Blood-feeding patterns of the Culex pipiens complex in Sacramento and Yolo Counties, California

Mosquitoes in the Culex pipiens complex are competent vectors of West Nile virus (WNV; family Flaviviridae, genus Flavivirus) in the laboratory, and field-collected mosquitoes have tested positive for the virus in California and elsewhere. A better understanding of Cx. pipiens complex blood-feeding patterns will help define the threat that these mosquitoes pose to human health and their role in WNV amplification in northern California. We collected blood-engorged Cx. pipiens complex mosquitoes from resting sites near and away from human habitation in Sacramento and Yolo Counties. Cytochrome c oxidase 1 gene sequences were used to identify the vertebrate species from which blood meals were taken. Of 330 engorged mosquitoes collected at 28 sites from June through August 2007 and May through August 2008, >99% fed on an avian host. Three mosquitoes contained bovine blood and none had fed on a human. American Robins (Turdus migratorius) were bitten most often, and the proportion of American Robin blood meals increased significantly over the summer. Other important avian hosts included House Finches (Carpodacus mexicanus), Barn Swallows (Hirundo rustica), Western Meadowlarks (Sturnella neglecta), and Mourning Doves (Zenaida macroura). In rural areas, Barn Swallows, Brewer's Blackbirds (Euphagus cyanocephalus), and House Sparrows (Passer domesticus) were frequent hosts. In settings near human habitation, Mourning Doves and Western Meadowlarks were common hosts. Our data indicate that in north central California mosquitoes in the Cx. pipiens complex may be more important as epiornitic than epidemic vectors of WNV.     

Spatial variation in host feeding patterns of Culex tarsalis and the Culex pipiens complex (Diptera: Culicidae) in California

West Nile virus (family Flaviviridae, genus Flavivirus, WNV) is now endemic in California across a variety of ecological regions that support a wide diversity of potential avian and mammalian host species. Because different avian hosts have varying competence for WNV, determining the blood-feeding patterns of Culex (Diptera: Culicidae) vectors is a key component in understanding the maintenance and amplification of the virus as well as tangential transmission to humans and horses. We investigated the blood-feeding patterns of Culex tarsalis Coquillett and members of the Culex pipiens L. complex from southern to northern California. Nearly 100 different host species were identified from 1,487 bloodmeals, by using the mitochondrial gene cytochrome c oxidase I (COI). Cx. tarsalis fed on a higher diversity of hosts and more frequently on nonhuman mammals than did the Cx. pipiens complex. Several WNV-competent host species, including house finch and house sparrow, were common bloodmeal sources for both vector species across several biomes and could account for WNV maintenance and amplification in these areas. Highly competent American crow, western scrub-jay and yellow-billed magpie also were fed upon often when available and are likely important as amplifying hosts for WNV in some areas. Neither species fed frequently on humans (Cx. pipiens complex [0.4%], Cx. tarsalis [0.2%]), but with high abundance, both species could serve as both enzootic and bridge vectors for WNV.     

Host-feeding habits of Culex and other mosquitoes (Diptera: Culicidae) in the Borough of Queens in New York City,with characters and techniques for identification of Culex mosquitoes

The host-feeding patterns of mosquitoes (n = 247) collected in the Borough of Queens in New York City in July and August 2000 were investigated using an indirect ELISA and a polymerase chain reaction (PCR)-heteroduplex assay. Culex pipiens L. and Cx. restuans Theobald fed primarily on birds, and their feeding habits support their implication as enzootic vectors of West Nile virus. Culex salinarius Coquillett and Coquillettidia perturbans (Walker) fed mainly on mammals, with fewer blood meals taken from birds, and these two species are potential bridge vectors of West Nile virus. Culex mosquitoes took blood meals (n = 54) from 11 different avian species. Only the northern cardinal (Cardinalis cardinalis), American robin (Turdus migratorius), and Brown-headed cow bird (MolIothrus ater) were fed upon by all three Culex species. Multiple blood feedings on avian hosts were detected in Cx. pipiens and Cx. restuans. Species identifications of Culex mosquitoes made using morphological characteristics were confirmed with a PCR assay that employed species-specific primers. All Cx. pipiens (n = 20) and Cx. salinarius (n = 10) specimens were correctly identified, but three (20%) of 15 Cx. restuans were misidentified as Cx. pipiens.     

