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Schoenoplectus maritimus (alkali bulrush) has desirable attributes, such as a short growth habit (height of mature stands < 1.5 m) and annual senescence, for a potential alternative to tall (height > 3 m) emergent macrophytes in shallow constructed treatment wetlands treating ammonium-dominated wastewater. The effects of different ammonium nitrogen (NH4-N) levels on alkali bulrush growth and its ability to take up nutrients from the wastewater, as well as on mosquito production, across the range of NH4-N found in constructed wetlands of southern California are unknown. We evaluated the effects of enrichment with NH4-N on mosquito production and on the nutrient uptake and growth of alkali bulrush in two studies. Overall, significantly greater numbers (> 50%) of immature mosquitoes (mainly Culex tarsalis) were found in mesocosms enriched with NH4-N than in mesocosms receiving ambient (<0.3 mg/liter) NH4-N. High NH4-N enrichment (up to 60 mg/liter) did not adversely impact the height and stem density of S. maritimus, although a significant decrease in biomass was observed at the highest enrichment level. Nitrogen uptake by alkali bulrush increased directly with NH4-N enrichment, whereas carbon was conserved in the above-ground biomass across the enrichment gradient. Alkali bulrush is recommended for use as part of integrated mosquito management programs for moderately enriched, multipurpose, constructed treatment wetlands that improve water quality as well as provide wetland habitat for waterfowl.
We studied the morphology and morphometry of scutella from six species of the hemipteran genus Meccus to identify new tools to help solve taxonomic problems in closely related insect species of epidemiological relevance. Scutellum samples were observed by scanning electron microscopy and were subjected to morphometric analysis. The results mainly show differences in central depression shape, posterior process, and vestiture. We found significant dimensional differences in scutellum morphometry and a clear sexual dimorphism among species. A combination of morphology and morphometry can be used to differentiate among species of the genus Meccus.
The amplification of mosquito-borne pathogens is driven by patterns of host use by vectors. While each mosquito species is innately adapted to feed upon a particular group of hosts, this “preference” is difficult to assess in field-based studies, because factors such as host defenses and spatial and temporal overlap of mosquitoes and hosts affect which host animals actually get bitten. Here we examined patterns of host use by mosquitoes feeding on caged raptors at a rehabilitation and education center for birds of prey in Alabama, U.S.A. PCR-based techniques were used to determine the host species fed upon. Of 19 raptor species at the facility, seven were found to be fed upon by mosquitoes. Feeding indices and linear regression indicated that no species or family of raptor were significantly preferred over another (R2=0.46). Relative abundance adjusted for bird size explained a statistically significant amount of the variation in relative host use (R2=0.71), suggesting that bird size is an important component of host selection by mosquitoes. These findings support the hypothesis that traits of host animals drive patterns of host use by mosquitoes in nature, an interaction that leads to amplification of mosquito-borne viruses.
The physiological susceptibility to insecticides and the behavioral responses of four wild-caught populations of female Anopheles epiroticus to synthetic pyrethroids (deltamethrin, permethrin, and alpha-Cypermethrin) were assessed. Test populations were collected from different localities along the eastern coast, Trat (TR), Songkhla (SK), and Surat Thani (ST) and one population from the western coast, Phang Nga (PN). Results showed that all four populations of An. epiroticus were susceptible to all three synthetic pyrethroids tested. Behavioral responses to test compounds were characterized for all four populations using an excito-repellency test system. TR displayed the strongest contact excitation (‘irritancy’) escape response (76.8% exposed to deltamethrin, 74.1% permethrin, and 78.4% alpha-Cypermethrin), followed by the PN population (24.4% deltamethrin, 35% permethrin, and 34.4% for alpha-Cypermethrin) by rapidly escaping test chambers after direct contact with surfaces treated with each active ingredient compared with match-paired untreated controls. Moderate non-contact repellency responses to all three compounds were observed in the TR population but were comparatively weaker than paired contact tests. Few mosquitoes from the SK and ST populations escaped from test chambers, regardless of insecticide tested or type of trial. We conclude that contact excitation was a major behavioral response in two populations of An. epiroticus, whereas two other populations showed virtually no escape response following exposure to the three pyrethroids. The explanation for these large unexpected differences in avoidance responses between pyrethroid-susceptible populations of the same species is unclear and warrants further investigation.
