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.     

European experience with the West Nile virus ecology and epidemiology: could it be relevant for the New World?

A review of West Nile virus (WNV) and the epidemiology of West Nile fever (WNF) in Europe is presented. European epidemics of WNF reveal some general features. They usually burst out with full strength in the first year, but few cases are observed in the consecutive 1 to 2 (exceptionally 3) years, whereas smaller epidemics or clusters of cases only last for one season. The outbreaks are associated with high populations of mosquitoes (especially Culex spp.) caused by flooding and subsequent dry and warm weather, or formation of suitable larval breeding habitats. Urban WNF outbreaks associated with Culex pipiens biotype molestus are dangerous. Natural (exoanthropic, sylvatic) foci of WNV characterized by the wild bird-ornithophilic mosquito cycle probably occur in many wetlands of climatically warm and some temperate parts of Europe; these foci remain silent but could activate under circumstances supporting an enhanced virus circulation due to appropriate abiotic (weather) and biotic (increased populations of vector mosquitoes and susceptible avian hosts) factors. It is very probable that WNV strains are transported between sub-Saharan Africa and Europe by migratory birds. The surveillance system for WNF should consist of four main components: (1) monitoring of mosquito populations and their infection rate; (2) wild vertebrate surveys; (3) sentinel birds (domestic ducks rather than chickens); and (4) monitoring of human disease. In the case of persisting high risk of WNF for humans and equids in certain enzootic areas, immunization against WNF should be considered. For that purpose a commercially available, cross-protective vaccine against Japanese encephalitis could be used.     

Estimation du risque d’introduction du virus de la fièvre de la vallée du Rift en Tunisie par le moustique Culex pipiens

The mosquito Culex pipiens has been involved as vector of the West Nile virus in Tunisia. Its bio-ecological characteristics in combination with some environmental factors have favoured the emergence of this virus in a West-Nile free zone. This leads to question about the potential risk of introducing another arbovirus, the Rift Valley fever (RVF) virus, in Tunisia from neighbouring countries where RVF circulates. In this study, we have evaluated the vector competence of different populations of Cx. pipiens towards two strains of RVF virus, the virulent ZH548 and the avirulent Clone 13 by experimental infections and the genetic differentiation of these populations of Cx. pipiens using four microsatellite loci. We found disseminated infection rates ranging from 0% to 14.7% and a high genetic differentiation among populations without any geographical pattern (no isolation by distance). Thus, although Cx. pipiens is able to sustain an amplification of RVF virus, viral dissemination through mosquito dispersal would be unlikely. However, as RVF is an emerging disease transmitted by several other potential mosquito species (e.g. Ochlerotatus caspius), attention should be maintained to survey livestock and mosquitoes in Tunisia.     

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).     

Culex pipiens (Diptera: Culicidae): a bridge vector of West Nile virus to humans

Host-feeding patterns of Culex pipiens L. collected in southwest suburban Chicago in 2005 were studied using polymerase chain reaction (PCR) and DNA sequencing techniques. Culex spp. mosquitoes, most identified to Cx. pipiens and the remainder to Cx. restuans by PCR, had fed on 18 avian species, most commonly American robin (Turdus migratorious), house sparrow (Passer domesticus), and mourning dove (Zenaida macroura). Additional blood meals were derived from four mammal species, primarily humans and raccoons (Procyon lotor). During a West Nile virus (WNV) epidemic in 2005, West Nile virus (WNV) RNA was detected in heads and thoraces of five Cx. pipiens (n = 335, 1.5%) using quantitative PCR. The hosts of these virus-infected, blood-fed mosquitoes included two American robins, one house sparrow, and one human. This is the first report of a WNV-infected Cx. pipiens mosquito collected during an epidemic of WNV that was found to have bitten a human. These results fulfill a criterion for incrimination of Cx. pipiens as a bridge vector.     