Evaluation of potential West Nile virus vectors in Volgograd region, Russia, 2003 (Diptera: Culicidae): species composition, bloodmeal host utilization, and virus infection rates of mosquitoes

Potential West Nile virus (family Flaviviridae, genus Flavivirus, WNV) vectors were assessed during 2003 at indoor and outdoor collection sites in urban Volgograd, Russia, and in three nearby towns and surrounding rural areas. In total, 9,182 female mosquitoes comprising 13 species in six genera were collected. Relative abundance and bloodmeal host utilization differed temporarily and spatially. During June and July in Volgograd, Aedes vexans (Meigen) (85.4%) and Culex p. pipiens L. (7.6%) were the two most abundant species collected indoors, whereas during August, Cx. p. pipiens was the dominant species, accounting for 87.9% of specimens collected. Two WNV-positive mosquito pools were detected in August: one pool was composed of Cx. p. pipiens and the other pool of Culex modestus Ficalbi. Anopheles messeae Falleroni, Aedes caspius (Pallas), Ae. vexans, Cx. modestus, and Cx. p. pipiens used both humans and birds as bloodmeal sources. In urban areas, 20.4% of the Cx. p. pipiens fed on humans, 58.1% fed on chickens, and six specimens were positive for both chicken and human blood. Culex p. pipiens collected from flooded basements were predominantly autogenous (91.7%), whereas adult females resting in buildings with dry basements were composed of 67.5% anautogenous and 32.5% autogenous specimens. Our data suggest that the primary WNV vectors in the Volgograd region were Cx. p. pipiens and Cx. modestus and that intense transmission of WNV to humans in urban areas during the epidemic of 1999 may have been facilitated by the abundance and concentration of anautogenous Cx. p. pipiens in multistory buildings. The role of autogenous Cx. p. pipiens in urban transmission remains unresolved.     

Host-feeding patterns of Culex pipiens and other potential mosquito vectors (Diptera: Culicidae) of West Nile virus (Flaviviridae) collected in Portugal

The host blood-feeding patterns of mosquito vectors affects the likelihood of human exposure to zoonotic pathogens, including West Nile Virus (family Flaviviridae, genus Flavivirus, WNV). In Portugal, data are unavailable regarding the blood-feeding habits of common mosquito species, including Culex pipiens L., considered the primary vector of WNV to humans. The sources of bloodmeals in 203 blood-fed mosquitoes of nine species collected from June 2007 to November 2010 in 34 Portuguese counties were analyzed by sequencing cytochrome-b partial fragments. Cx. pipiens was the most common species collected and successfully analyzed (n = 135/78). In addition, blood-fed females of the following species were analyzed: Ochlerotatus caspius Pallas (n = 20), Culex theileri Theobald (n = 16), Anopheles maculipennis s.l. Meigen (n = 10), Culiseta longiareolata Macquart (n = 7), Aedes aegypti L. (n = 6), Culex perexiguus Theobald (n = 3), Culiseta annulata Schrank (n = 3), and Ochlerotatus detritus Haliday (n = 3). The Cx. pipiens mosquitoes fed predominantly on birds (n = 55/78, 70.5%), with a high diversity of avian species used as hosts, although human blood was identified in 18 specimens (18/78, 23.1%). No significant differences were found between the host-feeding patterns of blood-fed Cx. pipiens collected in residential and nonresidential habitats. The occurrence of human derived blood meals and the presence of a mix avian-human bloodmeal accordingly suggest this species as a potential vector of WNV. Therefore, in Portugal, Cx. pipiens may play a role both in the avian-to-avian enzootic WNV cycle and in the avian-to-mammal transmission. In this context, the identity of Cx. pipiens (considering the forms molestus and pipiens) and the potential consequence on feeding behavior and WNV transmission are discussed.     

