Genetic Variation in North American Black Flies in the Subgenus \u3ci\u3ePsilopelmia\u3c/i\u3e (\u3ci\u3eSimulium\u3c/i\u3e: Diptera: Simuliidae)

Resolution of the genetic heterogeneity of closely related insect species depends on the selection of reliable genetic markers derived from representative specimens. We report the results of a survey of genetic variability in nine species of black flies in the subgenus Psilopelmia Enderlein. Three regions of the mitochondrial genome and an amplicon including the internal transcribed spacer 1 of the nuclear ribosomal RNA gene cluster (ITS1) were amplified using the polymerase chain reaction (PCR), and the amplicons were examined for intraspecific and interspecific polymorphisms. Six of the seven Psilopelmia species that yielded PCR products in the ITS1 PCR reaction were found to generate products that were indistinguishable on the basis of size. Similarly, little interspecific variation was noted in the 16S rRNA amplicon among nine species of Psilopelmia assayed by heteroduplex analysis. In contrast, the remaining regions of the mitochondrial genome exhibited both intra- and inter-specific variation when analyzed by heteroduplex analysis. Information collected from the five amplicons could be employed to develop a classification scheme capable of distinguishing the nine species of Psilopelmia examined

Evaluation of a community-based trapping program to collect simulium ochraceum sensu lato for verification of onchocerciasis elimination

Background: Collection of the black fly vectors of onchocerciasis worldwide relies upon human landing collections. Recent studies have suggested that the Esperanza Window Trap baited with a human scent lure and CO2 had the potential to replace human hosts for the collection of Simulium ochraceum sensu lato in Southern Chiapas focus, Mexico. The feasibility of utilizing these traps in a community-based approach for the collection of S. ochraceum s.l. was evaluated. Methodology/Principal findings: Local residents of a formerly endemic extra-sentinel community for onchocerciasis were trained to carry out collections using the traps. The residents operated the traps over a 60-day period and conducted parallel landing collections, resulting in a total of 28,397 vector black flies collected. None of the flies collected were found to contain parasite DNA when tested by a polymerase chain reaction assay targeting a parasite specific sequence, resulting in a point estimate of infection in the vectors of zero, with an upper bound of the 95% confidence interval 0.13 per 2,000. This meets the accepted criterion for demonstrating an interruption of parasite transmission. Conclusions/Significance: These data demonstrate that Esperanza Window Traps may be effectively operated by minimally trained residents of formerly endemic communities, resulting in the collection of sufficient numbers of flies to verify transmission interruption of onchocerciasis. The traps represent a viable alternative to using humans as hosts for the collection of vector flies as part of the verification of onchocerciasis elimination

Developing GIS-based eastern equine encephalitis vector-host models in Tuskegee, Alabama

<p>Abstract</p> <p>Background</p> <p>A site near Tuskegee, Alabama was examined for vector-host activities of eastern equine encephalomyelitis virus (EEEV). Land cover maps of the study site were created in ArcInfo 9.2^®from QuickBird data encompassing visible and near-infrared (NIR) band information (0.45 to 0.72 μm) acquired July 15, 2008. Georeferenced mosquito and bird sampling sites, and their associated land cover attributes from the study site, were overlaid onto the satellite data. SAS 9.1.4^®was used to explore univariate statistics and to generate regression models using the field and remote-sampled mosquito and bird data. Regression models indicated that <it>Culex erracticus </it>and Northern Cardinals were the most abundant mosquito and bird species, respectively. Spatial linear prediction models were then generated in Geostatistical Analyst Extension of ArcGIS 9.2^®. Additionally, a model of the study site was generated, based on a Digital Elevation Model (DEM), using ArcScene extension of ArcGIS 9.2^®.</p> <p>Results</p> <p>For total mosquito count data, a first-order trend ordinary kriging process was fitted to the semivariogram at a partial sill of 5.041 km, nugget of 6.325 km, lag size of 7.076 km, and range of 31.43 km, using 12 lags. For total adult <it>Cx. erracticus </it>count, a first-order trend ordinary kriging process was fitted to the semivariogram at a partial sill of 5.764 km, nugget of 6.114 km, lag size of 7.472 km, and range of 32.62 km, using 12 lags. For the total bird count data, a first-order trend ordinary kriging process was fitted to the semivariogram at a partial sill of 4.998 km, nugget of 5.413 km, lag size of 7.549 km and range of 35.27 km, using 12 lags. For the Northern Cardinal count data, a first-order trend ordinary kriging process was fitted to the semivariogram at a partial sill of 6.387 km, nugget of 5.935 km, lag size of 8.549 km and a range of 41.38 km, using 12 lags. Results of the DEM analyses indicated a statistically significant inverse linear relationship between total sampled mosquito data and elevation (R²= -.4262; p < .0001), with a standard deviation (SD) of 10.46, and total sampled bird data and elevation (R²= -.5111; p < .0001), with a SD of 22.97. DEM statistics also indicated a significant inverse linear relationship between total sampled <it>Cx. erracticus </it>data and elevation (R²= -.4711; p < .0001), with a SD of 11.16, and the total sampled Northern Cardinal data and elevation (R²= -.5831; p < .0001), SD of 11.42.</p> <p>Conclusion</p> <p>These data demonstrate that GIS/remote sensing models and spatial statistics can capture space-varying functional relationships between field-sampled mosquito and bird parameters for determining risk for EEEV transmission.</p

Identification of human semiochemicals attractive to the major vectors of onchocerciasis

