Risk Of Arbovirus Transmission Research Articles

Candidate biomarkers for mosquito age-grading identified by label-free quantitative analysis of protein expression in Aedes albopictus females

We applied a “shotgun” approach based on nanoliquid chromatography–high resolution mass spectrometry associated to label free quantification (LFQ) to identify proteins varying with age, independently from the physiological state, in Aedes albopictus, a mosquito species which in the last decades invaded temperate regions in North America and Europe, creating concerns for associated high nuisance and risk of arbovirus transmission. The combined “shotgun” and LFQ approach was shown to be highly suitable to simultaneously compare several biological samples, as needed in a study aimed to analyze different age-groups and physiological states of adult mosquito females. The results obtained represent the first wide-scale analysis of protein expression in Ae. albopictus females: >1000 and 665 proteins were identified from few micrograms of crude protein extracts of mosquito heads and thoraxes, respectively. Six of these proteins were shown to significantly vary from 2- to 16-day-old females, independently from their physiological state (i.e. virgin, mated, host-seeking, blood-fed, and gravid). Biological significanceMosquito-borne diseases, such as malaria, dengue and other arboviroses, are a persistent cause of global mortality and morbidity, affecting hundreds of thousands of people. Billions of people living in tropical areas are at risk of being bitten every day by an infective mosquito female and the spread of tropical species such as Aedes albopictus to temperate areas is creating alarm in the northern hemisphere. Mosquito longevity is a critical factor affecting mosquito-borne pathogen transmission cycles and the mosquito capacity to transmit pathogens. However, large scale analyses of the age structure of mosquito field populations is hampered by the lack of optimal age-grading approaches. Our findings open new perspectives for the development of reliable, simple and cheap protein-based assays to age-grade Ae. albopictus females and, most likely, other mosquito species of higher medical relevance, such as the main dengue vector, Aedes aegypti, and the major Afrotropical malaria vectors. These assays would greatly contribute to epidemiological studies aimed at defining the actual vectorial capacity of a given mosquito species. Moreover, they would be very valuable in assessing the effectiveness of mosquito control interventions based on the relative ratio between young and old individuals before and after the intervention.

Autochthonous Chikungunya Transmission and Extreme Climate Events in Southern France.

BackgroundExtreme precipitation events are increasing as a result of ongoing global warming, but controversy surrounds the relationship between flooding and mosquito-borne diseases. A common view among the scientific community and public health officers is that heavy rainfalls have a flushing effect on breeding sites, which negatively affects vector populations, thereby diminishing disease transmission. During 2014 in Montpellier, France, there were at least 11 autochthonous cases of chikungunya caused by the invasive tiger mosquito Aedes albopictus in the vicinity of an imported case. We show that an extreme rainfall event increased and extended the abundance of the disease vector Ae. albopictus, hence the period of autochthonous transmission of chikungunya.Methodology/Principal FindingsWe report results from close monitoring of the adult and egg population of the chikungunya vector Ae. albopictus through weekly sampling over the entire mosquito breeding season, which revealed an unexpected pattern. Statistical analysis of the seasonal dynamics of female abundance in relation to climatic factors showed that these relationships changed after the heavy rainfall event. Before the inundations, accumulated temperatures are the most important variable predicting Ae. albopictus seasonal dynamics. However, after the inundations, accumulated rainfall over the 4 weeks prior to capture predicts the seasonal dynamics of this species and extension of the transmission period.Conclusions/SignificanceOur empirical data suggests that heavy rainfall events did increase the risk of arbovirus transmission in Southern France in 2014 by favouring a rapid rise in abundance of vector mosquitoes. Further studies should now confirm these results in different ecological contexts, so that the impact of global change and extreme climatic events on mosquito population dynamics and the risk of disease transmission can be adequately understood.

New adhesive traps to monitor urban mosquitoes with a case study to assess the efficacy of insecticide control strategies in temperate areas.

