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Biological and molecular studies of a cypovirus from the black fly Simulium ubiquitum (Diptera: Simuliidae).

A cypovirus from the black fly Simulium ubiquitum (SuCPV) was isolated and examined using biological and molecular techniques. SuCPV produces small (typically 0.25mum), polyhedral shaped inclusion bodies (polyhedra), in which the virus particles become multiply embedded. SuCPV is the third cypovirus isolated from Diptera, but the first from Simuliidae that has been characterized using molecular analyses. SuCPV has a genome composed of 10 segments of dsRNA, with an electrophoretic migration pattern that is different from those of recent UsCPV-17 and CrCPV-17 isolates from the mosquitoes Uranotaenia sapphirina and Culex restuans, respectively. The SuCPV electropherotype appears to show significant differences from those of the previously characterized lepidopteran cypoviruses. Sequence analysis of SuCPV segment 10 shows that it is unrelated to either of the two CPV isolates from Diptera or to the CPV species for which Seg-10 has been previously characterized from Lepidoptera. A comparison of the terminal regions of SuCPV genome segments to those of CPV-1, 2, 4, 5 14, 15, 16, 17, 18, and 19 also revealed only low levels of conservation. We therefore, propose that SuCPV is classified within a new Cypovirus species, which we have tentatively identified as Cypovirus-20. We have therefore referred to this virus isolate as S. ubiquitum CPV-20 (SuCPV-20).

Amino Acid Sequence↗

Comparative larvicidal activities of the four Bacillus thuringiensis serovars against a chironomid midge, Paratanytarsus grimmii (Diptera: Chironomidae).

The four reference strains of Bacillus thuringiensis, known to be pathogenic for mosquito larvae of the family Culicidae (Diptera), were examined for their larvicidal activities against a chironomid species, Paratanytarsus grimmii (Diptera: Chironomidae). Of the four, the type strain of B. thuringiensis serovar israelensis (H14) exhibited the highest activity against both early- and late-instar larvae, while the type strain of serovar kyushuensis (H11ac) and the strain 73-E-10-2 (H10, serovar darmstadiensis) showed relatively moderate toxicities. The strain HD-1 of serovar kurstaki (H3abc) had no larvicidal activity on this chironomid midge.

Animals↗

Identification of bloodmeals in haematophagous Diptera by cytochrome B heteroduplex analysis.

We developed a DNA assay for bloodmeal identification in haematophagous insects. Specific host cytochrome B gene sequences were amplified by PCR and classified on the basis of their mobility in a heteroduplex assay. In the blackfly Simulium damnosum s.l. (Diptera: Simuliidae), human cytochrome B DNA sequences were identifiable up to 3 days following ingestion of the bloodmeal. In the tsetse Glossina palpalis (Diptera: Glossinidae) collected from tsetse traps in Ivory Coast, bloodmeals were identified as taken from domestic pigs on the basis of their heteroduplex pattern and DNA sequence. Evidently the cytochrome B sequence shows sufficient interspecific variation to distinguish between mammalian host samples, while exhibiting minimal intraspecific variation. The stability of DNA in bloodmeals, for several days post-ingestion by haematophagous insects, allows PCR-HDA assays to be used reliably for host identification.

Animals↗

Horn fly (Diptera: Muscidae) saliva targets thrombin action in hemostasis.

The horn fly, Hematobia irritans (L.), is an important pest of livestock because the adult stage of both sexes are aggressive blood-feeders. Remarkably, even though horn fly adults feed recurrently on their hosts as ectoparasites, these flies lack the ADP-responsive antiplatelet aggregation and vasodilatory antihemostatic systems described for other blood-feeding Diptera. Horn fly salivary gland extracts do interfere with the normal coagulation process as demonstrated by the recalcification time assay. Using this as a baseline, the effects of saliva on recalcification time, activated partial thromboplastin time, prothrombin time, and thrombin time were measured to determine which arm(s) of the coagulation cascade might be impacted. Factor-deficient plasma assays also were used to measure possible perturbations in clotting. Gland-free saliva delayed the recalcification time as well as the activated partial thromboplastin time, prothrombin time, and thrombin time. Saliva also further delayed clotting times of plasmas deficient in factor V, factor VIII, and factor XIII, indicating that other factors in the coagulation cascade were inhibited. Although horn fly saliva did not alter the ability of deficient plasma reconstituted with factor X to clot, it did inhibit deficient plasma reconstituted with factor II (thrombin). Antithrombin activity in saliva was confirmed by its ability to interfere with thrombin hydrolysis of fibrinogen, its normal substrate, and by its inhibition of thrombin action on a chromagenic substrate that mimics the hydrolytic site of fibrinogen. Thus, horn fly saliva contains a factor that specifically targets thrombin, a key component in the coagulation cascade. While the biochemical mechanisms of inhibition may vary, this antihemostatic characteristic is shared with other zoophilic Diptera such as black flies, Simulium spp., and tsetse, Glossina morsitans morsitans Westwood, that feed on ungulates.

