The impact of body size on Aedes [Stegomyia] aegypti wingbeat frequency: implications for mosquito identification.

Billions of people are at risk due to mosquito-borne diseases. Ideally, the control of mosquito-borne diseases should integrate mosquito control and surveillance to maximize transmission prevention while minimizing environmental impacts. Mosquito surveillance is often limited in scope by logistical constraints, especially the labour and expertise in identifying captured mosquitoes. Mosquito sounds, primarily the wingbeat frequencies (WBF), have been extensively studied in the literature, often targeting a straightforward assessment of this technology with species identification in laboratory conditions. Optical sensors for measuring the WBF of free-flying mosquitoes are the most recent proposal to automate species identification. However, many of the factors that may influence WBF within and between species have not been fully examined, resulting in failures in the species identification. Here we show that body size and temperature modify the wingbeat frequency of female Aedes [Stegomyia] aegypti Linnaeus (Diptera:Culicidae) and such an optical sensor can capture these alterations. We demonstrate that this study's optical sensor can distinguish wingbeat frequency from large and small mosquitoes at different temperatures. The relationship between WBF and size should be taken into account to improve the accuracy of devices that automatically identify species using WBF.

New thinking for filth fly control: residual, non-chemical wall spray from volcanic glass.

Filth flies are of medical and veterinary importance because of the transfer of disease organisms to animals and humans. The traditional control methods include the use of chemical insecticides. A novel mechanical insecticide made from volcanic glass and originally developed to control mosquitoes (Imergard™ WP; ImG) was investigated for control of adult grey flesh flies, Sarcophaga bullata (Parker), secondary screwworms, Cochliomyia macellaria (F.), and house flies, Musca domestica L. In a modified WHO cone test device, the time to 50% mortality (LT50 ) when applied at 5 g/m2 (tested at 30 °C and 50% relative humidity (rH)) was 7.1, 4.3 and 3.2 h, respectively. When knockdown was included, the LT50 s were 5.5, 1.5 and 2.8 h, respectively. Application rates of 1.25 and greater g/m2 had the shortest LT50 s. The time to the LT50 increased for M. domestica as rH increased, but ImG was still active at the highest rH tested of 70%. Scanning electron micrographs showed ImG was present on all body parts, unlike that for mosquitoes where it was found mostly on the lower legs. These first studies on the use of Imergard WP against flies suggest this could be an alternative method for filth fly control.

Quantifying pteridines in the heads of blow flies (Diptera: Calliphoridae): Application for forensic entomology.

In forensic cases involving entomological evidence, establishing the postcolonization interval (post-CI) is a critical component of the investigation. Traditional methods of estimating the post-CI rely on estimating the age of immature blow flies (Diptera: Calliphoridae) collected from remains. However, in cases of delayed discovery (e.g., when remains are located indoors), these insects may have completed their development and be present in the environment as adults. Adult fly collections are often ignored in cases of advanced decomposition because of a presumed little relevance to the investigation; herein we present information on how these insects can be of value. In this study we applied an age-grading technique to estimate the age of adults of Chrysomya megacephala (Fabricius), Cochliomyia macellaria (Fabricius), and Phormia regina (Meigen), based on the temperature-dependent accumulation of pteridines in the compound eyes, when reared at temperatures ranging from 5 to 35°C. Age could be estimated for all species*sex*rearing temperature combinations (mean r2±SE: 0.90±0.01) for all but P. regina reared at 5.4°C. These models can be used to increase the precision of post-CI estimates for remains found indoors, and the high r2 values of 22 of the 24 regression equations indicates that this is a valid method for estimating the age of adult blow flies at temperatures ≥15°C.

Mosquitoes of field and forest: the scale of habitat segregation in a diverse mosquito assemblage.

Knowledge of the distribution of arthropod vectors across a landscape is important in determining the risk for vector-borne disease. This has been well explored for ticks, but not for mosquitoes, despite their importance in the transmission of a variety of pathogens. This study examined the importance of habitat, habitat edges, and the scale at which mosquito abundance and diversity vary in a rural landscape by trapping along transects from grassland areas into forest patches. Significant patterns of vector diversity and distinct mosquito assemblages across habitats were found. The scale of individual species' responses to habitat edges was often dramatic, with several species rarely straying even 10 m from the edge. The present results suggest blood-seeking mosquito species are faithful to certain habitats, which has consequences for patterns of vector diversity and risk for pathogen transmission. This implies that analysts of risk for pathogen transmission and foci of control, and developers of land management strategies should assess habitat at a finer scale than previously considered.

