Vector competence of mosquitoes (Diptera: Culicidae) from Maroochy Shire, Australia, for Barmah Forest virus

Mosquitoes were collected in light traps from Maroochy Shire and fed on blood containing the sympatric BF1611 strain of Barmah Forest virus (BF). Saltmarsh Aedes vigilax (Skuse) and freshwater Aedes procax (Skuse) were highly susceptible to infection, with ID50s of 10(1.7) and 10(1.5) African green monkey kidney (Vero) cell culture infectious dose, 50% endpoint (CCID50) per mosquito, respectively, followed by Aedes multiplex (Theobald) and Aedes funereus (Theobald) with 10(2.5) and 10(3.2) CCID50 per mosquito, respectively. Culex australicus Dobrotworsky & Drummond and Mansonia uniformis (Theobald) that were fed 10(3.6) CCID50 (Vero) BF per mosquito had infection rates of 28 and 60%, respectively. Only 8% of freshwater Culex annulirostris Skuse fed the same viral dose were infected. Evidence of virus transmission to mice was found with Ae. vigilax and Ae. procax, with transmission rates of 65 and 88% at 11-12 d after infection, respectively. Based on adult abundance, susceptibility to infection, and efficiency of virus transmission, Ae. vigilax would appear to be the most important vector of BF in Maroochy Shire.     

Spatial and temporal analysis of Ross River virus disease patterns at Maroochy Shire, Australia: association between human morbidity and mosquito (Diptera: Culicidae) abundance.

Notifications of Ross River (RR) virus disease in Maroochy Shire were mapped according to the patient's place of residence, and standardized morbidity ratios were calculated for each of 11 census districts, with 4 areas having higher than average overall rates of RR virus disease notification. Temporal analysis of RR virus disease notifications from each of the 11 areas indicated that epidemics of RR virus disease either were widespread, resulting in higher than average numbers of cases from the majority of areas, or limited to only certain areas. Mosquitoes were collected in CO2 light traps, and the numbers analyzed to determine whether there was any association between mosquito abundance and disease incidence. Seasonal light trap indices for the fresh water-breeding Culex annulirostris Skuse and brackish water-breeding Aedes funereus (Theobald) were correlated positively with disease incidence. There was no apparent association between Aedes vigilax (Skuse) or Aedes procax (Skuse) light trap indices and disease incidence. Identification of human subpopulations with higher than average rates of RR virus disease, and the mosquito vectors associated with virus transmission, should provide a framework for the development of focused and therefore more effective control programs.     

Vector competence of Australian mosquitoes (Diptera: Culicidae) for Japanese encephalitis virus

Australian mosquitoes were evaluated for their ability to become infected with and transmit a Torres Strait strain of Japanese encephalitis virus. Mosquitoes, which were obtained from either laboratory colonies and collected using Centers for Disease Control and Prevention light traps baited with CO2 and octenol or reared from larvae, were infected by feeding on a blood/sucrose solution containing 10(4.5 +/- 0.1) porcine stable-equine kidney (PS-EK) tissue culture infectious dose50/mosquito of the TS3306 virus strain. After 14 d, infection and transmission rates of 100% and 81%, respectively, were obtained for a southeast Queensland strain of Culex annulirostris Skuse, and 93% and 61%, respectively, for a far north Queensland strain. After 13 or more days, infection and transmission rates of > 90% and > or = 50%, respectively, were obtained for southeast Queensland strains of Culex sitiens Wiedemann and Culex quinquefasciatus Say, and a far north Queensland strain of Culex gelidus Theobald. Although infection rates were > 55%, only 17% of Ochlerotatus vigilax (Skuse) and no Cx. quinquefasciatus, collected from far north Queensland, transmitted virus. North Queensland strains of Aedes aegypti L., Ochlerotatus kochi (D?nitz), and Verrallina funerea (Theobald) were relatively refractory to infection. Vertical transmission was not detected among 673 F1 progeny of Oc. vigilax. Results of the current vector competence study, coupled with high field isolation rates, host feeding patterns and widespread distribution, confirm the status of Cx. annulirostris as the major vector of Japanese encephalitis virus in northern Australia. The relative roles of other species in potential Japanese encephalitis virus transmission cycles in northern Australia are discussed.     

