Mosquito Virus Research Articles

Whole genome phylogenetic investigation of a West Nile virus strain isolated from a tick sampled from livestock in north eastern Kenya.

BackgroundWest Nile virus (WNV) has a wide geographical distribution and has been associated to cause neurological disease in humans and horses. Mosquitoes are the traditional vectors for WNV; however, the virus has also been isolated from tick species in North Africa and Europe which could be a means of introduction and spread of the virus over long distances through migratory birds. Although WNV has been isolated in mosquitoes in Kenya, paucity of genetic and pathogenicity data exists. We previously reported the isolation of WNV from ticks collected from livestock and wildlife in Ijara District of Kenya, a hotspot for arbovirus activity. Here we report the full genome sequence and phylogenetic investigation of their origin and relation to strains from other regions.MethodsA total of 10,488 ticks were sampled from animal hosts, classified to species and processed in pools of up to eight ticks per pool. Virus screening was performed by cell culture, RT-PCR and sequencing. Phylogenetic analysis was carried out to determine the evolutionary relationships of our isolate.ResultsAmong other viruses, WNV was isolated from a pool of Rhipicephalus pulchellus sampled from cattle, sequenced and submitted to GenBank (Accession number: KC243146). Comparative analysis with 27 different strains revealed that our isolate belongs to lineage 1 and clustered relatively closely to isolates from North Africa and Europe, Russia and the United States. Overall, Bayesian analysis based on nucleotide sequences showed that lineage 1 strains including the Kenyan strain had diverged 200 years ago from lineage 2 strains of southern Africa. Ijara strain collected from a tick sampled on livestock was closest to another Kenyan strain and had diverged 20 years ago from strains detected in Morocco and Europe and 30 years ago from strains identified in the USA.ConclusionTo our knowledge, this is the first characterized WNV strain isolated from R. pulchellus. The epidemiological role of this tick in WNV transmission and dissemination remains equivocal but presents tick verses mosquito virus transmission has been neglected. Genetic data of this strain suggest that lineage 1 strains from Africa could be dispersed through tick vectors by wild migratory birds to Europe and beyond.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-014-0542-2) contains supplementary material, which is available to authorized users.

Mosquito surveillance for prevention and control of emerging mosquito-borne diseases in Portugal - 2008-2014.

Mosquito surveillance in Europe is essential for early detection of invasive species with public health importance and prevention and control of emerging pathogens. In Portugal, a vector surveillance national program—REVIVE (REde de VIgilância de VEctores)—has been operating since 2008 under the custody of Portuguese Ministry of Health. The REVIVE is responsible for the nationwide surveillance of hematophagous arthropods. Surveillance for West Nile virus (WNV) and other flaviviruses in adult mosquitoes is continuously performed. Adult mosquitoes—collected mainly with Centre for Disease Control light traps baited with CO2—and larvae were systematically collected from a wide range of habitats in 20 subregions (NUTS III). Around 500,000 mosquitoes were trapped in more than 3,000 trap nights and 3,500 positive larvae surveys, in which 24 species were recorded. The viral activity detected in mosquito populations in these years has been limited to insect specific flaviviruses (ISFs) non-pathogenic to humans. Rather than emergency response, REVIVE allows timely detection of changes in abundance and species diversity providing valuable knowledge to health authorities, which may take control measures of vector populations reducing its impact on public health. This work aims to present the REVIVE operation and to expose data regarding mosquito species composition and detected ISFs.

Disease Vector Mapping Via Environmental, Climatological and Sociological Factors