Bird-baited traps for surveillance of West Nile mosquito vectors: effect of bird species, trap height, and mosquito escape rates

Host-seeking mosquitoes were sampled in bird-baited traps at four sites in New York state in 2003-2004. Trap placement and efficacy of chickens, Gallus gallus domesticus L., as bait compared with house sparrows, Passer domesticus L., an important reservoir of West Nile virus (family Flaviviridae, genus Flavivirus, WNV), was evaluated. Each site had a chicken-baited trap near ground level (approximately 1.5 m) and in the tree canopy (approximately 9 m), and a house sparrow-baited trap at ground level and canopy level. Each trap allowed mosquito access to birds on one end, and an inner mesh screen blocked bird access on the other end. The two most abundant mosquitoes, Culex restuans Theobald and Culex pipiens pipiens L., were differentiated using molecular characters. In 2003, Cx. restuans and Cx. p. pipiens made up 88% of total mosquito catch. In 2004, Cx. restuans comprised 43% of total catch and Cx. p. pipiens comprised 33%. The remaining species representing at least 1% of total catch were Ochlerotatus trivittatus (Coquillett), Coquilletidia perturbans (Walker), and Culiseta morsitans (Theobald). Capture rates were similar for chicken and house sparrow-baited traps; however, significantly more mosquitoes were captured in the canopy for both bird species. Cx. restuans preferred canopy traps, whereas equal numbers of Cx. p. pipiens were captured at ground and canopy levels. Mosquitoes were more likely to escape (74%) when excluded from birds than when allowed free access to birds (54%). Sentinel bird surveillance for WNV can be improved by trapping in the tree canopy in addition to ground level to capture the most important avian vectors.     

Impact of aerial spraying of pyrethrin insecticide on Culex pipiens and Culex tarsalis (Diptera: Culicidae) abundance and West Nile virus infection rates in an urban/suburban area of Sacramento County, California

In response to an epidemic amplification of West Nile virus (family Flaviviridae, genus Flavivirus, WNV), the Sacramento and Yolo Mosquito and Vector Control District (SYMVCD) sprayed ultralow-volume (ULV) formulations of pyrethrin insecticide (Evergreen EC 60-6: 6% pyrethrin insecticide, 60% piperonylbutoxide; MGK, Minneapolis, MN, applied as 0.003 kg/ha [0.0025 lb/acre] ) over 218 km2 in north Sacramento and 243.5 km2 in south Sacramento on three consecutive evenings in August 2005. We evaluated the impact of this intervention in north Sacramento on the abundance and WNV infection rates of Culex pipiens L. and Culex tarsalis Coquillett. Mortality rates of caged Cx. tarsalis sentinels ranged from 0% under dense canopy to 100% in open fields. A comparison of weekly geometric mean mosquito abundance in CO2-baited traps in sprayed and unsprayed areas before and after treatment indicated a 75.0 and 48.7% reduction in the abundance of Cx. pipiens and Cx. tarsalis, respectively. This reduction was statistically significant for Cx. pipiens, the primary vector of WNV, with highest abundance in this urban area, but not for Cx. tarsalis, which is more associated with rural areas. The infection rates of WNV in Cx. pipiens and Cx. tarsalis collected from the spray zone were 8.2 and 4.3 per 1,000 female mosquitoes in the 2 wk before and the 2 wk after applications of insecticide, respectively. In comparison, WNV infection rates in Cx. pipiens and Cx. tarsalis collected at same time interval in the unsprayed zone were 2.0 and 8.7 per 1,000, respectively. Based on the reduction in vector abundance and its effects on number of infective bites received by human population, we concluded that the aerial application ofpyrethrin insecticide reduced the transmission intensity of WNV and decreased the risk of human infection.     

Experimental studies on comparison of the potential vector competence of four species of Culex mosquitoes in China to transmit West Nile virus

To assess the risk that indigenous mosquitoes in China are capable of transmitting and sustaining West Nile virus (WNV), four important Culex mosquito species, Culex tritaeniorhynchus, Culex modestus, Culex pipiens pallens, and Culex pipiens quinquefasciatus, were allowed to feed on the artificial infectious blood meal with WNV dose of 10(6.8) plaque-forming unit/ml and tested approximately 2 wk later to determine infection and transmission rates. The results indicated that four Culex mosquitoes were competent laboratory vectors of WNV. The infection rates and transmission rates were statistical differences among different species of mosquito (chi2 = 20.620, P = 0.000; chi2 = 15.020, P = 0.005, respectively). The highest infection rate and transmission rate were obtained with Cx. tritaeniorhynchus (87.5 and 74.2%, respectively).     