BackgroundUrban mosquitoes in temperate regions may represent a high nuisance and are associated with the risk of arbovirus transmission. Common practices to reduce this burden, at least in Italian highly infested urban areas, imply calendar-based larvicide treatments of street catch basins – which represent the main non-removable urban breeding site – and/or insecticide ground spraying. The planning of these interventions, as well as the evaluation of their effectiveness, rarely benefit of adequate monitoring of the mosquito abundance and dynamics. We propose the use of adhesive traps to monitor Aedes albopictus and Culex pipiens adults and to evaluate the efficacy of insecticide-based control strategies.MethodsWe designed two novel types of adhesive traps to collect adult mosquitoes visiting and/or emerging from catch basins. The Mosquito Emerging Trap (MET) was exploited to assess the efficacy of larvicide treatments. The Catch Basin Trap (CBT) was exploited together with the Sticky Trap (ST, commonly used to collect ovipositing/resting females) to monitor adults abundance in the campus of the University of Rome “Sapienza” - where catch basins were treated with Insect Growth Regulators (IGR) bi-monthly and Low-Volume insecticide spraying were carried out before sunset - and in a nearby control area.ResultsResults obtained by MET showed that, although all monitored diflubenzuron-treated catch basins were repeatedly visited by Ae. albopictus and Cx. pipiens, adult emergence was inhibited in most basins. Results obtained by ST and CBT showed a significant lower adult abundance in the treated area than in the untreated one after the second adulticide spraying, which was carried out during the major phase of Ae. albopictus population expansion in Rome. Spatial heterogeneities in the effect of the treatments were also revealed.ConclusionsThe results support the potential of the three adhesive traps tested in passively monitoring urban mosquito adult abundance and seasonal dynamics and in assessing the efficacy of control measures. ST showed higher specificity for Ae. albopictus and CBT for Cx. pipiens. The results also provide a preliminary indication on the effectiveness of common mosquito control strategies carried out against urban mosquito in European urban areas.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-015-0734-4) contains supplementary material, which is available to authorized users.

Emergency mosquito control on a selected area in eastern north Carolina after hurricane irene.

Natural disasters such as hurricanes may contribute to mosquito abundance and, consequently, arbovirus transmission risk. In 2011, flooding from Hurricane Irene in eastern North Carolina (NC) resulted in increased mosquito populations that hindered recovery efforts. Budget shortfalls in NC have reduced the functionality of long-term mosquito surveillance and control programs; hence, many counties rely on the Federal Emergency Management Agency for post-disaster mosquito control. This pilot study examines mosquito abundance pre- and post-aerial insecticide spraying at eight study sites in Washington and Tyrrell Counties in rural eastern NC after Hurricane Irene. Percent change was calculated and compared for traps in areas that received aerial pesticide application and those that did not. Traps in spray zones show decreases in mosquito abundance when compared to control traps (treatment: −52.93%; control: 3.55%), although no significant differences (P = 0.286) were found in mosquito abundance between groups. Implications of reactive rather than proactive mosquito control responses are discussed.

Abundance, diversity, and distribution of mosquito vectors in selected ecological regions of Kenya: public health implications

The diversity of mosquito arbovirus vectors was investigated to define regional risk of arbovirus transmission in Kenya. Mosquitoes were sampled between April, 2007 and December, 2010 at thirteen sites across seven administrative provinces and ecological zones. CDC light traps were used to collect mosquitoes while human-landing collection was conducted in five of the sites to target day-feeding Aedes (Stegomyia) species. Over 524,000 mosquitoes were collected and identified into 101 species, 30 of them known vectors of arboviruses endemic to Kenya. Ae. (Neomelaniconion) mcintoshi and Ae. (Aedimorphus) ochraceus were most abundant in Garissa in the arid northeastern province, and Mansonia uniformis and Mn. africana in semi-arid Baringo in the Rift Valley Province. Ae. ochraceus, Mn. africana and Mn. uniformis were also significant in Nyanza Province, while Ae. (Neomelaniconion) circumluteolus predominated in Budalangi, Western Province. Aedes (Stegomyia) aegypti was predominant in Rabai in the Coast Province but insignificant in the western and Nyanza sites. Culex pipiens was abundant in Rift Valley and Nyanza Provinces around the lake shores. This study highlights the potential for emergence and re-emergence of arboviral diseases among vulnerable populations. This calls for comprehensive mapping of vector distribution and abundance for planning focused vector control measures.