Animals↗

Diptera as parasitoids.

Parasitoids in the insect order Diptera include an estimated 16,000 species, or approximately 20% of the total number of species with this life-style. Parasitoids in this order are exceedingly diverse in both their habits and evolutionary origins, which makes them an underutilized but highly suitable group for quantitative studies of character convergence and adaptive radiation. This review focuses on several aspects of the bionomics of dipteran parasitoids that have received little comprehensive treatment, including processes associated with host location and attack, patterns of host use, and the evolutionary and ecological consequences of host-parasitoid interactions. Throughout the review we contrast the patterns found within the parasitic Diptera against those found in the better studied parasitic Hymenoptera. We conclude that more intensive study of dipteran parasitoids is required before we can understand the general conditions that favor the evolution of insect parasitoids and the truly magnifying themes of their behavior and ecology.

Journal Article↗

Relationships of phlebotomine sand flies (Diptera).

The status of phlebotomine sand flies in relationship to the family Psychodidae (Diptera) is reviewed. It is concluded that sand flies should be given familial recognition as Phlebotomidae, divided into the subfamilies Phlebotominae and Bruchomyiinae. A comparison is made between the evolution of Psychodidae and Phlebotomidae, and it is concluded that the two families represent contrasting evolutionary experiments at an early stage of the diversification of Diptera.

Animals↗

Nonhost status of commercial Persea americana 'Hass' to Anastrepha ludens, Anastrepha obliqua, Anastrepha serpentina, and Anastrepha striata (Diptera: Tephritidae) in Mexico.

The objective of this study was to determine the host status in Mexico of commercially cultivated and marketed avocado, Persea americana (Mill.), 'Hass' to Anastrepha ludens (Loew), Anastrepha obliqua (Macquart), Anastrepha serpentina (Wiedemann), and Anastrepha striata (Schiner) (Diptera: Tephritidae). Experiments in Michoacán, Mexico, were carried out in six orchards located at three altitudes above sea level during two times (August-October 2001 and April-June 2002). They included choice ('Hass' avocado plus natural host) and no-choice foraging behavior tests on trees under field cages; no-choice, forced infestation trials on caged, fruit-bearing branches in the field, and with individual fruit under laboratory conditions; infestation trials using 'Hass' avocados left unprotected over 1 and 7 d on the ground of orchards; studies to ascertain depth of oviposition and determine egg hatchability; and experiments to determine susceptibility by using time elapsed since removal of fruit from tree as the experimental variable. We trapped adult Anastrepha (n = 7,936) in all orchards and dissected fruit (n = 7,695) from orchards and packing houses (n = 1,620) in search of eggs or larvae. Most (96.7%) A. ludens, A. obliqua, A. striata, and A. serpentina adults were captured in low-elevation orchards. No eggs or larvae were detected in any of the fruit from foraging behavior studies or dissected fruit from orchards or packing houses. Of 5,200 mature, intact fruit on trees in the field forcibly exposed to no-choice female oviposition activity (five females/fruit), we only found four fruit infested by A. ludens but no adults emerged. 'Hass' avocados only became marginally susceptible to attack by A. ludens (but not A. obliqua, A. serpentina, and A. striata) 24 h after being removed from the tree. Fruit placed on the ground in orchards (n = 3,600) were occasionally infested by Neosilba batesi (Curran) (Diptera: Lonchaeidae), a decomposer, but not Anastrepha spp. Based on our results, commercially cultivated and marketed P. americana 'Hass' should not be considered a natural host of A. ludens, A. obliqua, A. striata, and A. serpentina in Mexico.

Animals↗

Adult Fannia benjamini complex (Diptera: Muscidae) activity in southern California and use of CO2 as an attractant.