The mosquito Aedes (Ae). aegypti serves as the primary urban vector for the virus that causes dengue, the most prevalent vector-borne viral disease on Earth. The probability of contracting dengue is modulated by climatic conditions that affect both Ae. aegypti development and the incubation period of dengue virus in adult mosquitoes, as well as socio-economic and human behavioral factors that enhance risk for vector-human contact and virus transmission. Vaccines or therapeutics are not yet available for use against dengue virus. Dengue virus infections often are asymptomatic, and there is an inherent delay between the onset date of dengue illness and the date the case is reported to the public health authorities. It is therefore critical to provide public health officials with a mechanism to better prepare in advance of an impending dengue epidemic. Here we describe a new decision support tool to inform public health planning and preparedness (e.g., the development of mosquito vector intervention strategies, garnering community awareness of dengue risk and support for control interventions). The tool, developed to be run in near-real-time, consists of a linkage of new and existing physically-based models that simulate 1) meteorological conditions over an area of interest; 2) the number and types of containers in a region (which are closely linked to socioeconomic and behavioral factors); 3) the temperature and volume of water in artificial container habitats in which Ae. aegypti develops; 4) the life cycle of the mosquito itself; and 5) the development of dengue virus and its transmission be of the mosquito itself; and 5) the development of dengue virus and its transmission between Ae. aegypti and humans.

Three-way interactions between mosquito population, viral strain and temperature underlying chikungunya virus transmission potential.

Interactions between pathogens and their insect vectors in nature are under the control of both genetic and non-genetic factors, yet most studies on mosquito vector competence for human pathogens are conducted in laboratory systems that do not consider genetic and/or environmental variability. Evaluating the risk of emergence of arthropod-borne viruses (arboviruses) of public health importance such as chikungunya virus (CHIKV) requires a more realistic appraisal of genetic and environmental contributions to vector competence. In particular, sources of variation do not necessarily act independently and may combine in the form of interactions. Here, we measured CHIKV transmission potential by the mosquito Aedes albopictus in all combinations of six worldwide vector populations, two virus strains and two ambient temperatures (20°C and 28°C). Overall, CHIKV transmission potential by Ae. albopictus strongly depended on the three-way combination of mosquito population, virus strain and temperature. Such genotype-by-genotype-by-environment (G × G × E) interactions question the relevance of vector competence studies conducted with a simpler set of conditions. Our results highlight the need to account for the complex interplay between vectors, pathogens and environmental factors to accurately assess the potential of vector-borne diseases to emerge.

Molecular epidemiology of Japanese encephalitis virus in mosquitoes in Taiwan during 2005-2012.

Japanese encephalitis (JE) is a mosquito-borne zoonotic disease caused by the Japanese encephalitis virus (JEV). Pigs and water birds are the main amplifying and maintenance hosts of the virus. In this study, we conducted a JEV survey in mosquitoes captured in pig farms and water bird wetland habitats in Taiwan during 2005 to 2012. A total of 102,633 mosquitoes were collected. Culex tritaeniorhynchus was the most common mosquito species found in the pig farms and wetlands. Among the 26 mosquito species collected, 11 tested positive for JEV by RT-PCR, including Cx. tritaeniorhynchus, Cx. annulus, Anopheles sinensis, Armigeres subalbatus, and Cx. fuscocephala. Among those testing positive, Cx. tritaeniorhynchus was the predominant vector species for the transmission of JEV genotypes I and III in Taiwan. The JEV infection rate was significantly higher in the mosquitoes from the pig farms than those from the wetlands. A phylogenetic analysis of the JEV envelope gene sequences isolated from the captured mosquitoes demonstrated that the predominant JEV genotype has shifted from genotype III to genotype I (GI), providing evidence for transmission cycle maintenance and multiple introductions of the GI strains in Taiwan during 2008 to 2012. This study demonstrates the intense JEV transmission activity in Taiwan, highlights the importance of JE vaccination for controlling the epidemic, and provides valuable information for the assessment of the vaccine's efficacy.

Monitoring of West Nile virus in mosquitoes between 2011-2012 in Hungary.