Background: Entomological indicators are considered key metrics to document the interruption of transmission of Onchocerca volvulus, the etiological agent of human onchocerciasis. Human landing collection is the standard employed for collection of the vectors for this parasite. Recent studies reported the development of traps that have the potential for replacing humans for surveillance of O. volvulus in the vector population. However, the key chemical components of human odor that are attractive to vector black flies have not been identified. Methodology/Principal Findings: Human sweat compounds were analyzed using GC-MS analysis and compounds common to three individuals identified. These common compounds, with others previously identified as attractive to other hematophagous arthropods were evaluated for their ability to stimulate and attract the major onchocerciasis vectors in Africa (Simulium damnosum sensu lato) and Latin America (Simulium ochraceum s. l.) using electroantennography and a Y tube binary choice assay. Medium chain length carboxylic acids and aldehydes were neurostimulatory for S. damnosum s.l. while S. ochraceum s.l. was stimulated by short chain aliphatic alcohols and aldehydes. Both species were attracted to ammonium bicarbonate and acetophenone. The compounds were shown to be attractive to the relevant vector species in field studies, when incorporated into a formulation that permitted a continuous release of the compound over time and used in concert with previously developed trap platforms. Conclusions/Significance: The identification of compounds attractive to the major vectors of O. volvulus will permit the development of optimized traps. Such traps may replace the use of human vector collectors for monitoring the effectiveness of onchocerciasis elimination programs and could find use as a contributing component in an integratedvector control/drug program aimed at eliminating river blindness in Africa

Molecular Phylogeny and Typing of Blackflies (Diptera: Simuliidae) That Serve as Vectors of Human or Bovine Onchocerciasis

A subregion of the mitochondrial large subunit (16s) rRNA gene was amplified by polymerase chain reaction (PCR) from nine species of blackflies (Diptera: Simuliidae) which serve as natural or experimental vectors of human or bovine Onchocerca parasites. PCR products from each species of blackfly were tested by directed heteroduplex analysis (DHDA), and their genotypes established according to diagnostic banding patterns of the heteroduplex products. Three alleles of mitochondrial 16s rRNA were found to exist in members of the Simulium (Edwardsellum) damnosum sensu lato complex from West Africa, and two alleles were found in the Neotropical Simulium (Psilopelmia) ochraceum Walker complex and the Simulium (Simulium) metallicum Bellardi complex. Different single alleles were detected in Austrosimulium bancrofti, in English S.(S.)noelleri and in two North American laboratory vectors: Simulium (Psilozia) vittatum Zetterstedt and S.(S.)decorum Walker. Phylogenetic analysis of 16s sequences indicated that blackflies from West Africa and the Americas formed distinct clades. Neotropical onchocerciasis vectors were found to be more closely related to Nearctic and Palaearctic nonvector Simulium species than to the African vectors of onchocerciasis

Genetic Variation in North American Black Flies in the Subgenus \u3ci\u3ePsilopelmia\u3c/i\u3e (\u3ci\u3eSimulium\u3c/i\u3e: Diptera: Simuliidae)

Resolution of the genetic heterogeneity of closely related insect species depends on the selection of reliable genetic markers derived from representative specimens. We report the results of a survey of genetic variability in nine species of black flies in the subgenus Psilopelmia Enderlein. Three regions of the mitochondrial genome and an amplicon including the internal transcribed spacer 1 of the nuclear ribosomal RNA gene cluster (ITS1) were amplified using the polymerase chain reaction (PCR), and the amplicons were examined for intraspecific and interspecific polymorphisms. Six of the seven Psilopelmia species that yielded PCR products in the ITS1 PCR reaction were found to generate products that were indistinguishable on the basis of size. Similarly, little interspecific variation was noted in the 16S rRNA amplicon among nine species of Psilopelmia assayed by heteroduplex analysis. In contrast, the remaining regions of the mitochondrial genome exhibited both intra- and inter-specific variation when analyzed by heteroduplex analysis. Information collected from the five amplicons could be employed to develop a classification scheme capable of distinguishing the nine species of Psilopelmia examined

Salivary gland thrombostasin isoforms differentially regulate blood uptake of horn flies fed on control- and thrombostasin-vaccinated cattle

Thrombostasin (TS) is an anticlotting protein found in saliva of Haematobia irritons (horn flies). The polymorphic nature of the ts gene was first associated with success of horn flies blood feeding on a laboratory host, New Zealand White rabbits. In this study, we report results of similar studies testing blood uptake of horn flies feeding on a natural host, cattle. These studies confirmed the association of ts genotype with blood uptake of horn flies and showed that it was host species specific. In contrast to rabbits, blood uptake volumes of homozygous ts10 horn flies were lower than those of other ts genotypes when fed on control (ovalbumin-vaccinated) cattle. Cattle vaccinated with recombinant protein isoforms, rTS9 or rTB8, resisted horn fly feeding by yielding lower blood volumes compared with flies feeding on control cattle. The specific impact of vaccination, however, varied by ts genotype of flies, Cattle vaccinated with isoform rTS9 resisted flies of ts2, ts9, and tb8 genotype. Vaccination with isoform rTB8 produced resistance to ts8, ts9, and tb8 genotype flies. Horn flies of genotype ts10 were not affected by vaccination with either TS isoform and fed as well on rTS9- and rTB8-vaccinated as on control-vaccinated cattle. These experimental results confirm the efficacy of vaccines targeting horn fly salivary proteins and provide new insight into the dynamics of horn fly-cattle interactions in nature. © 2010 Entomological Society of America