The utilization of detritus sources by mosquito larvae during development may significantly affect adult life history traits and mosquito population growth. Many studies have shown invertebrate carcasses to be an important detritus source in larval habitats, but little is known regarding how invertebrate carcasses are utilized by mosquito larvae. We conducted two studies to investigate the rate of detritus consumption and its effect on larval development and life history traits. Overall, we found that Aedes aegypti and Aedes albopictus larvae rapidly consumed larval detritus, while pupal detritus was consumed at a significantly slower rate. We also found that the consumption of larval detritus significantly increased larval survivorship and decreased male development time but did not significantly influence female development time or pupal cephalothorax length for either sex. Our results suggest that the direct consumption of larval detritus can support the production of adults in larval habitats that lack allochthonous detritus inputs or where such organic inputs are insufficient. These studies indicate that different forms of invertebrate detritus are utilized in distinct ways by mosquito larvae, and therefore different forms of invertebrate detritus may have distinct effects on larval development and adult life history traits.
Understanding the factors influencing mosquito distribution is important for effective surveillance and control of nuisance and disease vector mosquitoes. The goal of this study was to determine how trap height and distance to the city center influenced the abundance and species of mosquitoes collected in Adelaide, South Australia. Mosquito communities were sampled at two heights (<2m and ∼10m) along an urban-rural gradient. A total of 5,133 mosquitoes was identified over 176 trap nights. Aedes notoscriptus, Ae. vigilax, and Culex molestus were all more abundant in lower traps while Cx. quinquefasciatus (an ornithophilic species) was found to be more abundant in high traps. Distance to city center correlated strongly with the abundance of Ae. vigilax, Ae. camptorhynchus, Cx. globocoxitus, and Cx. molestus, all of which were most common at the sites farthest from the city and closest to the saltmarsh. Overall, the important disease vectors in South Australia (Ae. vigilax, Ae. camptorhynchus, Ae. notoscriptus, and Cx. annulirostris) were more abundant in low traps farthest from the city and closest to the saltmarsh. The current mosquito surveillance practice of setting traps within two meters of the ground is effective for sampling populations of the important disease vector species in South Australia.
Questing ticks were sampled monthly over a period of 11 months from February, 2011 to December, 2011 at 13 sites in southern Germany using the flagging method. The ticks were identified to species, gender, and stadium. Although both I. ricinus and D. reticulatus were sampled, this study concentrated on I. ricinus, since it was the most abundant tick to be found. Additional weather data (air and soil temperature, relative air humidity, precipitation, sunshine duration) were recorded on each sampling site and the local vegetation described. A total of 14, 394 ticks was collected (7,862 larvae, 5,568 nymphs, 964 adults) and their activity was recorded in order to determine the seasonal activity pattern over different periods of the year. In contrast to the widely accepted pattern of a bimodal seasonal activity in moderate areas with a dominant peak in spring and a minor peak in autumn, a unimodal activity pattern was found for all development stages on six of the 12 sampling sites. Tick abundance was compared to weather variables. Tick host-seeking activity was found to be significantly dependent on the temperature at ground level, precipitation, and sunshine duration as well as relative air humidity. Adult ticks showed a positive correlation with the duration of sunshine, whereas nymphs were mostly unaffected by this phenomenon.
