Absence of transovarial transmission of bluetongue virus in Culicoides variipennis: immunogold labelling of bluetongue virus antigen in developing oocytes from Culicoides variipennis (Coquillett)

1. Culicoides variipennis midges were fed on a blood meal containing bluetongue virus (BTV) serotype 11 (BTV-11) and on four subsequent non-infective blood meals at 4-day intervals. 2. Eggs were collected before each blood-feeding and reared to adults. 3. Progeny from each egg batch were incubated for 14 days (20 degrees C, 40-60% RH) before plaque assay. 4. Oocytes from several parent flies were sectioned for immunoelectron microscopy. 5. Thirty-two percent of the parent females tested by plaque assay were positive for BTV. 6. All 993 progeny flies were negative for BTV. 7. BTV antigen was dense in proteid yolk bodies and in the vitelline membrane of the developing oocytes.     

A new Culicoides (Diptera:Ceratopogonidae) of the subgenus Diphaomyia from Peru

A new species of Culicoides of the subgenus Diphaomyia, Culicoides jurbergi Felippe-Bauer, is described and illustrated based on female specimens collected biting man and with light traps in Peruvian Amazonia. The species is compared with its similar congener mirsae Ortiz.     

A new Culicoides from the Amazonian region, Brazil (Diptera: Ceratopogonidae)

A new Neotropical species of biting midge Culicoides (Haematomyidium), C. kampa Felippe-Bauer, Veras & Castellon, is described and illustrated based on female specimens from the Amazonian Region.     

Laboratory Culture and Life History of Heleidomermis magnapapula in Its Host, Culicoides variipennis (Diptera: Ceratopogonidae)

The mermithid parasite Heleidomermis magnapapula was maintained in larvae of the midge Culicoides variipennis for 20 months in enamel pans containing nutrient-rich water and polyester pads as a substrate. Inseminated female mermithids were introduced to the pad surface when the host was in the late second or early third-instar. Host larvae were harvested from the pans 9 days after exposure and held in tap water for nematode emergence. Preparasite yield was positively correlated with female nematode size and averaged 1,267 preparasites/female. Male and female nematodes emerged an average of 12.2 and 13.4 days after host exposure, respectively. Supplemental host food (Panagrellus) during the final days of parasitism did not alter time of emergence. Parasites emerging singly were 64% females, whereas superparasitized hosts yielded males (up to nine/host). Nematode carryover into the adult midge normally occurred at a level of 0.5-2.5%. Parasite load (nematodes/ parasitized individual) in midge adults was lower than that of larvae from the same cohort, and adult midges were more likely to harbor female parasites. Exposure of fourth-instar host larvae resulted in higher levels of adult parasitism (up to 17%).     

Effects of temperature on virogenesis of bluetongue virus serotype 11 in Culicoides variipennis sonorensis

Abstract. Culicoides variipennis sonorensis females were fed bluetongue virus serotype 11 mixed in sheep blood and were held at constant temperatures of 32, 27, 21 and 15^oC. Virogenesis, as measured by enzyme-linked immunosorbent assay (ELISA), proceeded significantly faster at higher temperatures. Based on ELISA absorbance ≥0.2, some flies first were categorized as infected after 1 day, 2 days and 4 days at 32, 27 and 21^oC, respectively. Peak levels of virus antigen were seen after 5–7, 7–13 and 18–22 days for flies held at 32, 27 and 21^oC, respectively. There was no significant virus replication in flies held at 15^oC for 22 days, but latent virus replicated and was detected easily (44% infection) 4–10 days after these flies were transferred to 27^oC. The implications for temperature effects on bluetongue epizootiology are discussed.     

A RNA virus in cells from Culicoides variipennis

A virus was detected in cells (designated CuVa) cultured from one laboratory colony of the biting midge, Culicoides variipennis. By electron microscopy (30 nm), nonenveloped, icosahedral virions arranged separately and in crystalline matrix arrays were seen in the cytoplasm but not in the nucleus of CuVa cells. Separation by 10% polyacrylamide gel electrophoresis revealed multiple bands of viral-induced double-stranded RNA. Inoculation of this virus onto different cell lines and intracranially into suckling mice revealed no detectable pathology. Immunoperoxidase staining using polyclonal antibody determined that the virus is infectious to toad cells, bovine endothelial cells, bovine kidney cells, mosquito cells, and cells (designated KC) initiated from another laboratory colony of C. variipennis. KC cells infected with this virus were coinfected with bluetongue virus with no decrease in bluetongue virus titer.     

Description of the Pupa of Culicoides crucifer Clastrier (Diptera: Ceratopogonidae)

The pupa of Culicoides crucifer Clastrier is described, illustrated and photomicrographed by using binocular microscope and phase-contrast microscopy from material collected in an artificial container in Manaus, Brazil. The pupa shows features typical of pupae occurring in calm and clean waters, and it is compared with its similar congeners of the subgenus Haematomyiidium, Culicoides annuliductus Wirth and Culicoides debilipalpis Lutz.     

Effects of seawater components on immature Culicoides molestus (Skuse) (Diptera: Ceratopogonidae)

Abstract Following analysis of beach sites and an indication that seawater components might influence larval occurrence, we studied the impact of increasing salinity and seawater concentration on survival of fourth-instar larvae of the canal biting midge, Culicoides molestus . While NaCl had little effect on immature survival, increasing the concentration of seawater increased mortality prior to the adult stage. Seawater at three and four times the normal concentration killed all immatures. Artificial elevation of seawater concentration in the sandy substrate preferred by larvae, therefore, has the potential to reduce immature midge survival. Diet also affected survival, with higher mortality of immatures that were fed fish-food flakes compared with those that were fed live nematodes.     

A new Culicoides (Diptera: Ceratopogonidae) of the reticulatus species group from Brazilian Amazon Region

A new species of the Culicoides reticulatus species group, Culicoides kuripako Felippe-Bauer, is described and illustrated based on females and male specimens from the states of Amazonas and Pará, Brazil. The new species is compared with its similar sympatric congener, Culicoides paucienfuscatus Barbosa.     

Five new species of Culicoides (Diptera: Ceratopogonidae) of the Nansei Islands.

Five new species of Culicoides of the Nansei Islands are described: C. sasai sp.n. from Amami-oshima Is., C. toshiokai sp.n. from Ishigaki Is., C. iriomotensis sp.n. from Iriomote Is., C. flavipunctatus sp.n. from Yonaguni Is., and C. yaeyamaensis sp.n. from the latter three islands. C. toshiokai sp.n. and C. iriomotensis sp.n. are tree-hole breeders, such as C. dendrophilus Amosova, which is common in similar habitats extending from Okinawa to Honshu.