West Nile virus from female and male mosquitoes (Diptera: Culicidae) in subterranean, ground, and canopy habitats in Connecticut

In total, 93,532 female mosquitoes (Diptera: Culicidae) were captured in traps placed in subterranean (catch basin), ground (approximately 1.5 m above ground), and canopy (approximately 7.0 m above ground) habitats in Stamford and Stratford, CT, during 2003-2005. Culex pipiens L. was the most abundant (64.8%) of the 31 species identified. Significantly greater numbers of Cx. pipiens were captured in canopy-placed mosquito magnet experimental traps, and significantly greater numbers were collected in catch basin-placed (Centers for Disease Control) CDC traps than in CDC traps placed elsewhere. Culex restuans Theobald was captured in significantly greater numbers in traps placed in catch basins. Aedes vexans (Meigen), Aedes cinereus Meigen, and Aedes cantator (Coquillett) were significantly more abundant in ground traps. In total, 429 isolations of West Nile virus (WNV) were made from seven species of mosquitoes from late June through the end of October during 2003 through 2005. Three hundred ninety-eight (92.8%) isolates were from Cx. pipiens. Others were from Cx. restuans (n = 16), Culex salinarius Coquillett (n = 5), Ae. vexans (n = 4), Ae. cantator (n = 3), Aedes triseriatus (Say) (n = 2), and Ae. cinereus (n = 1). Multiple isolates from Cx. pipiens were made each week, primarily during the later part of July through the end of September. Weekly minimum infection rates (MIRs) were lower in 2004 (highest weekly MIR = 7.1) when no human cases were reported in Connecticut in comparison with 2003 and 2005 (highest weekly MIR = 83.9) when human cases were documented. Frequencies of infected pools were significantly higher in Cx. pipiens captured in traps in the canopy and significantly higher in catch basin placed traps than in traps at ground level. The physiological age structure of Cx. pipiens captured in the canopy was significantly different from that of Cx. pipiens collected in catch basins. Invariably, Cx. pipiens captured in the canopy were nulliparous or parous with ovaries in Christophers' stage 2, whereas 58.7% of the females captured in catch basins possessed ovaries filled with mature oocytes in Christophers' stage 5. Our results suggest that females in the canopy are seeking hosts, and after digestion of the bloodmeal and development of mature oocytes, they descend to catch basins for shelter and deposition of eggs. WNV was isolated from three, one, and two pools of male Cx. pipiens captured in catch basin-, ground-, and canopy-placed traps, respectively, and from six nulliparous Cx. pipiens females collected in the canopy. Weekly MIR ranged from 1.2 to 31.1 per 1,000 male specimens. These data show that mosquitoes become infected by means other than by blood feeding, possibly by transovarial transmission. The placement of traps in tree canopies and in catch basins can be used to augment current practices of placement of traps near the ground for surveillance of mosquitoes infected with WNV and for studies of the ecology of WNV.     

West Nile virus-infected dead corvids increase the risk of infection in Culex mosquitoes (Diptera: Culicidae) in domestic landscapes

A comparative study of West Nile virus (family Flaviviridae, genus Flavivirus, WNV) infection rates in Culex mosquitoes collected at 13 sites, seven reporting WNV-positive dead corvids (case sites) and six without reported dead birds (control sites) was conducted in Davis, CA, from 14 to 21 July at the beginning of the 2006 WNV outbreak. In total, 3051 Culex mosquitoes were collected using gravid traps and CO2-baited traps; WNV-infected mosquitoes were only collected with CO2-baited traps. WNV-infected Culex pipiens L. were collected at one of the seven case sites. Six of seven case sites yielded WNV-infected Culex tarsalis Coquillett, whereas only one of six control sites had WNV-infected Cx. tarsalis. Overall, the odds of finding WNV-positive mosquitoes were 19.75 times greater at sites reporting a WNV-infected dead corvid than sites without a WNV-infected dead corvid. Maximum likelihood estimates of the overall infection rates at the case sites were 3.48/1000 for Cx. tarsalis and 8.69/1000 for Cx. pipiens compared with 1.02/1000 in Cx. tarsalis collected at the control sites. Results indicate that Cx. tarsalis was important in early season enzootic transmission within Davis and that sites reporting WNV-infected dead corvids are areas to focus control and surveillance efforts.     