Triatomines (Hemiptera, Reduviidae) are bloodsucking insects involved in the transmission of Trypanosoma cruzi, the causative agent of Chagas disease, an important public health problem in Latin America. The triatomine species found in sylvatic habitats generally play a limited epidemiological role compared to domestic species, but they may act as a reinfestation source of dwellings after insecticide spraying and have to be carefully considered in control strategies of Chagas disease transmission. The objectives of this work were to carry out a survey of the sylvatic triatomine species colonizing Furnariidae nests in a typical area of the Chaco region of Argentina during the winter and to study the parasites and natural enemies associated with the collected triatomines. Sixty-three triatomine specimens were collected from Furnariidae nests (Coryphistera alaudina and Phacellodomus sibilatrix) randomly selected within the study area. Fifty-four were identified as Psammolestes coreodes, seven as Triatoma platensis, and two as Triatoma infestans. Specimens of T. infestans and T. platensis were found in one nest. The first finding of instar nymphs of T. infestans x T. platensis in a sylvatic habitat is reported. For the first time, sylvatic collected specimens of T. platensis were found infected by T. cruzi. Triatoma virus was found in one Ps. coreodes specimen.
The phlebotomine sand flies (Diptera: Psychodidae, Phlebotominae) are vectors of several infectious pathogens. The presence of a sand fly vector is considered to be a risk factor for the emergence of leishmaniasis in temperate Europe. Hence, the occurrence of phlebotomine sand flies and any changes in their distribution is important in determining the potential change in distribution of leishmaniasis in Europe. Therefore, published evidence for a changing distribution of the important phlebotomine sand fly vectors of leishmaniasis and phlebovirus infection in Europe is reviewed. This paper presents evidence of an increasing risk of establishment by sand fly species, especially for the Atlantic Coast and inland parts of Germany, Switzerland, and Austria. In addition to detection in potentially appropriate areas, the findings show areas of potential future establishment of the species. The most important and urgent necessity within the community of entomologists working on phlebotomines is the need to record the extremes of distribution of each species and obtain data on their regional presence/absence along with increased sharing of the data throughout European projects.
The distribution of the West Nile virus (WNV) in the organs and tissues of the mosquito Culex pipiens pallens, a potential vector of WNV in China, was investigated up to 14 days after oral infection. The WNV antigen was detected in paraffin-embedded mosquitoes using immunocytochemistry and viral titers of post-infected mosquitoes determined by plaque assay. Viral titers sharply decreased 24 h post-infection, were undetectable for the first few days, then rose over the course of infection. The first midgut infection appeared after one day, and the overall infection rate (based on midgut infection) was 43.9%. Other tissues, including hindgut, foregut, ovarian follicles, Malpighian tubules, and ommatidia, showed weak WNV antigens as early as three days post-infection. Staining in the salivary glands first appeared after seven days, and the salivary gland infection rate on the 14th day was 37.5%. Specimens with no detectable WNV antigens in any tissues, and with positive results confined to the midgut, anterior midgut, and hindgut, were observed on the 14th day. The route of viral dissemination from the midgut, and the relative importance of amplifying tissues in mosquitoes' susceptibility to infection, were evaluated. The results indicate that Cx. p. pallens has the ability to harbor WNV throughout its alimentary system and that midgut epithelial cells may be the initial site of the replication of this virus in this species.
Gregarines that parasitise phlebotomine sand flies belong to the genus Psychodiella and, even though they are highly host-specific, only five species have been described to date. Their most outstanding features include the unique localisation of the oocysts in the accessory glands of the female host, which ensures contamination of the egg surface during oviposition, and the fact that they naturally parasitise the vectors of Leishmania, causal agent of leishmaniasis. The type species, Ps. chagasi, was first described in Lutzomyia longipalpis, vector of visceral leishmaniasis (VL), from Brazil. We recently reported Ps. chagasi sequences in Lu. longipalpis from Posadas (Misiones, Argentina), an endemic VL location where this gregarine had not been previously recorded. In order to analyse the incidence of Ps. chagasi infections in Lu. longipalpis from this location, the aim of this study was to develop a diagnostic assay for sand fly gregarine parasites in Lu. longipalpis. For this, we designed primers using the Ps. chagasi sequences we previously identified and performed an in vitro validation by PCR amplification of the original sand fly samples. Their specificity and sensitivity as diagnostic primers were subsequently confirmed by PCR reactions using total DNA extracted from naturally infected Lu. longipalpis from the same location (Posadas, Argentina).