West Nile virus (WNV) is a widely distributed mosquito-borne flavivirus. WNV strains are classified into several genetic lineages on the basis of phylogenetic differences. Whereas lineage 1 viruses are distributed worldwide, lineage 2 WNV was first detected outside of Africa in Hungary in 2004. Since then, WNV-associated disease and mortality in animal and human hosts have been documented periodically in Hungary. After the first detection of WNV from a pool of Culex pipiens mosquitoes in 2010, samples were collated from several sources and tested in a 2-year monitoring program. Collection areas were located in the Southern Transdanubium, in northeastern Hungary, in eastern Hungary, and in southeastern Hungary. During the 2 years, 23,193 mosquitoes in 645 pools were screened for WNV virus presence with RT-PCR. Three pools were found positive for WNV in 2011 (one pool of Ochlerotatus annulipes collected in Fényeslitke in June, one pool of Coquillettidia richiardii collected in Debrecen, Fancsika-tó, in July, and one pool of Cx. pipiens captured near Red-Footed Falcon colonies at Kardoskút in September). The minimal infection rate (MIR=proportion of infected mosquitoes per 1000 mosquitoes) of all mosquito pools was 0.25, whereas the MIR of infected species was 2.03 for O. annulipes, 0.63 for C. richiardii, and 2.70 for C.x pipiens. Molecular data have demonstrated that the same lineage 2 WNV strain has circulated in wild birds, horses, humans, and mosquitoes in Hungary since 2004. Mosquito-based surveillance successfully complemented the ongoing, long-term passive surveillance system and it was useful for the early detection of WNV circulation.

Lack of evidence for the presence of Schmallenberg virus in mosquitoes in Germany, 2011.

BackgroundIn 2011, a novel orthobunyavirus of the Simbu serogroup was discovered near the German-Dutch border and named Schmallenberg virus (SBV). So far, SBV genome has been detected in various field-collected Culicoides species; however, other members of the Simbu serogroup are also transmitted by mosquitoes.FindingsIn the present study, approximately 50,000 mosquitoes of various species were collected during summer and early autumn 2011 in Germany. None of them tested positive in an SBV-specific real-time PCR.ConclusionsThe absence of SBV in mosquitoes caught in 2011 in Germany suggests that they play no or only a negligible role in the spread of the disease.

Molecular characterization of flaviviruses from field-collected mosquitoes in northwestern Italy, 2011-2012.

BackgroundThe genus Flavivirus comprises several mosquito-borne species, including the zoonotic pathogens West Nile and Usutu virus, circulating in animals and humans in Italy since 1998. Due to its ecological and geographical features, Piedmont is considered a risk area for flavivirus transmission. Here we report the results of a flavivirus survey (detection and genetic characterization) of mosquitoes collected in Piedmont in 2012 and the genetic characterization of three strains detected in 2011.MethodsPools of 1–203 mosquitoes, upon RNA extraction with TRIzol, were screened by a PCR assay for a 263 bp fragment of the Flavivirus NS5 gene. All positive samples were tested with a specific PCR for the E protein gene of Usutu virus and a generic Flavivirus RT-nested-PCR for a larger tract of the NS5 gene before sequencing. Phylogenetic trees were built with both NS5 fragments of representative Flavivirus species. DNA extracts of part of the positive pools were tested to detect sequences integrated in the host genome.ResultsThirty-four mosquito pools resulted positive for flaviviruses, and twenty-five flavivirus sequences underwent phylogenetic analysis for the short NS5 fragment. Among the 19 sequences correlating with the insect-specific flavivirus group, ten samples, retrieved from Aedes albopictus, clustered within Aedes flavivirus, while the other nine aggregated in a separate clade composed of strains from various mosquito species (mainly Aedes vexans) from Piedmont and the Czech Republic. Six out of these nine also presented a DNA form of the sequence. The remaining sequences belonged to the mosquito-borne group: four, all from Culex pipiens, correlated to Italian Usutu virus strains, whereas two, from Ochlerotatus caspius, were highly similar to Marisma mosquito virus (MMV).ConclusionsOur findings confirm the circulation of Usutu virus and of the potentially zoonotic Marisma mosquito virus in Piedmont. This is the first detection of Aedes flavivirus in Piedmont. Finally, further evidence for the integration of Flavivirus nucleic acid into the host genome has been shown. These results underline the importance of continuing intense mosquito-based surveillance in Piedmont, supported by a mosquito control program in areas at high risk for human exposure.

A spatial simulation model for dengue virus infection in urban areas.

