bacterial phagocytosis and take part in capsule formation. Adrenocorticotropin hormone (ACTH)-like molecules are found in both cell types. The staining of the plasmatocytes is related to the functional activation of the cells. Indeed, positive staining is only observed during capsule formation, suggesting an induction of ACTH expression. The behaviour of tumor necrosis factor (TNF)-alpha-like molecules is similar. In conclusion, these data provide further support for the close, evolutionary relationship between the immune and the neuroendocrine systems.

Adrenocorticotropic Hormone↗