A 3-chamber in-line olfactometer designed for use with sand flies is described and tested as a high-throughput method to screen honeys for attractiveness to Phlebotomus papatasi (four geographic isolates), P. duboscqi (two geographic isolates), and Lutzomyia longipalpis maintained in colonies at the Walter Reed Army Institute of Research. A diversity of unifloral honey odors were evaluated as a proxy for the natural floral odors that sand flies may use in orientation to floral sugar sources in the field. In the 3-chamber in-line olfactometer, the choice modules come directly off both sides of the release area instead of angling away as in the Y-tube olfactometer. Of the 25 honeys tested, five had a significant attraction for one or more of the sand fly isolates tested. This olfactometer and high-throughput method has utility for evaluating a diversity of natural materials with unknown complex odor blends that can then be down-selected for further evaluation in wind tunnels and/or field scenarios.
Investigations on natural antagonists of mosquito larvae found that micro-crustaceans (e.g., Cladocera) control mosquito populations under experimental conditions. However, their relevance for mosquito control under field situations remains widely unclear because important information about habitat preferences and time of occurrence of crustaceans and mosquito larvae are still missing. In order to fill this knowledge gap, a field study was undertaken in different wetland areas of Saxony, Germany, in different habitats (i.e., grassland, forest, and reed-covered wetlands). We found negative interactions between larvae of Ae. vexans and predatory Cyclopoida (Crustacean: Copepoda), which both were dominant during the first two weeks of hydroperiod, at ponds located at grassland habitats. Larvae of Cx. pipiens were spatially associated with competing Cladocera, but they colonized ponds more rapidly. Populations of Cladocera established from the third week of hydroperiod and prevented Cx. pipiens colonization thereafter. Ostracoda were highly abundant during the whole hydroperiod, but their presence was restricted to habitats of reed-covered wetland at one geographical area. Mosquito larvae hardly occurred at those ponds. In general, we found that ponds at the reed-covered wetlands provided better conditions for the initial development of crustaceans and hence, mosquito larval colonization was strongly inhibited. Grassland habitat, in contrast, favored early development of mosquito larvae. This study showed that micro-crustaceans are relevant for mosquito management but their impact on mosquito larvae varies between species and depends on environmental conditions.
An investigation of the behavioral responses of Aedes aegypti (= Stegomyia aegypti) to various concentrations of essential oils (2.5, 5, and 10%) extracted from hairy basil (Ocimum americanum Linn), ginger (Zingiber officinale Roscoe), lemongrass (Cymbopogon citratus Stapf), citronella grass (Cymbopogon nardus Rendle), and plai (Zingiber cassumunar Roxb) were performed using an excito-repellency test chamber. Results showed that Ae. aegypti exhibited varying levels of escape response in both the contact and noncontact chambers in response to different essential oils. The magnitude of the behaviors changed in a dose-response fashion depending on the percent volume to volume concentration of oil used. A 2.5% concentration of hairy basil oil produced a significantly greater escape response compared to the other extracts at the same concentration (P< 0.05). Oils of ginger, lemongrass, and citronella produced stronger irritant and repellent responses at the median 5% concentration compared to the lowest and highest concentrations. There was marked suppression of escape for both contact and noncontact tests using 10% concentrations of hairy basil, lemongrass, and citronella, with high knockdown for all three oils after 30 min. Hairy basil and lemongrass had the highest insecticidal activity to Ae. aegypti, with LC50 values of 6.3 and 6.7 percent, respectively. We conclude that the essential oils from native plants tested, and likely many other extracts found in plants, have inherent repellent and irritant qualities that should to be screened and optimized for their behavior-modifying properties against Ae. aegypti and other biting arthropods of public health and pest importance.