Horse-, bird-, and human-seeking behavior and seasonal abundance of mosquitoes in a West Nile virus focus of southern France

After 35 yr of disease absence, West Nile virus (family Flaviviridae, genus Flavivirus, WNV) circulation has been regularly detected in the Camargue region (southern France) since 2000. WNV was isolated from Culex modestus Ficalbi, which was considered the main vector in southern France after horse outbreaks in the 1960s. Recent WNV transmissions outside of the Cx. modestus distribution suggested the existence of other vectors. To study potential WNV vectors, horse- and bird-baited traps and human landing collections of mosquitoes were carried out weekly from May to October 2004 at two Camargue sites: one site in a wet area and the other site in a dry area, both chosen for their past history of WNV transmission. At the wet site, the most abundant species in bird-baited traps were Culex pipiens L. and Cx. modestus; both species also were found in lower proportions on horses and humans. The most abundant species in horse-baited traps and human landing collections were Aedes caspius (Pallas), Aedes vexans (Meigen), and Anopheles hyrcanus (Pallas) sensu lato; some of these species were occasionally collected with avian blood at the end of the summer. Anopheles maculipennis Meigen sensu lato was an abundant horse feeder, but it was rarely collected landing on human bait and never contained avian blood. At the dry site, Cx. pipiens was the most abundant species in bird- and horse-baited traps. The seasonal and circadian dynamics of these species are analyzed, and their potential in WNV transmission in Camargue discussed.     

West Nile virus investigations in South Moravia, Czechland

Seven virus isolates were obtained from 11,334 mosquitoes after the 1997 Morava River flooding in South Moravia (Czech Republic): 6 strains of Tahyna bunyavirus, California antigenic group (5 from Aedes vexans, 1 from Ae. cinereus), and 1 strain of West Nile flavivirus (WNV) from Culex pipiens. In 1999, one isolate of Tahyna virus from Ae. vexans and one isolate of WNV from Cx. pipiens were recovered from a total of 14,354 mosquitoes examined in the same area, whereas no virus was detected there in 1,179 overwintering mosquitoes (mostly Cx. pipiens) in March 2000. The infection rate of mosquitoes with arboviruses was significantly higher in 1997, the year of the flood and an enormously high population density of mosquitoes. Antibodies neutralizing WNV were detected in 13 of 619 (2.1%) hospitalized patients or persons seeking outpatient clinics of the area in 1997. Five of the seroreactors revealed clinical symptoms compatible with West Nile fever: in 2 of them (children), recent infection with WNV was confirmed by a significant increase of antibody titer between acute and convalescent serum samples.     

Impact of trap elevation on estimates of abundance, parity rates, and body size of Culex pipiens and Culex restuans (Diptera: Culicidae)

Trapping success, abundance, parity rate, and body size indices of Culex pipiens (L.) and Culex restuans Theobald, important vectors of West Nile virus (family Flaviviridae, genus Flavivirus, WNV), were determined for specimens captured from both ground level (approximately 1-m) and elevated (approximately 6- to 7-m) CO2-baited CDC miniature light traps. Mosquitoes were collected from six study sites in Albany County, New York, from mid-May to mid-October 2004. There was no significant difference in abundance or mean parity for either Cx. pipiens or Cx. restuans between the elevated and the ground-level traps. Mosquitoes collected in elevated traps tended to be larger than mosquitoes collected at ground level. Elevated traps captured significantly fewer mosquitoes per trap-night than did ground traps, but a greater proportion of those captured were Culex spp. Therefore, elevated traps more efficiently captured Cx. pipiens and Cx. restuans with similar reproductive status and body size and reduced the time necessary to sort and identify collections. These findings may be beneficial to researchers or surveillance programs focusing on Cx. pipiens or Cx. restuans.     