Vector competence of Aedes aegypti, Culex sitiens, Culex annulirostris, and Culex quinquefasciatus (Diptera: Culicidae) for Barmah Forest virus

Aedes aegypti (L.), Culex sitiens Weidemann, Culex annulirostris Skuse, and Culex quinquefasciatus Say mosquitoes colonized at the Queensland Institute of Medical Research, Brisbane Australia, were fed on blood containing Barmah Forest virus (BF). Only Cx. annulirostris was susceptible to infection, with a median cell culture infectious dose (CCID50) of 10(3.36) per mosquito. Ae. aegypti and Cx. quinquefasciatus were infected experimentally, but at rates of < 9%. Cx. sitiens did not become infected. Infection rates for Cx. annulirostris fed 10(3.5) CCID50 of virus per mosquito, varied from 9 to 50% between 2 and 13 d after infection. Virus transmission to suckling mice by Cx. annulirostris occurred from 2 d after infection. Transmission of BF virus by Cx. annulirostris was 10% at 2 d after infection and did not exceed 8% thereafter. Although Cx. annulirostris may be infected and is able to transmit BF virus to suckling mice, it is nonetheless a relatively inefficient vector of the virus.     

Mosquito species (Diptera: Culicidae) and the transmission of Ross River virus in Brisbane, Australia

This study aimed to identify the major mosquito vectors of Ross River virus (family Togaviridae, genus Alphavirus, RRV) and to explore the threshold of mosquito abundance necessary for RRV transmission in Brisbane, Australia. Data on the monthly counts of RRV cases by statistical local areas from the Queensland Health and the monthly mosquito abundance in Brisbane between November 1998 and December 2001 from the Brisbane City Council were used to assess the pairwise relationship between mosquito abundance and the incidence of RRV disease over a range of time lags using cross-correlations. We used time series Poisson regression models to identify major mosquito species associated with incidence of RRV after adjusting for overdispersion, maximum temperature, autocorrelation, and seasonality. Our results show that Aedes vigilax (Skuse) (relative risk [RR] = 1.32; 95% CI = 1.01-1.74 per 100 mosquitoes per trap) and Culex annulirostris (Skuse) (RR = 1.14, 95% CI = 1.04-1.24 per 100 mosquitoes per trap) were most strongly associated with RRV transmission at a lag of 1 mo. Classification and regression tree (CART) analyses indicate that the occurrence of RRV was associated with an average monthly mosquito abundance ofAedes vigilax above 72 and Cx. annulirostris above 52. The validation analyses indicate that the crude agreement between predicted values and actual observations was 76% (sensitivity, 61%; specificity, 80%). The results may have applications as a decision support tool in planning disease control and risk-management programs.     

Experimental infection and transmission of Barmah Forest virus by Aedes vigilax (Diptera: Culicidae)

Aedes vigilax (Skuse) mosquitoes colonized from Townsville, Queensland, Australia, were fed on blood containing Barmah Forest virus (BF) isolated from the same species. The colony was susceptible to infection, with an ID50 of 10(2.6) CCID50 per mosquito. Infection and transmission rates for mosquitoes fed 10(3.5) CCID50 virus per mosquito varied from 58 to 100% and 36 to 100%, respectively, between days 3 and 13 after infection. Titers in infected mosquitoes were high by 5 d after infection and did not vary substantially thereafter. Virus transmission to suckling mice occurred from 3 d after infection. This work establishes that Ae. vigilax is a competent vector of BF.     