Development of a universal double-digest RAD sequencing approach for a group of nonmodel, ecologically and economically important insect and fish taxa.

The generation of genome-scale data is critical for a wide range of questions in basic biology using model organisms, but also in questions of applied biology in nonmodel organisms (agriculture, natural resources, conservation and public health biology). Using a genome-scale approach on a diverse group of nonmodel organisms and with the goal of lowering costs of the method, we modified a multiplexed, high-throughput genomic scan technique utilizing two restriction enzymes. We analysed several pairs of restriction enzymes and completed double-digestion RAD sequencing libraries for nine different species and five genera of insects and fish. We found one particular enzyme pair produced consistently higher number of sequence-able fragments across all nine species. Building libraries off this enzyme pair, we found a range of usable SNPs between 4000 and 37 000 SNPS per species and we found a greater number of usable SNPs using reference genomes than de novo pipelines in STACKS. We also found fewer reads in the Read 2 fragments from the paired-end Illumina Hiseq run. Overall, the results of this study provide empirical evidence of the utility of this method for producing consistent data for diverse nonmodel species and suggest specific considerations for sequencing analysis strategies.

Recent Changes in the Local Distribution of Aedes aegypti (Diptera: Culicidae) in South Florida, USA.

Disease transmission is directly tied to the spatial distribution of disease vectors. The distribution of Aedes aegypti (L.) in the United States has diminished since the introduction of Aedes albopictus (Skuse) in the 1980s. However, Ae. aegypti persists in some urban areas, particularly in south Florida. The pattern of habitat segregation of these two species is well documented, but the consistency of this phenomenon over time is unknown. To examine the dynamics of the local distributions of these two species, we studied the spatial pattern of Ae. aegypti and Ae. albopictus over time at a fine landscape scale in Palm Beach County, FL. We compared patterns from 2006-2007 with their distributions in 2013, taking into account abiotic factors of microclimate and land cover. We found evidence for a local shift in Ae. aegypti distribution, but could not attribute this to changes in measured abiotic factors. Alternatively, the interaction between Ae. aegypti and Ae. albopictus that initially resulted Ae. aegypti decline may be being attenuated through natural selection. This study confirms the importance of monitoring the changing ranges of these two important vector species.

Comparison of the vector potential of different mosquito species for the transmission of heartworm, Dirofilaria immitis, in rural and urban areas in and surrounding Stillwater, Oklahoma, U.S.A.

Dirofilaria immitis Leidy (Spirurida: Onchocercidae), or heartworm, is a mosquito-borne nematode that causes a fatal disease in carnivores. Although infection is preventable through prophylactic drugs, compliance and the spectre of resistance suggest vector control is a viable alternative. There were two main objectives in this study: (a) to evaluate the relationships between landscape and social factors and the number and species of heartworm-positive mosquitoes, with a specific focus on the importance of the invasive Asian tiger mosquito, Aedes albopictus (Skuse) (Stegomyia albopicta) (Diptera: Culicidae), and (b) to test the hypothesis that dog heartworm is more prevalent in suburban than in rural areas. To achieve these objectives, mosquitoes were collected from May to November 2010 at 16 rural and 16 urban locations in Payne County, Oklahoma, U.S.A. using three trapping methods that utilized, respectively, resting boxes, carbon dioxide traps and BG Sentinel traps. Urban areas showed greater numbers of Ae. albopictus and a higher overall likelihood of infection with D. immitis. Because many species of mosquito are responsible for heartworm transmission, current prophylactic treatment remains the best method of controlling this parasite.

Implications of saline concentrations for the performance and competitive interactions of the mosquitoes Aedes aegypti (Stegomyia aegypti) and Aedes albopictus (Stegomyia albopictus).