Diurnal activity of host-seeking "canyon flies" (Fannia benjamini complex) (Diptera: Muscidae) was determined on a warm, sunny day during their peak seasonal activity period (early July) in the coastal mountain community of La Habra Heights in Los Angeles County, California. High levels of activity persisted for several hours in the morning and evening, but peak abundance was within an hour after sunrise and an hour before sunset, when >600 flies (mainly Fannia conspicua Malloch) could be collected in 5 min from a person using a sweep net. Host-seeking activity was low during midday hours, when flies apparently were seeking shelter from the heat, and activity ceased after sunset. Potential bait materials, including some known to elicit a response by other host-seeking Diptera (water, rabbit feces, egg bait, milk bait, Limburger cheese, ethanol, and CO2) were tested for "canyon fly" response using CDC-type suction traps (without light). CO2 resulted in significantly higher capture of female "canyon flies" (up to approximately 2,000 flies per trap in a 6-h period) relative to traps baited with other materials or with no bait. Host-seeking activity in relation to distance from a putative developmental site was evaluated. The proportional capture of flies in CO2-baited suction traps was significantly explained by distance from a residential area planted with Aptenia cordifolia (L.) (Aizoaceae; red apple), a ground cover plant that is a developmental site for F. conspicua. Proportional trap capture rapidly decreased as distance from the residential area increased. Implications of these studies for "canyon fly" control are discussed.

Animals↗

Autophagy is required for the degeneration of the ovarian follicular epithelium in higher Diptera.

Autophagy is a major pathway for the degradation of long-lived proteins and cytoplasmic organelles and an essential part of programmed cell death, as well. Our findings indicate that programmed cell death of the ovarian follicle cells in the higher Diptera species Bactrocera oleae and Ceratitis capitata manifests features of autophagic cell death. The follicle cells during the developmental stage 14 contain autophagic vacuoles and they do not exhibit caspase activity in any area of the egg chamber. Their nuclei are characterized by condensed chromatin, accompanied with high-but not low-molecular weight DNA fragmentation events exclusively detected in distinct cells of the anterior pole. The above results are likely associated with the abundant phagocytosis observed at the entry of the lateral oviducts, where numerous cell bodies are massively engulfed by epithelial cells. The similarity of the cell death process among B. oleae, C. capitata and Drosophila melanogaster species strongly suggests that autophagy-mediated cell death is conserved in higher Diptera species.

Animals↗

[Landscape and mass-attack of Simuliidae-Diptera on man and domestic animals].

Mass-attack of Simuliidae-Diptera on man and domestic animals should not be considered only as a consequence of quality of running freshwater, but also as an effect of the actual state of the surrounding area. The territory, in fact, impacts the qualitative properties of a water body and can trigger adverse phenomena, especially so when modified by significant anthropic activities. In different locations the native settings have been replaced by deforestation, intensive farming and zootechnological activities. This paper sets forth the results of some investigations carried out further to a mass attack of Simuliidae-Diptera on man at different sites of the national territory. These results have been re-assessed taking into account the areas of interest and the modifications that they have undergone.

Animals↗

[The small halophilic zygopteric odonate, Mortonagrion hirosei, of central Japan, a predator utilizable against tiny stinging diptera of coastal salt marshes, especially Ceratopogonidae of the genera Culicoides and Oecacta, pests of sea shores in southwestern U.S.A. and Caribbean area].

Among stinging diptera pullulating in coastal salt marshes Ceratopogonidae gnats (mainly of genus Culicoides and Oecacta) are especially troublesome, particularly in Southeastern U. S. A. and Caribbean area, escaping attacks of most predators by their tiny size. But the zygopteric odonate insect Mortonagrion hirosei is well fitted for hunting those minute diptera, by its tiny size and its behaviour, seeking shelter between halophilic plants (2 facts explaining that it was not discovered in central Japan before 1971...). Its larvae, living in brackish waters of coastal lagoons, can devour those of Ceratopogonidae and at least young stages of those of Mosquitoes whose some halophilic species are dangerous vectors of diseases. According to similarity of climates M. hirosei can certainly thrive in Southeastern U. S. A., and probably in Southern Europe. At lower latitudes problems for completion of annual cycle could perhaps arise from lack of hivernal cooling. It is necessary to make at the world scale methodical researches for other species of zygopteric odonates of similar ecology which could exist in other countries, both for avoiding harmful competition of introduced M. hirosei with native species still unknown and for fulfilling the same ecological function in areas of climate no suitable for this Japanese insect. Introduction of M. hirosei in new geographic areas would be very useful, too, for protection of this interesting species threatened by human activities in its natural biotopes, made of discontinuous and rather little areas.