We investigated the effect of West Nile virus (WNV) infection on survival in two colonies of Culex pipiens quinquefasciatus Say (Diptera: Culicidae) originating from Vero Beach and Gainesville, FL. Mosquitoes were fed West Nile virus-infected blood and checked daily for survival. Exposure to WNV decreased survival among Cx. p. quinquefasciatus from Gainesville relative to unexposed individuals at 31° C. In contrast, exposure to WNV enhanced survival among Cx. p. quinquefasciatus from Vero Beach relative to unexposed individuals at 27° C. These results may suggest that exposure to WNV and associated infection could increase or decrease components of fitness, dependent on environmental temperature and intraspecific variation in Cx. p. quinquefasciatus. The relationship between lifespan (time of death in days) and WNV titer differed in the colonies at 31° C and 27° C, suggesting that intraspecific species variation in response to temperature impacts interactions with WNV. While further work is needed to determine if similar effects occur under field conditions, this suggests intraspecific variation in vector competence for WNV and adult survival of Cx. p. quinquefasciatus, both aspects of vectorial capacity.
Adelfo Sánchez-Trinidad, Félix Ordoñez-Sánchez, Ma. Teresa Valdes-Perezgasga, Francisco J. Sánchez-Ramos, Thomas J. Zavortink, Antonio J. Cortés-Guzmán, Aldo I. Ortega-Morales
Aedes brelandi Zavortink is reported for the first time outside of the United States, where it has been found in northern and central parts of Mexico. Ae. triseriatus (Say) is reported in northern and central Mexico and Ae. zoosophus Dyar and Knab is recorded in southern Mexico. Collection records for these species in northern, central, and southern Mexico showing the current distribution of the Aedes Triseriatus Group are included.
The relationship between climate data and tick questing activity is crucial for estimation of the spatial and temporal distribution of the risk of ticks and tick-borne diseases. This study establishes correlations between selected meteorological variables provided by the Weather Research and Forecasting model (WRF) and the questing activity of Ixodes ricinus nymphs and adults on a regional scale across Lower Silesia, Poland. Application of Generalized Linear Mixed Models (GLMM), built separately for adults and nymphs, showed that solar radiation, air temperature, and saturation deficit appeared to be the meteorological variables of prime importance, whereas the wind speed was less important. However, the effect of meteorological parameters was different for adults and nymphs. The adults are also more influenced by forest cover and the percentage of forest type if compared to nymphs. The WRF model providing meteorological variables separately for each location and day of tick sampling can be useful in studies of questing activity of ticks on a regional scale.
The aim of this study was to investigate blood meal sources of mosquitoes captured in municipal parks in the city of São Paulo, Brazil, and to identify possible associations between mosquito species and their food preferences. Fourteen species of blood hosts of 510 engorged adult female mosquitoes were identified using PCR assays with a vertebrate-specific primer set based on cytochrome b mitochondrial DNA of the following vertebrates: birds, dogs, cats, rodents, humans, and other primates. Mosquitoes were captured using a manual aspirator, CDC traps in the canopy, CDC traps at ground level, and Shannon traps. With the exception of cats, all other vertebrates were used as hosts by mosquitoes in the parks. Statistical analysis failed to show any trend toward association between most culicid species captured and the sources of blood meals. Instead, they revealed random patterns, indicating that the mosquitoes fed on the most abundant or convenient blood meal sources. Although feeding preferences were observed in two species (birds in the case of Cx. nigripalpus and dogs in the case of Cx. quinquefasciatus), our results highlight the opportunistic feeding habits of the female mosquitoes in this study.