Aedes albopictus (Stegomyia albopictus) (Diptera: Culicidae) has probably supplanted Aedes aegypti (Stegomyia aegypti) throughout most of its historical range in the U.S.A., although Ae. aegypti still exists in large coastal cities in southern Florida. We measured salt concentrations in field containers along an axis perpendicular to the coast and examined intraspecific outcomes in these species under different salt concentrations in a factorial study using varying intra- and interspecific densities in different conditions of salinity to order to determine if salt could mitigate the documented competitive superiority of Ae. albopictus. Salt in field containers declined away from the coast, with maximal values similar to our lower salt concentrations. Egg hatching and short-term survival of pupae and late instars were not affected by salt concentrations; survival of early instars of both species decreased at higher concentrations. In high salt conditions, Ae. aegypti achieved higher survival. In the longterm experiment, both species displayed longer development times. Salt did not affect interactions for either species; Ae. aegypti survived in the highest salt conditions, regardless of density. The tolerance of Ae. aegypti to high salt concentrations may allow it to use coastal containers, although because salt did not mediate interspecific interactions between Ae. aegypti and Ae. albopictus, the ultimate effects of salt on the coexistence of these species or exclusion of either species remain unknown.

Spatial and temporal patterns of abundance of Aedes aegypti L. (Stegomyia aegypti) and Aedes albopictus (Skuse) [Stegomyia albopictus (Skuse)] in southern Florida.

Invasion by mosquito vectors of disease may impact the distribution of resident mosquitoes, resulting in novel patterns of vectors and concomitant risk for disease. One example of such an impact is the invasion by Aedes albopictus (Skuse) [Stegomyia albopictus (Skuse)] (Diptera: Culicidae) of North America and this species' interaction with Aedes aegypti L. (Stegomyia aegypti L). We hypothesized that Ae. aegypti would be found in urban, coastal areas that experience hotter and drier conditions, whereas Ae. albopictus would be more commonly found in suburban and rural areas that are cooler and wetter. In addition, we hypothesized that Ae. aegypti would be more abundant early in the wet season, whereas Ae. albopictus would be more abundant later in the wet season. Urban areas were drier, hotter and contained more Ae. aegypti than suburban or rural areas. Aedes aegypti was relatively more abundant early in the wet season, whereas Ae. albopictus was more abundant in both the late wet season and the dry season. The spatial patterns of inter- and intraspecific encounters between these species were also described. The distribution of these mosquitoes is correlated with abiotic conditions, and with temperature, humidity and the relative availability of rain-filled containers. Understanding the ecological determinants of species distribution can provide insight into the biology of these vectors and important information for their appropriate control.

Effects of combination of leaf resources on competition in container mosquito larvae.

Resource diversity is critical to fitness in many insect species, and may determine the coexistence of competitive species and the function of ecosystems. Plant material provides the nutritional base for numerous aquatic systems, yet the consequences of diversity of plant material have not been studied in aquatic container systems important for the production of mosquitoes. To address how diversity in leaf detritus affects container-inhabiting mosquitoes, we examined how leaf species affect competition between two container inhabiting mosquito larvae, Aedes aegypti and Aedes albopictus, that co-occur in many parts of the world. We tested the hypotheses that leaf species changes the outcome of intra- and interspecific competition between these mosquito species, and that combinations of leaf species affect competition in a manner not predictable based upon the response to each leaf species alone (i.e. the response to leaf combinations is non-additive). We find support for our first hypothesis that leaf species can affect competition, evidence that, in general, leaf combination alters competitive interactions, and no support that leaf combination impacts interspecific competition differently than intraspecific competition. We conclude that combinations of leaves increase mosquito production non-additively such that combinations of leaves act synergistically, in general, and result in higher total yield of adult mosquitoes in most cases, although certain leaf combinations for A. albopictus are antagonistic. We also conclude that leaf diversity does not have a different effect on interspecific competition between A. aegypti and A. albopictus, relative to intraspecific competition for each mosquito.

Differential Survivorship of Invasive Mosquito Species in South Florida Cemeteries: Do Site-Specific Microclimates Explain Patterns of Coexistence and Exclusion?