West Nile virus infection in mosquitoes in the mid-south USA, 2002-2005

West Nile virus (family Flaviviridae, genus Flavivirus, WNV) was first detected in the Tennessee Valley and in Alabama in August 2001. In summer 2002, intensive viral activity was seen, but in subsequent years, viral activity settled into an enzootic pattern. Here, we report an analysis of viral activity in the mosquito fauna in the Mid-South from 2002 (the first year viral activity was detected in mosquitoes) through 2005. Eight mosquito species were infected with WNV during 2002. However, viral activity was only detected in four species--Culex salinarius Coquillett, Culex erraticus Dyar & Knab, Coquillettidia perturbans Walker, and Aedes vexans Meigen--in multiple years. The greatest number of positive pools was in Cx. erraticus and Cx. salinarius. Despite being specifically targeted for collection, Aedes albopictus Skuse was only found to be infected during the epiornitic year (2002), suggesting that under enzootic transmission conditions its role as a bridge vector in the region may not be significant. Virus-positive pools of Cx. erraticus were identified from winter-resting and early season dry ice-baited trap collections in 2005, implicating this species in WNV overwintering in Alabama. Molecular analysis of individuals initially identified as members of the Culex pipiens L. complex suggested that alleles characteristic of Cx. pipiens predominated in mosquitoes collected in Huntsville, AL, whereas alleles in the Auburn, AL, population were predominately characteristic of Culex quinquefaciatus Say. The southern boundary of the overlap zone of the two species seems to be located primarily between Huntsville and Auburn, a distance of 350 km.     

Role of corvids in epidemiology of west Nile virus in southern California

The invasion of different southern California landscapes by West Nile virus (WNV) and its subsequent amplification to epidemic levels during 2004 enabled us to study the impact of differing corvid populations in three biomes: the hot Colorado desert with few corvids (Coachella Valley), the southern San Joaquin Valley (Kern County) with large western scrub-jay but small American crow populations, and the cool maritime coast (Los Angeles) with a large clustered American crow population. Similar surveillance programs in all three areas monitored infection rates in mosquitoes, seroconversion rates in sentinel chickens, seroprevalence in wild birds, numbers of dead birds reported by the public, and the occurrence of human cases. Infection rates in Culex tarsalis Coquillett and sentinel chicken seroconversion rates were statistically similar among all three areas, indicating that highly competent mosquito hosts were capable of maintaining enzootic WNV transmission among less competent and widely distributed avian hosts, most likely house sparrows and house finches. In contrast, infection rates in Culex pipiens quinquefasciatus Say were statistically higher in Kern and Los Angeles counties with elevated corvid populations than in Coachella Valley with few corvids. Spatial analyses of dead corvids showed significant clusters near known American crow roosts in Los Angeles that were congruent with clusters of human cases. In this area, the incidence of human and Cx. p. quinquefasciatus infection was significantly greater within corvid clusters than without, indicating their importance in virus amplification and as a risk factor for human infection. In contrast the uniform dispersion by territorial western scrub-jays resulted in a high, but evenly distributed, incidence of human disease in Kern County.     

Oviposition activity patterns and West Nile virus infection rates for members of the Culex pipiens complex at different habitat types within the hybrid zone, Shelby County, TN, 2002 (Diptera: Culicidae)

Oviposition activity and West Nile virus (family Flaviviridae, genus Flavivirus, WNV) infection rates were assessed for members of the Culex pipiens complex from July through December 2002 by using gravid traps placed at four ecologically different sites in the southern portion of the hybrid zone in Shelby County, TN. Molecular assays identified three members of the Cx. pipiens complex: Cx. pipiens pipiens L., Cx. p. quinquefasciatus Say, and Cx. p. pipiens-Cx. p. quinquefasciatus hybrids (hybrids). The Cx. pipiens complex accounted for 90% of mosquitoes collected in gravid traps. All 285 WNV-positive mosquitoes were Culex mosquitoes, and 277 (97%) were Cx. pipiens complex mosquitoes. Infection rates among members of the Cx. pipiens complex were not significantly different. Infection rates were significantly higher at two urban sites than at a rural site, and WNV was not detected at a forested site. At urban sites, abundances of members of the Cx. pipiens complex corresponded to a simple latitude model of the hybrid zone. Cx. p. quinquefasciatus was most abundant (46.4%), followed by hybrids (34.1%) and Cx. p. pipiens (19.5%). The relative abundances at a rural site were reversed with Cx. p. pipiens (48.4%) being most abundant. This demonstrates that spatial habitat variation may profoundly influence the distribution of members of the Cx. pipiens complex within the hybrid zone. Members of the Cx. pipiens complex did not display different oviposition patterns. However, oviposition patterns assessed hourly at urban and rural sites were significantly different. At urban sites, oviposition activity of Cx. pipiens complex mosquitoes was bimodal with an evening peak associated with sunset and a morning peak associated with sunrise. At the rural site, the evening peak was pronounced and the morning peak weak and similar to nighttime activity.     

