Analysis of trapped mosquito excreta as a noninvasive method to reveal biodiversity and arbovirus circulation

Emerging and endemic mosquito-borne viruses can be difficult to detect and monitor because they often cause asymptomatic infections in human or vertebrate animals or cause nonspecific febrile illness with a short recovery waiting period. Some of these pathogens circulate into complex cryptic cycles involving several animal species as reservoir or amplifying hosts. Detection of cases in vertebrate hosts can be complemented by entomological surveillance, but this method is not adapted to low infection rates in mosquito populations that typically occur in low or nonendemic areas. We identified West Nile virus circulation in Camargue, a wetland area in South of France, using a cost-effective xenomonitoring method based on the molecular detection of virus in excreta from trapped mosquitoes. We also succeeded at identifying the mosquito species community on several sampling sites, together with the vertebrate hosts on which they fed prior to being captured using amplicon-based metabarcoding on mosquito excreta without processing any mosquitoes. Mosquito excreta-based virus surveillance can complement standard surveillance methods because it is cost-effective and does not require personnel with a strong background in entomology. This strategy can also be used to noninvasively explore the ecological network underlying arbovirus circulation.     

Dicer-2-Dependent Generation of Viral DNA from Defective Genomes of RNA Viruses Modulates Antiviral Immunity in Insects

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Survival of West Nile virus‐challenged Southern house mosquitoes,Culex pipiens quinquefasciatus,in relation to environmental temperatures

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.     

Natural vertical transmission of dengue viruses in Aedes aegypti in selected sites in Cebu City,Philippines

We attempted to determine the vertical transmission of dengue virus (DENV) in Aedes aegypti in selected sites in Cebu City, Philippines. Mosquito sub‐adults were collected monthly from households and the field during the wet‐dry‐wet season from November, 2011 to July, 2012 and were laboratory‐reared to adults. Viral RNA extracts in mosquitoes were assayed by hemi‐nested RT‐PCR. Results showed that 62 (36.26%; n=679) out of 171 mosquito pools (n=2,871) were DENV+. The minimum infection rate (MIR) of DENV ranged from 0 in wet months to 48.22/1,000 mosquitoes in April, 2012 (mid‐dry). DENVs were detected in larvae, pupae, and male and female adults, with DENV‐4, DENV‐3, and DENV‐1, in that rank of prevalence. DENV‐1 co‐infected with either DENV‐3 or ?4 or with both in April, 2012; DENV‐3 and ?4 were present in both seasons. More DENV+ mosquitoes were collected from households than in field premises (p<0.001) and in the dry than in the wet season (p<0.05), with significant interaction (p<0.05) between sites and premises but no interaction between sites and seasons (p>0.05). By Generalized Linear Mixed models, the type of premises nested in sites and monthly total rainfall were significant predictors of monthly dengue cases (p<0.05) and not MIR, season, temperature, and relative humidity. Surveillance of DENV prevalence in Ae. aegypti and detecting their natural foci in the dry season provide an early warning signal of dengue outbreak.     

Habitat modification for mosquito control in the Ilparpa Swamp, Northern Territory, Australia

Habitat modification is an established method of effective long-term mosquito management, particularly in salt-marsh environments. It is especially pertinent when mosquitoes are known vectors of life-threatening disease and their larval breeding habitat is in close proximity to residential areas. The Ilparpa Swamp is located less than 10 km from Alice Springs, Northern Territory. Wet season rainfall, often followed by effluent discharges to the swamp from the adjacent sewage treatment plant, create ideal sites for the immature stages of the common banded mosquito Culex annulirostris (Skuse), a major vector of Murray Valley encephalitis (MVEV) and Kunjin (KUNV) viruses. Subsequent to increases in notifications of MVEV disease cases in 2000 and 2001, a drainage system was established in the Ilparpa Swamp in early 2002. This paper evaluates the drainage intervention effects. Results indicate a significant reduction in mosquito numbers following habitat modification, which remain low. There have been no seroconversions in sentinel chickens to MVEV or KUNV and no human infections from these viruses in the Alice Springs urban region since the drains were completed. Habitat modification has successfully reduced mosquito numbers and minimized the risk for mosquito-borne disease to residents in Alice Springs urban and surrounding areas, which has never before been documented in Australia.     

Seasonal pattern of daily activity of Aedes caspius, Aedes detritus, Culex modestus, and Culex pipiens in the Po Delta of northern Italy and significance for vector-borne disease risk assessment

The increasing concern about vector-borne diseases such as West Nile disease in northern Italy motivated our analysis of data on the mosquito fauna and the seasonal and daily flight patterns collected in 1998 in the Po Valley. Collections were performed once a week from May to November, with human landing collections and CO(2) traps. Culex pipiens was present from July to October and showed a clearly unimodal nocturnal flight habit. Culex modestus appeared in July-August and showed a bimodal flight pattern, (main peak during the evening and a minor one in the morning). Aedes caspius was present from May to November (highest densities in July-August) and showed a bimodal flight pattern with a major crepuscular peak and a minor dawn peak in the morning. Aedes detritus was the most abundant species in May, with a crepuscular sharply bimodal flight pattern, particularly according to human landing collections. Sunset and sunrise time, in combination with the solar phase (that determines daylight duration and its trend of changing) were the main factors affecting flight behavior. Temperature, relative humidity, and wind speed differently affected the flight behavior of mosquito females according to the species.     