Isolation of arboviruses from mosquitoes (Diptera: Culicidae) collected from the Gulf Plains region of northwest Queensland,Australia

As part of investigations into Japanese encephalitis (JE) virus and related flaviviruses in northern Australia, 153,529 mosquitoes were collected and processed for virus isolation from the Gulf Plains region of northwest Queensland. Collections from within 30 km of each of the townships of Croydon, Normanton and Karumba yielded 3,087 (2.0%), 66,009 (43.0%), and 84,433 (55.0%) mosquitoes, respectively, from which 16 viruses were isolated. Four isolates of Murray Valley encephalitis (MVE), two of Kunjin (KUN), three of Ross River (RR), and one of Sindbis (SIN) viruses were obtained from Culex sitiens subgroup mosquitoes. Molecular identification of the mosquito species composition of these virus positive pools revealed that most isolates were from pools containing mainly Culex annulirostris Skuse and low numbers of Culex palpalis (Taylor). Only three pools, one each of MVE, KUN, and RR, were from mosquitoes identified exclusively as Cx. annulirostris. Other viruses isolated include one Edge Hill virus from Ochlerotatus normanensis (Taylor), an isolate of SIN from Anopheles meraukensis Venhuis, two isolates of RR from Anopheles amictus Edwards, and single isolates of RR from Anopheles bancroftii Giles andAedes lineatopennis (Ludlow). The isolate of RR from Ae. lineatopennis was the first reported from this species. The public health implications of these isolations in the Gulf Plains region are discussed briefly.     

Spatial dispersion of adult mosquitoes (Diptera: Culicidae) in a sylvatic focus of Venezuelan equine encephalitis virus.

We studied the spatial localization of mosquitoes in sylvatic focus of Venezuelan equine encephalitis virus in western Venezuela to identify mosquito species potentially involved in the hypothesized transport of viruses out of enzootic foci. The following criteria were used to identify species with potential for virus export: (1) common in the forest and surrounding area, (2) feeding on a wide range of vertebrates; (3) long dispersal capabilities, and (4) established vectorial competence for enzootic or epizootic VEE viruses. CDC traps baited with light/CO2 were operated for four and 12-h intervals to collect mosquitoes at four stations along two forest/open area transects from September to November 1997. We collected 60,444 mosquitoes belonging to 11 genera and 34 species. The most common species were Aedes serratus (Theobald), Ae. scapularis (Rondani), Ae. fulvus (Wiedmann), Culex nigripalus Theobald, Cx, (Culex) "sp", Cx. mollis Dyar & Knab, Cx. spissipes (Theobald), Cx. pedroi Sirivanakarn and Belkin, Psorophora ferox (Humboldt), Ps. albipes (Theobald), and Ps. cingulata (F.). Very few mosquitoes were captured during the (day in the open area outside the forest, suggesting that any virus export from the forest may occur at night. The following mosquitoes seemed to be mostly restricted to the forest habitat: Ae. serratus, Ps. ferox, Ps. albipes, sabethines, Cx. spissipes, Cx. pedroi, Cx. dunni Dyar, and Ae. fulvus. The main species implicated its potential virus export were Cx. nigripalpus, Ae. scapularis, and Mansonia titillans (Walker).     

Vector competence of selected African mosquito (Diptera: Culicidae) species for Rift Valley fever virus

Outbreaks of Rift Valley fever (RVF) in Egypt, Yemen, and Saudi Arabia have indicated the potential for this disease to spread from its enzootic areas in sub-Saharan Africa. Because little is known about the potential for most African mosquito species to transmit RVF virus (family Bunyaviridae, genus Phlebovirus, RVFV), we conducted studies to determine the vector competence of selected African species of mosquitoes for this virus. All eight species tested [Aedes palpalis (Newstead), Aedes mcintoshi Huang, Aedes circumluteolus (Theobald), Aedes calceatus Edwards, Aedes aegypti (L.), Culex antennatus (Becker), Culex pipiens (L.), and Culex quinquefasciatus Say], were susceptible to infection, and all except Ae. calceatus, Ae. aegypti and Cx. quinquefasciatus transmitted RVFV by bite after oral exposure. Estimated transmission rates for mosquitoes that successfully transmitted RVFV by bite ranged from 5% for Ae. mcintoshi to 39% for Ae. palpalis for mosquitoes that fed on a hamster with a viremia > or = 10(8) plaque-forming units of virus/ml. We did not recover RVFV from any of 3,138 progeny of infected female mosquitoes. RVFV is unusual among arboviruses in that it has been isolated in nature from a large number of species and that numerous mosquitoes and other arthropods are able to transmit this virus in the laboratory. The recent introduction and spread of West Nile virus into the Americas and the spread of RVFV to the Arabian Peninsula illustrates the potential for viruses, once enzootic in Africa, to spread to other parts of the world.     