The Genome Sequence of Lone Star Virus, a Highly Divergent Bunyavirus Found in the Amblyomma americanum Tick

Viruses in the family Bunyaviridae infect a wide range of plant, insect, and animal hosts. Tick-borne bunyaviruses in the Phlebovirus genus, including Severe Fever with Thrombocytopenia Syndrome virus (SFTSV) in China, Heartland virus (HRTV) in the United States, and Bhanja virus in Eurasia and Africa have been associated with acute febrile illness in humans. Here we sought to characterize the growth characteristics and genome of Lone Star virus (LSV), an unclassified bunyavirus originally isolated from the lone star tick Amblyomma americanum. LSV was able to infect both human (HeLa) and monkey (Vero) cells. Cytopathic effects were seen within 72 h in both cell lines; vacuolization was observed in infected Vero, but not HeLa, cells. Viral culture supernatants were examined by unbiased deep sequencing and analysis using an in-house developed rapid computational pipeline for viral discovery, which definitively identified LSV as a phlebovirus. De novo assembly of the full genome revealed that LSV is highly divergent, sharing <61% overall amino acid identity with any other bunyavirus. Despite this sequence diversity, LSV was found by phylogenetic analysis to be part of a well-supported clade that includes members of the Bhanja group viruses, which are most closely related to SFSTV/HRTV. The genome sequencing of LSV is a critical first step in developing diagnostic tools to determine the risk of arbovirus transmission by A. americanum, a tick of growing importance given its expanding geographic range and competence as a disease vector. This study also underscores the power of deep sequencing analysis in rapidly identifying and sequencing the genomes of viruses of potential clinical and public health significance.

First Attempt To Validate Human IgG Antibody Response to Nterm-34kDa Salivary Peptide as Biomarker for Evaluating Exposure to Aedes aegypti Bites

BackgroundMuch effort is being devoted for developing new indicators to evaluate the human exposure to Aedes mosquito bites and the risk of arbovirus transmission. Human antibody (Ab) responses to mosquito salivary components could represent a promising tool for evaluating the human-vector contact.Methodology/Principal findingsTo develop a specific biomarker of human exposure to Aedes aegypti bites, we measured IgG Ab response to Ae. aegypti Nterm-34 kDa salivary peptide in exposed children in 7 villages of Southern Benin (West Africa). Results showed that specific IgG response presented high inter-individual heterogeneity between villages. IgG response was associated with rainfall and IgG level increased from dry (low exposure) to rainy (high exposure) seasons. These findings indicate that IgG Ab to Nterm-34 kDa salivary peptide may represent a reliable biomarker to detect variation in human exposure to Ae. aegypti bites.Conclusion/SignificanceThis preliminary study highlights the potential use of Ab response to this salivary peptide for evaluating human exposure to Ae. aegypti. This biomarker could represent a new promising tool for assessing the risk of arbovirus transmission and for evaluating the efficacy of vector control interventions.

Biting Activity and Host Attractancy of Mosquitoes (Diptera: Culicidae) in Manzhouli, China