Animals↗

Programmed cell death in Bradysia hygida (Diptera, Sciaridae) salivary glands presents apoptotic features.

In this work, we present biochemical and morphological evidence that the final steps of programmed cell death (PCD) in the salivary glands of the inferior Diptera, Bradysia hygida, present apoptotic characteristics. In B. hygida, elimination of salivary glands is preceded by the establishment of a typical pattern of protein synthesis; increase in caspase activity; decrease in cell volume; nuclear pyknosis; nuclear DNA breakage; changes in the actin cytoskeleton; and most importantly, destruction of giant cells via formation of apoptotic bodies containing broken DNA or cytoplasm remains. Thus, elimination of B. hygida salivary glands by this process suggests that such mode of PCD is also involved in the destruction of entire organs in insects and, therefore, adds more complexity to the regulation of tissue elimination during development.

Actins↗

Convergent evolution of sexual shape dimorphism in Diptera.

Several patterns of sexual shape dimorphism, such as male body elongation, eye stalks, or extensions of the exoskeleton, have evolved repeatedly in the true flies (Diptera). Although these dimorphisms may have evolved in response to sexual selection on male body shape, conserved genetic factors may have contributed to this convergent evolution, resulting in stronger phenotypic convergence than might be expected from functional requirements alone. I compared phenotypic variation in body shape in two distantly related species exhibiting sexually dimorphic body elongation: Prochyliza xanthostoma (Piophilidae) and Telostylinus angusticollis (Neriidae). Although sexual selection appears to act differently on male body shape in these species, they exhibited strikingly similar patterns of sexual dimorphism. Likewise, patterns of within-sex shape variation were similar in the two species, particularly in males: relative elongation of the male head capsule, antenna, and legs was associated with reduced head capsule width and wing length, but was nearly independent of variation in thorax length. However, the two species presented contrasting patterns of static allometry: male sexual traits exhibited elevated allometric slopes in T. angusticollis, but not in P. xanthostoma. These results suggest that a shared pattern of covariation among traits may have channeled the evolution of sexually dimorphic body elongation in these species. Nonetheless, static allometries may have been shaped by species-specific selection pressures or genetic architectures.

Animals↗

Structure and morphogenesis of the eggshell and micropylar apparatus in the olive fly, Dacus oleae (Diptera: Tephritidae).

The egg of the olive fly, Dacus oleae (Diptera, Tephritidae), is laid inside olives and the larva eventually destroys the fruit. The oocyte is surrounded by several distinct layers which are produced during choriogenesis. The chorion covering the main body of the egg outside of the vitelline membrane includes a "wax" layer, an innermost chorionic layer, an endochorion consisting of inner and outer layers separated by pillars and cavities similar to their counterparts in Drosophila melanogaster, as well as inner and outer exochorionic layers. The anterior pole is shaped like an inverted cup, which is chiefly hollow around its base and has very large openings communicating with the environment. Holes through the surface of the endochorion result from deposition of endochorionic substance around follicular cell microvilli. An opening at the apex of the cup provides an entrance for sperm entering the micropylar canal, which traverses the endochorion and continues into a "pocket" in a thickened vitelline protrusion. The micropylar canal is formed by deposition of endochorion and vitelline membrane around an elongated pair of follicular cell extensions. These extensions later degenerate and leave an empty canal about 5 microns in diameter and the narrower pocket about 1 micron in diameter. Respiration is thought to be facilitated by openings at the base of the anterior pole as well as by openings through the "plastron" around the main body of the shell.

Animals↗

Toxicity of the entomopathogenic fungus, Metarhizium anisopliae (Deuteromycotina: Hyphomycetes) to adult females of the blowfly Lucilia sericata (Diptera: Calliphoridae).