Ross River virus disease clusters and spatial relationship with mosquito biting exposure in Redland Shire, southern Queensland, Australia

The spatial heterogeneity in the risk of Ross River virus (family Togaviridae, genus Alphavirus, RRV) disease, the most common mosquito-borne disease in Australia, was examined in Redland Shire in southern Queensland, Australia. Disease cases, complaints from residents of intense mosquito biting exposure, and human population data were mapped using a geographic information system. Surface maps of RRV disease age-sex standardized morbidity ratios and mosquito biting complaint morbidity ratios were created. To determine whether there was significant spatial variation in disease and complaint patterns, a spatial scan analysis method was used to test whether the number of cases and complaints was distributed according to underlying population at risk. Several noncontiguous areas in proximity to productive saline water habitats of Aedes vigilax (Skuse), a recognized vector of RRV, had higher than expected numbers of RRV disease cases and complaints. Disease rates in human populations in areas which had high numbers of adult Ae. vigilax in carbon dioxide- and octenol-baited light traps were up to 2.9 times those in areas that rarely had high numbers of mosquitoes. It was estimated that targeted control of adult Ae. vigilax in these high-risk areas could potentially reduce the RRV disease incidence by an average of 13.6%. Spatial correlation was found between RRV disease risk and complaints from residents of mosquito biting. Based on historical patterns of RRV transmission throughout Redland Shire and estimated future human population growth in areas with higher than average RRV disease incidence, it was estimated that RRV incidence rates will increase by 8% between 2001 and 2021. The use of arbitrary administrative areas that ranged in size from 4.6 to 318.3 km2 has the potential to mask any small scale heterogeneity in disease patterns. With the availability of georeferenced data sets and high-resolution imagery, it is becoming more feasible to undertake spatial analyses at relatively small scales.     

Role of Verrallina funerea (Diptera: Culicidae) in transmission of Barmah Forest virus and Ross River virus in coastal areas of eastern Australia

Verrallina funerea (Theobald) (Diptera: Culicidae) is a brackish water mosquito species found most commonly in Indonesia, Papua New Guinea, and the northeastern coastal regions of Australia. Aspects of the vector competence of this species for Barmah Forest virus (family Togaviridae, genus Alphavirus, BFV) and Ross River virus (family Togaviridae, genus Alphavirus, RRV), two medically important arboviruses in Australia, were investigated. Laboratory-reared Ve. funerea were moderately susceptible to experimental infection with BFV (median cell culture infectious dose of 10(3.6) per mosquito) and were capable of transmission to suckling mice (52% after a 9-12-d extrinsic incubation period). Maximum salivary gland infections for BFV (65%) and RRV (50%) were observed 8 and 10 d postinfection, respectively. To examine any regional differences in vector competence, field populations (separated by up to 200 km) of Ve. funerea and Aedes vigilax (Skuse) from southeastern Queensland and northern New South Wales were fed BFV and RRV, and subsequent infection rates were compared. For both viruses, no statistically significant variations in body, disseminated, or salivary gland infection rates were found in either Ve. funerea or Ae. vigilax. The results from this study indicate that Ve. funerea may have an important role as an amplification vector during outbreaks of both viruses and that local government authorities should rapidly treat brackish water habitats to control this species during periods of increased disease activity.     

Survey of adult mosquitoes (Diptera: Culicidae) during irrigation development in the Mahaweli Project, Sri Lanka.