BackgroundThe World Health Organization estimates that the global number of dengue infections range between 80–100 million per year, with some studies estimating approximately three times higher numbers. Furthermore, the geographic range of dengue virus transmission is extending with the disease now occurring more frequently in areas such as southern Europe. Ae. aegypti, one of the most prominent dengue vectors, is endemic to the far north-east of Australia and the city of Cairns frequently experiences dengue outbreaks which sometimes lead to large epidemics.MethodA spatially-explicit, individual-based mathematical model that accounts for the spread of dengue infection as a result of human movement and mosquito dispersion is presented. The model closely couples the four key sub-models necessary for representing the overall dynamics of the physical system, namely those describing mosquito population dynamics, human movement, virus transmission and vector control. Important features are the use of high quality outbreak data and mosquito trapping data for calibration and validation and a strategy to derive local mosquito abundance based on vegetation coverage and census data.ResultsThe model has been calibrated using detailed 2003 dengue outbreak data from Cairns, together with census and mosquito trapping data, and is shown to realistically reproduce a further dengue outbreak. The simulation results replicating the 2008/2009 Cairns epidemic support several hypotheses (formulated previously) aimed at explaining the large-scale epidemic which occurred in 2008/2009; specifically, while warmer weather and increased human movement had only a small effect on the spread of the virus, a shorter virus strain-specific extrinsic incubation time can explain the observed explosive outbreak of 2008/2009.ConclusionThe proof-of-concept simulation model described in this study has potential as a tool for understanding factors contributing to dengue spread as well as planning and optimizing dengue control, including reducing the Ae. aegypti vector population and for estimating the effectiveness and cost-effectiveness of future vaccination programmes. This model could also be applied to other vector borne viral diseases such as chikungunya, also spread by Ae. aegypti and, by re-parameterisation of the vector sub-model, to dengue and chikungunya viruses spread by Aedes albopictus.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2334-14-447) contains supplementary material, which is available to authorized users.

Correlating Remote Sensing Data with the Abundance of Pupae of the Dengue Virus Mosquito Vector, Aedes aegypti, in Central Mexico

Using a geographic transect in Central Mexico, with an elevation/climate gradient, but uniformity in socio-economic conditions among study sites, this study evaluates the applicability of three widely-used remote sensing (RS) products to link weather conditions with the local abundance of the dengue virus mosquito vector, Aedes aegypti (Ae. aegypti). Field-derived entomological measures included estimates for the percentage of premises with the presence of Ae. aegypti pupae and the abundance of Ae. aegypti pupae per premises. Data on mosquito abundance from field surveys were matched with RS data and analyzed for correlation. Daily daytime and nighttime land surface temperature (LST) values were obtained from Moderate Resolution Imaging Spectroradiometer (MODIS)/Aqua cloud-free images within the four weeks preceding the field survey. Tropical Rainfall Measuring Mission (TRMM)-estimated rainfall accumulation was calculated for the four weeks preceding the field survey. Elevation was estimated through a digital elevation model (DEM). Strong correlations were found between mosquito abundance and RS-derived night LST, elevation and rainfall along the elevation/climate gradient. These findings show that RS data can be used to predict Ae. aegypti abundance, but further studies are needed to define the climatic and socio-economic conditions under which the correlations observed herein can be assumed to apply.

Molecular epidemiology of Japanese encephalitis virus in mosquitoes during an outbreak in China, 2013.

Japanese encephalitis virus (JEV) can cause serious encephalitis and Culex mosquitoes are the primary vector. In 2013, a JE outbreak occurred in Shandong Province, China with 407 confirmed cases, including 11 deaths. An investigation on JEV in mosquitoes during the outbreak was conducted. A total of 14,719 mosquitoes were collected at 3 sites. For the 12,695 Culex tritaeniorhynchus mosquitoes, 88/201 pooled samples were positive by RT-PCR for the presence of the pre-membrane or envelope protein coding genes. The maximum likelihood estimates of JEV positive individuals per 1,000 vectors were 12.0, 7.2, and 6.0 in the 3 sites respectively with an overall estimate of 9.1. Phylogenetic analysis on these pre-membrane (n = 72) and envelope (n = 26) sequences with those of reference strains revealed they belonged to genotype I. This study describes the molecular epidemiology of JEV and suggests the high infection rate in mosquitoes is an important factor for the outbreak.