The biting activity and host attractancy of vector mosquitoes are important in assessing the risk of arbovirus transmission, especially where migratory and nonmigrating bird species congregate, such as in Hulun Lake, Manzhouli. In 2009, the population distribution, species diversity, biting activity, and host attractancy of mosquitoes were investigated in Hulan Lake and its associated prairie area. The adult mosquitoes were captured either by human volunteers using aspirators in mosquito nets, by CO2-baited light traps, or by animal-baited traps. In total, 27,004 mosquitoes, representing three genera and 10 species, were collected from Manzhouli, China, in July 2009, of which Aedes dorsalis (Meigen) were most predominant species, followed by Ae. vexans (Meigen). Biting activity peaks by Ae.flavescens (Muller), Ae. dorsalis, and Culex modestus (Facalbi) on human subjects were investigated. Four mosquito species were captured from different animal sheds (sheep, cattle, and goose). Ae. flavescens was more abundant in the cattle shed than in the other two sheds. The Ae. dorsalis in the sheep shed was much higher than in the other animal sheds. The Ae.flavescens collected via chicken-baited traps were significantly higher than those collected via rabbit-baited and pigeon-baited traps. There were no significant differences in the number of Ae. dorsalis and Ae. vexans collected using the three different animal traps.

A geographical information system-based web model of arbovirus transmission risk in the continental United States of America

A degree-day (DD) model of West Nile virus capable of forecasting real-time transmission risk in the continental United States of America up to one week in advance using a 50-km grid is available online at https://sites. google.com/site/arbovirusmap/. Daily averages of historical risk based on temperatures for 1994-2003 are available at 10km resolution. Transmission risk maps can be downloaded from 2010 to the present. The model can be adapted to work with any arbovirus for which the temperature-related parameters are known, e.g. Rift Valley fever virus. To more effectively assess virus establishment and transmission, the model incorporates "compound risk" maps and forecasts, which includes livestock density as a parameter.

Evaluation of the Human IgG Antibody Response to Aedes albopictus Saliva as a New Specific Biomarker of Exposure to Vector Bites

BackgroundThe spread of Aedes albopictus, a vector for re-emergent arbovirus diseases like chikungunya and dengue, points up the need for better control strategies and new tools to evaluate transmission risk. Human antibody (Ab) responses to mosquito salivary proteins could represent a reliable biomarker for evaluating human-vector contact and the efficacy of control programs.Methodology/Principal FindingsWe used ELISA tests to evaluate specific immunoglobulin G (IgG) responses to salivary gland extracts (SGE) in adults exposed to Aedes albopictus in Reunion Island. The percentage of immune responders (88%) and levels of anti-SGE IgG Abs were high in exposed individuals. At an individual level, our results indicate heterogeneity of the exposure to Aedes albopictus bites. In addition, low-level immune cross-reactivity between Aedes albopictus and Aedes aegypti SGEs was observed, mainly in the highest responders.Conclusion/SignificanceAb responses to saliva could be used as an immuno-epidemiological tool for evaluating exposure to Aedes albopictus bites. Combined with entomological and epidemiological methods, a “salivary” biomarker of exposure to Aedes albopictus could enhance surveillance of its spread and the risk of arbovirus transmission, and could be used as a direct tool for the evaluation of Aedes albopictus control strategies.

Detection of Culicoides brevitarsis Activity in Kyushu

Culicoides brevitarsis transmits important ruminant arboviruses, such as Akabane, Aino and bluetongue viruses. The presence of this species has so far been recognized primarily in Okinawa, the southernmost prefecture of Japan. In entomological surveys in 2008 and 2009, C. brevitarsis was collected at 8 sites throughout Nagasaki, Kumamoto and Kagoshima Prefectures. The collection sites are all located near pastures, where the larvae of C. brevitarsis can grow in cattle dung left in the field. C. brevitarsis was confirmed at the same sites in two consecutive years, suggesting that it overwinters in Kyushu. Given the risk of arbovirus transmission, the ecology of C. brevitarsis in Japan, such as its distribution range, seasonal abundance and larval breeding sites, should be investigated in more detail.

Sources of error in the estimation of mosquito infection rates used to assess risk of arbovirus transmission.