We investigated spatial autocorrelation of female Aedes aegypti L. mosquito abundance from BG-Sentinel trap and sticky ovitrap collections in Cairns, north Queensland, Australia. BG-Sentinel trap collections in 2010 show a significant spatial autocorrelation across the study site and over a smaller spatial extent, while sticky ovitrap collections only indicate a non-significant, weak spatial autocorrelation. The BG-Sentinel trap collections were suitable for spatial interpolation using ordinary kriging and cokriging techniques. The uses of Premise Condition Index and potential breeding container data have helped improve our prediction of vector abundance. Semiovariograms and prediction maps indicate that the spatial autocorrelation of mosquito abundance determined by BG-Sentinel traps extends farther compared to sticky ovitrap collections. Based on our data, fewer BG-Sentinel traps are required to represent vector abundance at a series of houses compared to sticky ovitraps. A lack of spatial structure was observed following vector control treatment in the area. This finding has implications for the design and costs of dengue vector surveillance programs.
Numbers of flea (Siphonaptera) species (flea species richness) on individual mammals should be higher on large mammals, mammals with dense populations, and mammals with large geographic ranges, if mammals are islands for fleas. I tested the first two predictions with regressions of H. J. Egoscue's trapping data on flea species richness collected from individual mammals against mammal size and population density from the literature. Mammal size and population density did not correlate with flea species richness. Mammal geographic range did, in earlier studies. The intermediate-sized (31 g), moderately dense (0.004 individuals/m2) Peromyscus truei (Shufeldt) had the highest richness with eight flea species on one individual. Overall, island biogeography theory does not describe the distribution of flea species on mammals in the Great Basin Desert, based on H. J. Egoscue's collections. Alternatively, epidemiological or metapopulation theories may explain flea species richness.
Desmond H. Foley, Yvonne-Marie Linton, J. Freddy Ruiz-Lopez, Jan E. Conn, Maria Anice M. Sallum, Marinete M. Póvoa, Eduardo S. Bergo, Tatiane M. P. Oliveira, Izis Sucupira, Richard C. Wilkerson
The Anopheles albitarsis group of mosquitoes comprises eight recognized species and one mitochondrial lineage. Our knowledge of malaria vectorial importance and the distribution and evolution of these taxa is incomplete. We constructed ecological niche models (ENMs) for these taxa and used hypothesized phylogenetic relationships and ENMs to investigate environmental and ecological divergence associated with speciation events. Two major clades were identified, one north (Clade 1) and one south (Clade 2) of the Amazon River that likely is or was a barrier to mosquito movement. Clade 1 species occur more often in higher average temperature locations than Clade 2 species, and taxon splits within Clade 1 corresponded with a greater divergence of variables related to precipitation than was the case within Clade 2. Comparison of the ecological profiles of sympatric species and sister species support the idea that phylogenetic proximity is related to ecological similarity. Anopheles albitarsis I, An. janconnae, and An. marajoara ENMs had the highest percentage of their predicted suitable habitat overlapping distribution models of Plasmodium falciparum and P. vivax, and warrant additional studies of the transmission potential of these species. Phylogenetic proximity may be related to malaria vectorial importance within the Albitarsis Group.
Field-collected rodents and fleas from ten provinces covering four regions of Thailand were investigated for possible rickettsial pathogen infections. The 257 trapped-rodents belonged to 12 species. Five species of Genus Rattus accounted for 93% of the total capture, of which Rattus exulans and Rattus norvegicus were the two major species caught. All flea specimens, removed from trapped rodents, were identified as Xenopsylla cheopis. The PCR technique was performed on ectoparasite specimens to detect the presence of murine typhus pathogen (Rickettsia typhi) and scrub typhus pathogen (Orientia tsutsugamushi). Thirteen flea specimens (2.6 %) were found to be positive for R. typhi but none for O. tsutsugamushi. An ELISA technique was used to detect the rodent's antibodies against R. typhi and O. tsutsugamushi. Sixty-one rodent serum samples (23.7%) were positive for R. typhi specific IgM, IgG, or both, while 47 of the samples (18.3%) were positive for O. tsutsugamushi. Twenty serum samples from R. norvegicus (7.8%) had detectable antibodies against both R. typhi and O. tsutsugamushi. Our findings revealed the existence of the dual infection of rickettsial pathogens in the same natural hosts.