A comparison of three approaches to identify West Nile Virus mosquito space-time hotspots in the Houston Vicinity for the period 2002–2011

From 2002 to 2011, West Nile virus mosquitoes (WNV) has been ever-present in traps across Harris County, TX which contains the city of Houston. Disease-positive trap locations have peaked twice, from 2002 to 2006 and then again from 2009 onwards. This paper will examine fine scale spatial and temporal patterns in disease-positive mosquito traps for the Houston area across this time frame, using three different analytical approaches: kernel density, spatial filtering and SaTScan. The purpose of this paper is twofold. Firstly, to identify spatial and space-time clusters of WNV in order to spatially prioritize subsequent research for causative associations. Secondly, to compare the effectiveness of three methods that vary in complexity and ease of use in order to suggest a transferable methodology for mosquito control and environmental health departments across the United States with only lower level GIS skillsets. This paper also illustrates a successful ongoing academic and mosquito control collaboration with the Harris County Public Health Services Mosquito Control Division's (MCD) program.

Apropos: ‘Preliminary evaluation on the efficiency of the kit Platelia Dengue NS1 Ag-ELISA to detect dengue virus in dried Aedes aegypti: a potential tool to improve dengue surveillance’

Only simple, point-of-care, assay formats of the Platellia Dengue NS1 Ag-ELISA would be suitable to identify Dengue virus in Aedes aegypti mosquitoes in dengue-endemic areas lacking sophisticated laboratory infrastructure and trained laboratory personnel.

Whole genome phylogenetic investigation of a West Nile virus strain isolated from a tick sampled from livestock in north eastern Kenya

West Nile virus (WNV) has a wide geographical distribution and has been associated to cause neurological disease in humans and horses. Mosquitoes are the traditional vectors for WNV; however, the virus has also been isolated from tick species in North Africa and Europe which could be a means of introduction and spread of the virus over long distances through migratory birds. Although WNV has been isolated in mosquitoes in Kenya, paucity of genetic and pathogenicity data exists. We previously reported the isolation of WNV from ticks collected from livestock and wildlife in Ijara District of Kenya, a hotspot for arbovirus activity. Here we report the full genome sequence and phylogenetic investigation of their origin and relation to strains from other regions. A total of 10,488 ticks were sampled from animal hosts, classified to species and processed in pools of up to eight ticks per pool. Virus screening was performed by cell culture, RT-PCR and sequencing. Phylogenetic analysis was carried out to determine the evolutionary relationships of our isolate. Among other viruses, WNV was isolated from a pool of Rhipicephalus pulchellus sampled from cattle, sequenced and submitted to GenBank (Accession number: KC243146). Comparative analysis with 27 different strains revealed that our isolate belongs to lineage 1 and clustered relatively closely to isolates from North Africa and Europe, Russia and the United States. Overall, Bayesian analysis based on nucleotide sequences showed that lineage 1 strains including the Kenyan strain had diverged 200 years ago from lineage 2 strains of southern Africa. Ijara strain collected from a tick sampled on livestock was closest to another Kenyan strain and had diverged 20 years ago from strains detected in Morocco and Europe and 30 years ago from strains identified in the USA. To our knowledge, this is the first characterized WNV strain isolated from R. pulchellus. The epidemiological role of this tick in WNV transmission and dissemination remains equivocal but presents tick verses mosquito virus transmission has been neglected. Genetic data of this strain suggest that lineage 1 strains from Africa could be dispersed through tick vectors by wild migratory birds to Europe and beyond.

A divergent insect-specific flavivirus that causes overt cytopathic effect in a mosquito cell-line