Infection rate is an estimate of the prevalence of arbovirus infection in a mosquito population. It is assumed that when infection rate increases, the risk of arbovirus transmission to humans and animals also increases. We examined some of the factors that can invalidate this assumption. First, we used a model to illustrate how the proportion of mosquitoes capable of virus transmission, or infectious, is not a constant fraction of the number of infected mosquitoes. Thus, infection rate is not always a straightforward indicator of risk. Second, we used a model that simulated the process of mosquito sampling, pooling, and virus testing and found that mosquito infection rates commonly underestimate the prevalence of arbovirus infection in a mosquito population. Infection rate should always be used in conjunction with other surveillance indicators (mosquito population size, age structure, weather) and historical baseline data when assessing the risk of arbovirus transmission.

Aedes(Diptera: Culicidae) Vectors of Arboviruses in Mayotte (Indian Ocean): Distribution Area and Larval Habitats

Biological invasions generally induce profound effects on the structure of resident communities. In Mayotte, where Aedes aegypti and Ae. lilii were already present, the recent introduction of Ae. albopictus raises public health concerns because it may affect the risk of arbovirus transmission. Entomological surveys were carried out in six locations on the island following a transect defined by a gradient from urban to rural habitats during a dry and a wet season. A total of 438 larval habitats containing Aedes spp. immature stages were surveyed. We evaluated the characteristics of larval habitats and analyzed the field distribution of Aedes spp. throughout Mayotte using generalized linear models. Artificial containers used for water storage were significantly more productive for Ae. albopictus immature stages than for Ae. aegypti ones. Most of natural larval habitats collected were colonized by Ae. aegypti, and it was also the most common Aedes species encountered in rural habitats. Conversely, Ae. albopictus greatly predominated in urban and suburban habitats and during the dry season. Ae. lilii was uncommon and occurred preferentially in leaf axillae and dead leaves on the ground. Ae. albopictus has rapidly colonized the inhabited areas of Mayotte. A displacement of Ae. aegypti populations by Ae. albopictus populations in urban areas might be happening. The increasing urbanization seems to greatly favor the presence of the invasive species. Thus, arbovirus surveillance programs should focus, as a priority, on areas where this vector is present because of a higher risk of emergence of an epidemic source of arboviruses.

Dengue and Chikungunya viruses in blood donations: risks to the blood supply?

Dengue and Chikungunya viruses in blood donations: risks to the blood supply?

Geographic Distribution and Breeding Site Preference ofAedes albopictusandAedes aegypti(Diptera: Culicidae) in Cameroon, Central Africa

Presence in Cameroon of the recently introduced Aedes (Stegomyia) albopictus (Skuse) in association with the indigenous Aedes aegypti (L.) raises public heath concerns because it might alter the risk of arbovirus transmission. The breeding site and distribution of the two Stegomyia species are updated and reported following entomological surveys carried out in 22 localities throughout Cameroon, with a total of 1,353 containers with water visited. Ae. aegypti was found in every location sampled, showing higher infestation rates in northern Cameroon. Breeding populations of Ae. albopictus were observed in all 19 southern localities, up to the Adamaoua mountains, but the species was not recorded further north. In the area where both species are present, they were often sampled in the same larval developmental sites, suggesting convergent habitat segregation. The most frequently encountered artificial and natural breeding sites were used tires, discarded tins and plastic containers, abandoned car parts, brick holes, dead leaves on the ground, tree holes, and rock pools. Further monitoring of the demographic as well as geographic expansion of Ae. albopictus in this Afrotropical environment and its relationships with indigenous Ae. aegypti should provide insight into the biology of this highly invasive species and help to implement arboviruses surveillance programs in the area.