Considerations for accurate identification of adult Culex restuans (Diptera: Culicidae) in field studies

Understanding the ecology and behavior of different mosquito species (Diptera: Culicidae) is essential for identifying their role in disease transmission cycles and public health risk. Two species of Culex mosquitoes in the northeastern United States, Culex pipiens L. and Culex restuans Theobald, have been implicated in enzootic transmission of West Nile virus (family Flaviviridae, genus Flavivirus, WNV). Despite the difficulty of differentiating these two species as adults, many public health workers and vector biologists collecting adults in the field separate these species based on external morphology. This approach is often used rather than examination of dissected male genitalia or polymerase chain reaction (PCR)-based diagnostics due to time or cost constraints. We evaluated the reliability of seven published morphological characters to differentiate adults of these species by comparing blindly scored morphology with PCR-based confirmations. Our study demonstrates that morphological identification of Cx. pipiens is marginal and often not reliable for Cx. restuans. We also examined error rates with molecular-based approaches. DNA samples were contaminated with as little as one leg from another species. We conclude that to fully understand the respective roles of Culex species in the epidemiology of WNV and other pathogens, more attention should be paid to these considerations for accurate species identification.     

Avian host and mosquito (Diptera: Culicidae) vector competence determine the efficiency of West Nile and St. Louis encephalitis virus transmission

The ability of the invading NY99 strain of West Nile virus (WNV) to elicit an elevated viremia response in California passerine birds was critical for the effective infection of Culex mosquitoes. Of the bird species tested, Western scrub jays, Aphelocoma coerulescens, produced the highest viremia response, followed by house finches, Carpodacus mexicanus, and house sparrows, Passer domesticus. Most likely, few mourning, Zenaidura macroura, or common ground, Columbina passerine, doves and no California quail, Callipepla californica, or chickens would infect blood-feeding Culex mosquitoes. All Western scrub jays and most house finches succumbed to infection. All avian hosts produced a lower viremia response and survived after infection with an endemic strain of St. Louis encephalitis virus. Culex species varied in their susceptibility to infection with both viruses, with Culex stigmatosoma Dyar generally most susceptible, followed by Culex tarsalis Coquillett, and then Culex p. quinquefasciatus Say. Populations within Culex species varied markedly in their susceptibility, perhaps contributing to the focality of WNV amplification. Transmitting female Cx. tarsalis expectorated from six to 3,777 plaque-forming units (PFU) of WNV during transmission trials, thereby exposing avian hosts to a wide range of infectious doses. Highly susceptible house finches and moderately susceptible mourning doves were infected by subcutaneous inoculation with decreasing concentrations of WNV ranging from 15,800 to <0.3 PFU. All birds became infected and produced comparable peak viremias on days 2-3 postinoculation; however, the rise in viremia titer and onset of the acute phase of infection occurred earliest in birds inoculated with the highest doses. WNV virulence in birds seemed critical in establishing elevated viremias necessary to efficiently infect blood feeding Culex mosquitoes.     

Culex restuans (Diptera: Culicidae) relative abundance and vector competence for West Nile Virus

The abundance and vector competence of Culex restuans Theobald and Culex pipiens L. were compared to determine the relative importance of these species as West Nile virus (WNV) vectors in the northeastern United States. Abundance was estimated from egg raft collections at 12 sites in Albany, Suffolk, and Richmond counties, New York, during July, August, and September 2002 and 2003. Cx. restuans was more abundant than Cx. pipiens in both urban and rural areas, comprising 86% of 1,623 egg rafts collected. Vector competence for WNV was estimated after feeding on an artificial bloodmeal and in vitro transmission assays. The vector competence of the two species for WNV was similar, but the dynamics of infection seems to be mosquito species dependent. These findings suggest an important role for Cx. restuans in WNV transmission cycles in New York.