The genus Flavivirus (Flaviviridae) includes over seven dozen known enveloped viruses with ssRNA genomes, the evolution, geographic dispersion and epidemiology of which seems to have been shaped both by environmental constraints (involving ecological factors and human activities), and by their replication in vertebrate and invertebrate hosts. Most bona-fide flaviviruses have been classified as mosquito or tick-borne viruses, although for some no known invertebrate vector has yet been found. Over time, a divergent cluster including viral agents that seem to be specific of insect (Insect-Specific Flaviviruses-ISF) has been associated with the genus. The classification of ISF as flaviviruses is supported by their genomic organization, protein hydropathy plots, conserved polyprotein cleavage sites and enzyme domains, which are similar to those of classical flaviviruses. However, they are antigenically distinct, and share approximately the same level of nucleotide sequence identity with other members of the genus, as when compared to the members of the two other genera in the Flaviviridae family.This work reports the initial characterization, including near full-length sequence analysis, of an ISF which seems to form a divergent lineage within the ISF radiation (Figure 1), as indicated by phylogenetic analysis based on ac multiple sequence alignment of the NS5 flavivirus coding sequence. This virus, tentatively designated OCFV (Ochlerotatus caspius flavivirus) was isolated from clarified macerates of a pool of Aedes (Ochlerotatus) caspius (Pallas, 1771) adult mosquitoes (lab code number 174) using the Ae. albopictus-derived C6/36 cell line. The mosquitoes from which OCFV was isolated were collected using CDC-traps baited with CO2, and are amongst those found in high densities in the coastal, and estuarine, districts of Setúbal and Faro. Detection of viral genomes by RT-PCR using RNA directly extracted from supernatants of C6/36 infected cultures, collected 24h post-infection, indicated that the virus replicates rapidly in these cells. Nevertheless, but as expected for an ISF, it does not replicate in Vero cells. Unexpectedly, unlike most ISF OCFV seems to cause overt cytopathic effect in C6/36 cells, which rapidly detach from a solid support and round-up soon (24-48h) after infection. Electron-microscope analysis of thin-sections of C6/36 cells at 48h post-infection with OCFV revealed nuclear hyperplasy, and an evident enlargement of the intracisternal space of the nuclear envelope, which is filled by multiple sized vesicles. In some cells, a complex network of apparent membrane trabeculae was also clearly evident (Figure 2A). Viral particles seem to assemble at the endoplasmatic reticulum, from which viruses reach the cell surface by vesicular transport (Figure 2B). Unexpectedly for a flavivirus, but as previously observed, viral particles also seem to gemulate directly from the cytoplasmic membrane (Figure 2C).

Cell Phone-Based System (Chaak) for Surveillance of Immatures of Dengue Virus Mosquito Vectors

Capture of surveillance data on mobile devices and rapid transfer of such data from these devices into an electronic database or data management and decision support systems promote timely data analyses and public health response during disease outbreaks. Mobile data capture is used increasingly for malaria surveillance and holds great promise for surveillance of other neglected tropical diseases. We focused on mosquito-borne dengue, with the primary aims of: 1) developing and field-testing a cell phone-based system (called Chaak) for capture of data relating to the surveillance of the mosquito immature stages, and 2) assessing, in the dengue endemic setting of Mérida, Mexico, the cost-effectiveness of this new technology versus paper-based data collection. Chaak includes a desktop component, where a manager selects premises to be surveyed for mosquito immatures, and a cell phone component, where the surveyor receives the assigned tasks and captures the data. Data collected on the cell phone can be transferred to a central database through different modes of transmission, including near-real time where data are transferred immediately (e.g., over the Internet) or by first storing data on the cell phone for future transmission. Spatial data are handled in a novel, semantically driven, geographic information system. Compared with a pen-and-paper-based method, use of Chaak improved the accuracy and increased the speed of data transcription into an electronic database. The cost-effectiveness of using the Chaak system will depend largely on the up-front cost of purchasing cell phones and the recurring cost of data transfer over a cellular network.

Replacing a Native Wolbachia with a Novel Strain Results in an Increase in Endosymbiont Load and Resistance to Dengue Virus in a Mosquito Vector

Wolbachia is a maternally transmitted endosymbiotic bacterium that is estimated to infect up to 65% of insect species. The ability of Wolbachia to both induce pathogen interference and spread into mosquito vector populations makes it possible to develop Wolbachia as a biological control agent for vector-borne disease control. Although Wolbachia induces resistance to dengue virus (DENV), filarial worms, and Plasmodium in mosquitoes, species like Aedes polynesiensis and Aedes albopictus, which carry native Wolbachia infections, are able to transmit dengue and filariasis. In a previous study, the native wPolA in Ae. polynesiensis was replaced with wAlbB from Ae. albopictus, and resulted in the generation of the transinfected “MTB” strain with low susceptibility for filarial worms. In this study, we compare the dynamics of DENV serotype 2 (DENV-2) within the wild type “APM” strain and the MTB strain of Ae. polynesiensis by measuring viral infection in the mosquito whole body, midgut, head, and saliva at different time points post infection. The results show that wAlbB can induce a strong resistance to DENV-2 in the MTB mosquito. Evidence also supports that this resistance is related to a dramatic increase in Wolbachia density in the MTB's somatic tissues, including the midgut and salivary gland. Our results suggests that replacement of a native Wolbachia with a novel infection could serve as a strategy for developing a Wolbachia-based approach to target naturally infected insects for vector-borne disease control.