We investigated the oviposition behavior of Ae. aegypti and Ae. albopictus. In particular we examined whether small-scale site characteristics and the presence of conspecifics or congeners altered oviposition by these mosquitoes. Various combinations of females of the two species were allowed to oviposit inside cages among either vegetation (potted plants) or structural components (wood and concrete blocks). Numbers of eggs deposited per female were compared between species, sides, and treatments. Most significant differences between treatments and species involved differences between single species and mixed species treatments. Ae. aegypti deposited more eggs/female in the vegetation side than in the structure side whereas the opposite pattern was evident for Ae. albopictus. Ae. aegypti females had higher frequency of skip oviposition than Ae. albopictus. An average of 63% of the containers in the two-species treatments contained eggs of both species, with more frequent joint occurrences observed in the treatment with three females of each species than in the treatments with one of each. Our results point to the existence of various interactions between gravid Ae. aegypti and Ae. albopictus females at or near the oviposition sites but further experimental work is necessary to fully characterize the interactions and their specific mechanisms.
A series of laboratory experiments compared predation rates of three native eastern Australian mangrove fish species (Psuedomugil signifer, Hyseleotris galii, Pseudogobius sp.) and the exotic Gambusia holbrooki on 2nd and 4th instar Aedes vigilax larvae, in order to determine their potential as mosquito control agents in mangrove forests. All four species preyed on significant numbers of both 2nd and 4th instar larvae. All showed a similar pattern of larval consumption, gorging on larvae in the first hour of each experiment, before reducing to a relatively constant background feeding rate. Gambusia holbrooki showed the highest larval consumption rates, but is unsuitable as a mosquito control agent due to it being an exotic pest species in Australia. Of the three native species, P. signifer showed the greatest potential as a mosquito control agent, having consumption rates comparable to G. holbrooki, and was the only species that did not show a significant reduction in larval consumption in the night experiments.
The reliability of the length of wing radial vein r2/3 as a character for the morphological discrimination of the two potential arbovirus vectors Culex pipiens s.s. and Cx. torrentium from Germany was reassessed, after this character had been neglected for more than 40 years. Additionally, multivariate morphometric analyses were applied to evaluate wing shape variation between both species. Although high-throughput molecular tools are now available to differentiate the two species, a simple, low-cost routine alternative may be useful in the absence of a molecular laboratory, such as under semi-field conditions. A thin-plate splines transformation confirmed that primarily the shrinkage of vein r2/3 is responsible for the wing differences between the two species. In the bivariate analysis, the r2/3/r3 indices of Cx. pipiens s.s. and Cx. torrentium were 0.185 and 0.289, respectively, resulting in a correct classification of more than 91% of all tested specimens. Using the absolute length of vein r2/3 alone still allowed for more than 90% accurate discrimination. Furthermore, classification accuracy of linear discriminant analysis exceeded 97%.
Four isolates of the entomopathogenic fungus Metarhizium anisopliae were tested for their potential to control the biting midge Culicoides brevitarsis, the principal vector of bluetongue virus in Australia. Adult C. brevitarsis died three to eight days after walking on paper substrate treated with 0.7 g/m2 conidia of any of the isolates, indicating that M. anisopliae has potential as a surface treatment or topical application control strategy. Incorporation of the fungus into freshly excreted cattle dung at rates of between 0.25 and 1 g conidia/kg reduced the emergence of adult midges by up to 98.5% compared to untreated dung indicating that M. anisopliae has the potential to control C. brevitarsis larvae in cattle dung. Three of the isolates produced similar mortality rates on adult and immature C. brevitarsis while the fourth isolate produced lower, but still significant, mortality rates on adult and immature stages.
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