Within 2 yr of the arrival of the invasive container mosquito Aedes albopictus (Skuse), the previously dominant invasive mosquito Aedes aegypti (L.) disappeared from many Florida cemeteries. At some cemeteries, however, Ae. aegypti populations seem stable despite Ae. albopictus invasion. We sought to understand this variation in the outcome (exclusion, coexistence) of this invasion, given that previous experiments show that Ae. albopictus is the superior larval competitor. We tested experimentally the hypothesis that climate-dependent egg survivorship differs between exclusion and coexistence cemeteries and that differences in invasion outcome are associated with microclimate. Viability of eggs oviposited in the laboratory and suspended in vases at six cemeteries was significantly greater for Ae. aegypti than for Ae. albopictus, and greater in 2001 than in 2006. Cemeteries differed significantly in egg survivorship of Ae. albopictus, but not of Ae. aegypti, which is consistent with the hypothesis that Ae. albopictus suffers site-specific, climate-driven egg mortality that mitigates the competitive superiority of larval Ae. albopictus. Principal component (PC) analysis of microclimate records from vases during the experiments yielded three PCs accounting for >96% of the variance in both years of experiments. Multivariate analysis of variance of the three PCs revealed significant microclimate differences among the six cemeteries and between exclusion versus coexistence cemeteries. Stepwise logistic regression of egg survivorship versus microclimate PCs yielded significant fits for both species, and twice as much variance explained for Ae. albopictus as for Ae. aegypti in both years. Higher mortalities in 2006 were associated with high average daily maximum temperatures in vases, with lethal thresholds for both species at ≈40°C. From 1990 to 2007, vase occupancy by Ae. albopictus increased and that by Ae. aegypti decreased, with increasing seasonal precipitation at one well-sampled cemetery. Results support the hypothesis that locally variable climate-driven mortality of Ae. albopictus eggs contributes to patterns of exclusion of, or coexistence with, Ae. aegypti.

Effects of intraspecific larval competition on adult longevity in the mosquitoes Aedes aegypti and Aedes albopictus.

Larval competition is common in container-breeding mosquitoes. The impact of competition on larval growth has been thoroughly examined and findings that larval competition can lead to density-dependent effects on adult body size have been documented. The effects of larval competition on adult longevity have been less well explored. The effects of intraspecific larval densities on the longevity of adults maintained under relatively harsh environmental conditions were tested in the laboratory by measuring the longevity of adult Aedes aegypti (L.) and Aedes albopictus (Skuse) (Diptera: Culicidae) that had been reared under a range of larval densities and subsequently maintained in high- or low-humidity regimes (85% or 35% relative humidity [RH], respectively) as adults. We found significant negative effects of competition on adult longevity in Ae. aegypti, but not in Ae. albopictus. Multivariate analysis of variance suggested that the negative effect of the larval environment on the longevity of Ae. aegypti adults was most strongly associated with increased development time and decreased wing length as adults. Understanding how larval competition affects adult longevity under a range of environmental conditions is important in establishing the relationship between models of mosquito population regulation and epidemiological models of vector-borne disease transmission.

Epidemiological and ecological characteristics of past dengue virus infection in Santa Clara, Peru.

To determine risk factors associated with dengue (DEN) virus infection among residents of Santa Clara, Peru, a rural Amazonian village near Iquitos, a cross-sectional serological, epidemiological and environmental survey was conducted. Demographic, social and behavioural information was obtained by standardized questionnaire from 1225 Santa Clara residents (61.3%) aged 5 years or older. Additional data were obtained on the environmental variables and immature mosquito species and abundance surrounding each household (n = 248). Sera that had been collected previously by the Peruvian Ministry of Health from residents were tested by an enzyme-linked immunosorbent assay (ELISA) for DEN virus IgG antibody. Antibody identity was verified as DEN by plaque reduction neutralization test. Data on individuals were analysed by univariate and multivariable methods, and independent sample t-tests. Spatial clustering was evaluated by comparing distances among DEN positive households. Overall, antibody prevalence was 29.4 % and more than doubled from the youngest to the oldest age groups, but did not differ by sex. Curiously, length of residence in Santa Clara was negatively associated with DEN virus antibodies. More frequent travel to Iquitos was positively associated with seroprevalence. Residents who obtained water from a river source rather than a local well also had significantly higher antibody prevalence. None of the environmental variables measured at each household corresponded to the patterns of antibody distribution. Of the larval mosquitoes found around residences, all were determined to be species other than Aedes. No evidence of spatial autocorrelation among antibody-positive households was detected. These results strongly suggested that recent DEN virus transmission did not occur in the village and that most infections of residents of this rural village were acquired while visiting the city of Iquitos.