Hyalomma ticks on northward migrating birds in southern Spain: Implications for the risk of entry of Crimean‐Congo haemorrhagic fever virus to Great Britain

Crimean‐Congo haemorrhagic fever virus (CCHFV) is a zoonotic virus transmitted by Hyalomma ticks, the immature stages of which may be carried by migratory birds. In this study, a total of 12 Hyalomma ticks were recovered from five of 228 migratory birds trapped in Spring, 2012 in southern Spain along the East Atlantic flyway. All collected ticks tested negative for CCHFV. While most birds had zero Hyalomma ticks, two individuals had four and five ticks each and the statistical distribution of Hyalomma tick counts per bird is over‐dispersed compared to the Poisson distribution, demonstrating the need for intensive sampling studies to avoid underestimating the total number of ticks. Rates of tick exchange on migratory birds during their northwards migration will affect the probability that a Hyalomma tick entering Great Britain is positive for CCHFV. Drawing on published data, evidence is presented that the latitude of a European country affects the probability of entry of Hyalomma ticks on wild birds. Further data on Hyalomma infestation rates and tick exchange rates are required along the East Atlantic flyway to further our understanding of the origin of Hyalomma ticks (i.e., Africa or southern Europe) and hence the probability of entry of CCHFV into GB.     

Historical environmental change in Africa drives divergence and admixture of Aedes aegypti mosquitoes: a precursor to successful worldwide colonization?

Increasing globalization has promoted the spread of exotic species, including disease vectors. Understanding the evolutionary processes involved in such colonizations is both of intrinsic biological interest and important to predict and mitigate future disease risks. The Aedes aegypti mosquito is a major vector of dengue, chikungunya and Zika, the worldwide spread of which has been facilitated by Ae. aegypti's adaption to human‐modified environments. Understanding the evolutionary processes involved in this invasion requires characterization of the genetic make‐up of the source population(s). The application of approximate Bayesian computation (ABC) to sequence data from four nuclear and one mitochondrial marker revealed that African populations of Ae. aegypti best fit a demographic model of lineage diversification, historical admixture and recent population structuring. As ancestral Ae. aegypti were dependent on forests, this population history is consistent with the effects of forest fragmentation and expansion driven by Pleistocene climatic change. Alternatively, or additionally, historical human movement across the continent may have facilitated their recent spread and mixing. ABC analysis and haplotype networks support earlier inferences of a single out‐of‐Africa colonization event, while a cline of decreasing genetic diversity indicates that Ae. aegypti moved first from Africa to the Americas and then to Asia. ABC analysis was unable to verify this colonization route, possibly because the genetic signal of admixture obscures the true colonization pathway. By increasing genetic diversity and forming novel allelic combinations, divergence and historical admixture within Africa could have provided the adaptive potential needed for the successful worldwide spread of Ae. aegypti.     

Host‐seeking activity and avian host preferences of mosquitoes associated with West Nile virus transmission in the northeastern U.S.A.

Mosquito host‐seeking activity was studied using a custom‐designed trap to explore: (1) at which time interval of the night adult mosquito abatement would be most effective, and (2) if there exists an avian‐specific host‐seeking preference. Overnight trials using traps baited with dry ice showed that Aedes taeniorhynchus (Wiedemann) was most active at dusk and was then captured throughout the night. In contrast, Culex spp. (Cx. pipiens (Linnaeus) and Cx. restuans (Theobald) delayed most activity until about two h after dusk and were then captured through the night. This pattern suggests that management activities directed at adult Culex spp. would be most effective if initiated well after sunset. Mosquito capture rates in traps baited with birds in net bags were significantly greater than those with empty net bags, indicating that mosquitoes were attracted to the birds and not incidentally being sucked in by the custom trap's strong fan motor (Wilcoxon matched‐pairs signed‐ranks test, n= 24, t= 30, p < 0.05). Regression analysis showed that bird weight influenced mosquito attraction (r²= 0.21, p= 0.02). Trials with paired traps that contained different native bird species showed that Gray Catbirds, Dumatella carolinensis, attracted more mosquitoes than the heavier Northern Cardinals, Cardinalis cardinalis (paired samples t‐test, t= 2.58, df= 7, p= 0.04). However, attractiveness did not differ substantially among bird species, and Gray Catbirds did not attract more mosquitoes than all other birds combined as a group. American Robins, Turdus migratorius (n = 4) were comparable in attractiveness to other bird species, but not enough American Robins were captured for a comprehensive study of mosquito avian preference.     

Anti-mosquito antibodies reduce the susceptibility of Aedes aegypti to arbovirus infection

Aedes aegypti (L.) mosquitoes showed a significant reduction in susceptibility to infection with Ross River virus and Murray Valley encephalitis virus when they were fed on a blood-virus mixture containing rabbit antibodies to mosquito midgut components. Presence of the antibodies did not demonstrably affect virus titres in infected mosquitoes, nor the transmission of virus from infected mosquitoes to vertebrates.