The fungus Metarhizium anisopliae (Metchnikoff) (Deuteromycotina: Hyphomycetes) is a natural component of soil flora world-wide and is a causal agent of the green muscardine diseases of insects. The use of this pathogen as a potential biocontrol agent against adult females of the blowfly, Lucilia sericata (Meigen) (Diptera: Calliphoridae) was considered. The blowfly L sericata is an economically important agent of cutaneous myiasis of sheep, particularly in northern Europe. Exposure of flies to suspensions of fungal spores by immersion, topical application or tarsal contact with treated surfaces all resulted in high levels of fatal infection. Spores suspended in silicone oil resulted in higher levels of infection (50-70%) than those formulated in a 0.3 glitre(-1) solution of the detergent Tween 80 (10-20%). Spore concentration had a significant effect on levels of infection of flies, with the highest levels of mortality (64%) resulting from a suspension of 1 x 10(7) conidia ml(-1). The duration of tarsal contact had a significant effect on levels of infection. Mean infection levels of 30% were observed following exposure of free-flying adults to a single spore-treated, 5 x 15 cm2 cloth surface suspended from the roof of a cage (30 cm cube). The implications of the results for the potential use of M anisopliae in the biocontrol of blowflies are discussed.

Animals↗

Biochemical and immunocytochemical analysis of vitellogenesis in the olive fruit fly Dacus (bactrocera) oleae (Diptera: Tephritidae).

Synthesis and selective accumulation of the major yolk proteins in the developing oocytes of the species Dacus oleae (Diptera: Tephritidae) was studied biochemically and by immunoelectron microscopy. In the hemolymph of adult females, two yolk proteins precursors (or vitellogenins) have been detected. They each exhibit a similar molecular weight and isoelectric point to their respective mature yolk proteins (or vitellins), while electrophoretic analysis of their synthetic profile shows that their levels in the hemolymph increase rapidly during development. Immunogold electron microscopy of ovarian sections, revealed that the hemolymph vitellogenins reach the oocyte through enlarged inter-follicular spaces and demonstrated vitellogenin synthesis by the follicle cells of the vitellogenic follicles. The newly synthesized vitellogenins follow a distinct secretory pathway into these cells as compared to other components being synthesized at the same time (e.g. the vitelline envelope proteins), since they were found in secretory vesicles that appeared to be differentiated from those destined to participate in the vitelline envelope. The vitellogenin-containing vesicles exocytose their contents directionally into the follicle cell/vitelline envelope boundary, and subsequently the vitellogenins diffuse among the gaps of the forming vitelline envelope and reach the oocyte plasma membrane. Their internalization by the oocyte includes the formation of an endocytic complex consisting of coated pits, coated vesicles, endosomes, transitional yolk bodies, and finally mature yolk bodies, in which the storage of the vitellins and other yolk proteins occur. These results are discussed in relation to data obtained from other Dipteran species.

Animals↗

Histopathological effects of tannic acid on the midgut epithelium of some aquatic Diptera larvae.

The impact of tannins on larval Nematocera was investigated by an extensive survey of the relative toxicity of tannic acid in Diptera larvae representative of mosquito communities from alpine hydrosystems (Culicidae, Chaoboridae, Chironomidae, and Simuliidae) together with a nonindigenous vector competent Culicidae species. Bioassays indicate that exposure to tannic acid at concentrations from 0.25 to 4 mM is deleterious for Culex pipiens, Simulium variegatum, and Chironomus annularius, but not for Aedes, Anopheles, Culiseta, and Chaoborus species. Histopathological observations reveal that, among the target organs of tannic acid, mainly the midgut epithelium is affected by treatment. However, the extent of degeneration varies according to the taxon, the duration of the treatment, and the concentrations assayed. The vulnerability of epithelial cells differs among cell types, clear cells of the anterior midgut showing symptoms of intoxication before dark cells of the posterior midgut. The toxic effects of tannic acid are discussed, particularly in comparison to those of insecticidal bacteria, in order to evaluate the potential for use of tannins in the regulation of larval populations of dipteran pests.

Animals↗

Phylogenetic analysis of the genus Cheilosia (Diptera, Syrphidae) using mitochondrial COI sequence data.

The genus Cheilosia is one of the most diverse and speciose genera of Syrphidae (Diptera). The phylogenetic relationships of the hoverfly genus Cheilosia was investigated for the first time using molecular data. The mitochondrial protein-coding gene cytochrome c oxidase subunit I (COI) was chosen for sequencing; 1341 characters were obtained for 24 ingroup taxa and these were analyzed with parsimony. The monophyly of the genus Cheilosia was well supported. Current taxonomic division of Cheilosia into two subgenera (sg. Nigrocheilosia and sg. Neocheilosia) and most nonformalized species groups based on morphology were supported by the monophyletic groups identified in the molecular analysis. The phylogenetic informativeness of COI in resolving the subtribal relationships within the tribe Cheilosiini remains ambiguous.

Animals↗