A survey of adult mosquitoes was done during 1986-1987 in an area undergoing irrigation development in the Mahaweli Project of Sri Lanka. The study encompassed two 12-mo phases of settlement and irrigation, respectively. Diurnal human bait catches yielded 1,427 female mosquitoes in 27 species. Nocturnal collections (human bait, CDC light traps, indoor resting catches) produced a total of 30,491 females in 67 species. Comparisons with an earlier survey in the predevelopment forest showed that the diurnal human biting abundance of Aedes albopictus (Skuse) remained stable, whereas the abundance of Ae. novalbopictus Barraud and Ae. w-albus Theobald declined drastically after development. Man-mosquito contact increased substantially for nocturnally active Anopheles annularis van der Wulp, An. subpictus Grassi, Mansonia uniformis (Theobald), Culex tritaeniorhynchus Giles, and Cx. quinefasciatus Say and decreased for Ma. annulifera (Theobald) and Cx. gelidus Theobald. A small increase occurred in An. culicifacies Giles, An. vagus Donitz, An. varuna lyengar, Cx. fuscocephala Theobald, and Cx. whitmorei (Giles); Cx. pseudovishnui Colless decreased. The effect of changes in mosquito abundance on the increased transmission of malaria and Japanese encephalitis is discussed.     

Vector competence of North American mosquitoes (Diptera: Culicidae) for West Nile virus

We evaluated the potential for several North American mosquito species to transmit the newly introduced West Nile (WN) virus. Mosquitoes collected in the New York City metropolitan area during the recent WN virus outbreak, at the Assateague Island Wildlife Refuge, VA, or from established colonies were allowed to feed on chickens infected with WN virus isolated from a crow that died during the 1999 outbreak. These mosquitoes were tested approximately 2 wk later to determine infection, dissemination, and transmission rates. Aedes albopictus (Skuse), Aedes atropalpus (Coquillett), and Aedes japonicus (Theobald) were highly susceptible to infection, and nearly all individuals with a disseminated infection transmitted virus by bite. Culex pipiens L. and Aedes sollicitans (Walker) were moderately susceptible. In contrast, Aedes vexans (Meigen), Aedes aegypti (L.), and Aedes taeniorhynchus (Wiedemann) were relatively refractory to infection, but individual mosquitoes inoculated with WN virus did transmit virus by bite. Infected female Cx. pipiens transmitted WN virus to one of 1,618 F1 progeny, indicating the potential for vertical transmission of this virus. In addition to laboratory vector competence, host-feeding preferences, relative abundance, and season of activity also determine the role that these species could play in transmitting WN virus.     

Barmah Forest virus epidemic on the south coast of New South Wales, Australia, 1994-1995: viruses, vectors, human cases, and environmental factors

In 1995, the largest recorded outbreak of human disease resulting from infection with the mosquito transmitted alphavirus Barmah Forest (BF) virus occurred along the south coast of New South Wales, Australia. The virus was first isolated in early January from mosquitoes collected at Batemans Bay and predisposed the recognition of 135 human clinical cases. The cases of BF virus were identified initially from Batemans Bay during late January, and the majority (30%) of all cases came from this town. After 5 wk, all major centers on the south coast had clinical patients. Aedes vigilax (Skuse) were especially abundant at Batemans Bay, with levels up to 8 times greater than normal. This species yielded 111 isolates and appeared to be the major vector of BF virus. Attempts to examine if BF virus was maintained in the field by vertical transmission within Ae. vigilax populations were unsuccessful; no evidence of vertical transmission with BF virus, nor any other arbovirus, was found in > 17,000 adults emerging from field-collected larvae from the region following peak virus activity. In addition to BF virus, other viruses were recovered from field-collected adult mosquitoes, including Ross River (10 isolates), Edge Hill (21), and Stratford (10). Ae. vigilax again yielded the majority of these viral isolates. The BF virus outbreak appeared to be associated with several factors. A lack of recent BF virus activity in the region provided a highly susceptible human population, and unusual weather conditions of above average rainfall coupled with high tides resulted in extraordinarily large populations of Ae. vigilax.     