First indigenous transmission of Japanese Encephalitis in urban areas of National Capital Territory of Delhi, India

Until 2010, no Japanese encephalitis (JE) had been reported from Delhi. Upon report of four confirmed cases of JE in September 2011, detailed investigations were carried out to determine whether the cases were imported or indigenous. Entomological surveys were carried out and all mosquito pools were tested for the detection of JE virus by ELISA method using specific monoclonal antibody. Human blood samples from contacts of the patients were tested by IgM-captured ELISA method. Pig's blood samples were also tested for the detection of JE virus. Culex tritaeniorhynchus, Culex vishnui and Culex pseudovishnui mosquitoes were found. In contrast to rural areas, their breeding habitats were different in the city. 19 pools were tested. JE virus was detected in two pools of Cx.tritaeniorhynchus females reared from field-collected larvae, indicating vertical transmission. One pool of Cx.vishnui was also positive. This is the first report for the detection of JE virus in mosquitoes from Delhi. JE IgM antibodies in five contacts/residents indicate recent infection. JE virus was also detected in pigs. Present analysis shows that of four reported JE cases, three were confirmed indigenous, indicating that the virus is multiplying in the city. Mapping of infected JE vector mosquitoes in the cities is required for preventive measures to contain further spread of the disease.

Transmission dynamics of West Nile virus in mosquitoes and corvids and non-corvids

There are more than 300 avian species that can transmit West Nile virus (WNv). In general, the corvid and non-corvid families of birds have different responses to the virus, with corvids suffering a higher disease-induced mortality rate. By taking both corvids and non-corvids as the primary reservoir hosts and mosquitoes as vectors; we formulate and study a system of ordinary differential equations to model a single season of the transmission dynamics of WNv in the mosquito-bird cycle. We calculate the basic reproduction number and analyze the existence and stability of the equilibria. The existence of a backward bifurcation gives a further sub-threshold condition beyond the basic reproduction number for the spread of the virus. We also discuss the role of corvids and non-corvids in spreading the virus. We conclude that knowledge of the relative abundance of corvid bird species and other mammals assist us in accurate estimation of the epidemic of WNv.

Mosquito and West Nile virus surveillance in northeast Montana, U.S.A., 2005 and 2006

Mosquito and West Nile virus (WNV) surveillance was conducted on a national wildlife refuge in northeast Montana in 2005 and 2006, during which outbreaks of WNV in a colony of American white pelicans (Pelecanus erythrorhynchos Gmelin) (Pelecaniformes: Pelecanidae) resulted in juvenile mortality rates of ∼ 31%. Both years, floodwater species Ochlerotatus dorsalis (Meigen) (Diptera: Culicidae), Aedes vexans (Meigen) (Diptera: Culicidae) and Ochlerotatus flavescens (Muller) (Diptera: Culicidae) comprised 78% of the total collection and heightened host-seeking activity was observed from mid-June to mid-July. Culex tarsalis Coquillett (Diptera: Culicidae) was most active from mid-July to mid-August and comprised 18% of the collection in 2005 and 20% in 2006. However, fewer than 10% of the Cx. tarsalis females collected in 2006 were obtained adjacent to the pelicans' nesting grounds. Minimum infection rates per 1000 Cx. tarsalis tested for WNV were 1.36 in 2005 and 1.41 in 2006. All pools in which WNV was detected in 2006 were composed of females collected 10 km from the nesting grounds. Substantial juvenile pelican mortality in 2006 despite reductions in the population of the primary vector and in mosquito infection rates near the colony suggests that the methods used to detect the introduction of WNV were too coarse and that amplification of the virus within the colony may reflect causes other than mosquito infection.