Spatial Statistical Analysis of Adult Mosquito (Diptera: Culicidae) Counts: An Example Using Light Trap Data, in Redland Shire, Southeastern Queensland, Australia

Many mosquito control agencies use carbon dioxide-baited traps as surveillance tools for adult vector populations. However, decisions regarding the number and location of trap sites and the frequency of collections are often based on logistical issues, and not on the bionomics or spatial distribution of the target species. Therefore, with the aim of providing practical information for adult mosquito surveillance programs, we used an array of 81 carbon dioxide- and octenol-baited lights traps to obtain weekly samples of adult mosquitoes in Redland Shire in southeastern Queensland, Australia. The spatial patterns of four different mosquito species were examined, and positive spatial autocorrelation in trap counts was evident for Ochlerotatus vigilax (Skuse), Coquillettidia linealis (Skuse), and Culex annulirostris Skuse, but not for the container species Ochlerotatus notoscriptus (Skuse). Of the three species that exhibited spatially correlated trap counts, the autocorrelation was greatest in Oc. vigilax at a lag distance of 0-1.5 km, with Moran's I values of 0.30-0.64. Moran's I indices were also positive and statistically significant (P < 0.05) at lag distances of 1.5-3.0 and 3.0-4.5 km on each of the 15 sampling occasions. However, at 3.0-4.5 km the Moran's I values were low, which indicated only weak spatial autocorrelation in trap counts. Universal kriging was used to estimate the numbers of each species at unsampled locations throughout the study area, and leave-one-out cross validation analyses indicated that this was a robust method for Cq. linealis and Oc. vigilax. In contrast, trap counts for the container-breeding species Oc. notoscriptus were randomly distributed and the interpolated counts were not reliable. Comparisons of weekly contour maps of adult mosquito counts indicated a consistent spatial pattern for Oc. vigilax and Cq. linealis. Particular geographic areas had consistently high or low numbers of vectors, and these patterns were stable from year to year. Definition of geographic areas with consistently high or low numbers of vectors may allow control activities to be focused in areas with the greatest risk of arbovirus transmission.

Isolation of La Crosse, Cache Valley, and Potosi viruses from Aedes mosquitoes (Diptera: Culicidae) collected at used-tire sites in Illinois during 1994-1995.

Prospective studies were conducted at used-tire sites in Illinois during 1994-1995 in an effort to isolate arboviruses from mosquitoes, particularly Aedes albopictus (Skuse) and Aedes triseriatus (Say). Three isolates of Potosi virus were obtained from Ae. albopictus collected at a waste tire site in Jasper County during 1994 and 1995. Also, a single isolate of Cache Valley virus was obtained from Ae. albopictus collected at the Jasper County site during 1995. These are the first records of arbovirus isolations from Ae. albopictus in Illinois and the first isolate of Cache Valley virus from this mosquito species. During 1994, two isolates of La Crosse virus were made from Ae. triseriatus collected at a used-tire site in Peoria County in proximity to the residence of a human La Crosse encephalitis case. This is the first evidence in Illinois that indicates increased risk to humans living near used-tire sites, which may serve as foci for production of Ae. triseriatus, the vector of La Crosse virus. Tire removal and improved environmental sanitation at such sites may greatly reduce the abundance of vector mosquitoes, and, therefore, the risk of arbovirus transmission.

Mosquito-borne arboviruses in Australia: the current scene and implications of climate change for human health

Of the mosquito-borne arboviruses, the encephalitic Murray Valley encephalitis and Kunjin viruses are a major public health concern, but the arthritides Ross River and Barmah Forest viruses are more important in a public health sense, being responsible for a far greater number of infections. Reported cases of Ross River totalled approximately 30 000 during 1991–1996; there have been several widely separated outbreaks of Barmah Forest in recent years and case reports are increasing annually. Surveillance programmes have increased our understanding of the geographic regions, climatic conditions and vector factors associated with viruses. Virus activity is widespread but is often localised, is driven primarily by mosquito abundance and various species are involved; host factors are involved also, but are not well understood. Typically, mosquito populations are governed by availability of habitat and environmental conditions. Models of climate change predict increases in rainfall, tides and temperature for parts of Australia, and such changes have the potential to increase the risk of arbovirus transmission by increasing the distribution and abundance of vectors, and duration of mosquito and arbovirus seasons. However, the amplitude of climate change is uncertain and the ecology of arbovirus transmission is complex. It is likely that some areas will have increases in arbovirus activity and human infection with predicted climate change, but risk of increased transmission will vary with locality, vector, host and human factors.