Definition of species in the Culex sitiens subgroup (Diptera: Culicidae) from Papua New Guinea and Australia

Japanese encephalitis is a disease of major importance in Asia, which from 1995 has extended its range into the north of Australia. Culex annulirostris Skuse of the Culex sitiens subgroup of Culex was incriminated as the most likely mosquito vector. Because misidentifications previously have occurred within this subgroup, our paper defines electrophoretic methods to identify Culex specimens from Queensland and the Northern Territory in Australia and from Western Province in Papua New Guinea. In collections from 1996 to 1998 Culex sitiens Wiedemann, Cx. palpalis Taylor, and Cx. annulirostris comprised 93.1% of the total banded proboscis Culex mosquitoes collected; 6.9% remained unidentified to species by morphological or electrophoretic examination. The prevalence of Cx. palpalis was variable and appeared to be associated with below average rainfall during 1997. In Papua New Guinea, this species comprised up to 97.4% of banded proboscis Culex, with smaller numbers collected from the Torres Strait Islands and Cape York; none was collected from other Queensland localities. Cx. annulirostris was more prevalent during years of average or above average rainfall and was predominant in most localities, especially in the Torres Strait and Queensland. These data confirmed the problems in separating Cx. sitiens from Cx. palpalis and Cx. annulirostris using morphological characters.     

Vector competence of rural and urban strains of Aedes (Stegomyia) albopictus (Diptera: Culicidae) from São Paulo State, Brazil for IC, ID, and IF subtypes of Venezuelan equine encephalitis virus

Aedes albopictus (Skuse) is an Asiatic mosquito species that has spread and colonized all continents except Antarctica. It has major public health importance because it is a potential vector of several pathogens. The objectives of our study were to analyze the vector competence of urban and rural strains of Ae. albopictus from S?o Paulo State (Brazil) for Venezuelan equine encephalitis virus (VEE) subtypes IC, ID, and IF, and to evaluate the effect of infection with subtype IC of VEE on mosquito longevity. Both mosquito strains were susceptible to subtypes IC and ID, but the infection rate for subtype IF was low. Infection and transmission rates of Ae. albopictus for subtype IC were similar to those reported for Ochlerotatus taeniorhynchus (Wiedemann). The high infection, dissemination, and transmission rates for subtype ID reported for Oc. fulvus (Wiedemann) and Culex (Melanoconion) spp. are comparable with those found in this study. We found significant differences in the susceptibility to subtype IC between rural and urban populations of S?o Paulo. Significant survival rate differences were observed between uninfected and infected mosquitoes, but there were no differences in survival between rural and urban mosquito strains.     

Habitat segregation of mosquito arbovirus vectors in south Florida

Oviposition traps set in rural to urban environments in three south Florida counties were colonized predominantly by Culex quinquefasciatus Say (35.1%), Aedes albopictus (Skuse) (34.5%), Aedes aegypti (L.) (23.8%), and Culex nigripalpus Theobald (6.6%) during 1 yr of monthly sampling. Significant differences were detected among counties for abundances of Cx. quinquefasciatus and for percentage composition of that species and Ae. albopictus. Aerial images of habitats around each collection site were digitized, and coverages by each of 16 habitat variables were recorded. Abundances ofAe. aegypti were positively related to habitat variables associated with urbanization and negatively correlated to those reflecting rural characteristics. Multiple regression models of habitat selection explained similar proportions of variances in abundance of Ae. aegypti and Ae. albopictus, but signs of significant variables were opposite for these two species. No consistent trends of habitat associations were observed among counties for the two Culex spp. Co-occurrences of the four species in individual traps depended on container type (tub versus cup), and, for Aedes spp. with Culex spp., county. The results underscore the importance of scale in evaluating habitat selection and the utility of quantifiable habitat characteristics of intermediate scale to identify site characteristics favored by the arboviral vectors Ae. aegypti and Ae. albopictus.