Host-feeding patterns of native Culex pipiens and invasive Aedes albopictus mosquitoes (Diptera: Culicidae) in urban zones from Barcelona, Spain

The feeding patterns of haematophagous arthropods are of major importance in the amplification and transmission of infectious disease agents to vertebrate hosts, including humans. The establishment of new vector populations in nonnative range might alter transmission networks. The Asian tiger mosquito Aedes albopictus (Skuse) represents an example of how an invasive species can alter the risk of viral transmission to humans. Blood meal molecular identification from two sympatric mosquito species (the invasive Ae. albopictus and the native Culex pipiens) was carried out by polymerase chain reaction-based methods. Samples were collected in Barcelona metropolitan area, Spain, from June to October 2009 as part of a monitoring-control program. Blood meals were identified to the species level in 30 Ae. albopictus and 43 Cx. pipiens. Ae. albopictus acquired blood exclusively from human hosts (100%), whereas Cx. pipiens fed on a diversity of avian and mammalian hosts, including 35.7% of blood meals from humans. Based on mosquito diet, our results suggest that the Ae. albopictus invasion in Spain might increase the risk of virus transmission to humans and could support local outbreaks of imported tropical viruses such as dengue and chikungunya. However, in the studied area, the presence of this invasive species would have a negligible effect on the transmission of zoonotic agents such as West Nile virus. However, Cx. pipiens could amplify and transmit West Nile virus, but avian contribution to its diet was lower than that reported in North America. Feeding patterns of these mosquito species may help to understand the flavivirus outbreaks recently reported in southwestern Europe.     

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.     

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.     

Identification and characterization of single nucleotide polymorphisms (SNPs) in Culex theileri (Diptera: Culicidae)

Culex theileri Theobald (Diptera: Culicidae) is one of the most common mosquito species in northeastern Turkey and serves as a vector for various zoonotic diseases including West Nile virus. Although there have been some studies on the ecology of Cx. theileri, very little genetic data has been made available. We successfully sequenced 11 gene fragments from Cx. theileri specimens collected from the northeastern part of Turkey. On average, we found a Single nucleotide polymorphism every 45 bp. Transitions outnumbered transversions, at a ratio of 2:1. This is the first report of genetic polymorphisms in Cx. theileri and Single nucleotide polymorphism discovered from this study can be used to investigate population structure and gene-environmental interactions.     

Hematophagous Diptera collected from a horse and paired carbon dioxide-baited suction trap in southern California: relevance to West Nile virus epizootiology

Hematophagous Diptera landing on a horse were removed by vacuum, and their numbers were related to a paired carbon dioxide-baited suction trap at three locations in southern California where West Nile virus activity was high during the preceding year. Insects collected from the horse included mosquitoes (nine species), biting midges (Culicoides sonorensis Wirth & Jones), and black flies (Simulium bivittatum Malloch). Mosquitoes were predominantly collected from the head, crest, withers, neck, chest, and shoulders of the horse, whereas biting midges and black flies were predominantly collected from the ventral midline of the horse. Culex erythrothorax Dyar was by far the most abundant mosquito species collected overall. Frequency of engorgement for mosquitoes captured from the horse ranged by species from zero to 58.3%, with Culex pipiens quinquefasciatus Say having the lowest value (16.7% or one of six mosquitoes) of species that fed on the horse. The number of insects captured at the horse and paired CO2-baited suction trap was not different for Anopheles franciscanus McCracken, Culex tarsalis Coquillett, and S. bivittatum. Cx. p. quinquefasciatus was captured in greater numbers in the CO2-baited suction trap, whereas Anopheles hermsi Barr & Guptavanji, Cx. erythrothorax, Culiseta inornata (Williston), and Culiseta particeps (Adams) were captured in greater numbers from the horse. The horse biting rate was very low for Cx. p. quinquefasciatus, intermediate for Cx. tarsalis, and very high for Cx. erythrothorax. Both Cx. tarsalis and Cx. erythrothorax should be considered likely epizootic vectors of West Nile virus to horses in rural southern California.     

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.     

Infectivity and pathogenicity of a novel baculovirus, CuniNPV from Culex nigripalpus (Diptera: Culicidae) for thirteen species and four genera of mosquitoes

The infectivity and pathogenicity of newly discovered baculovirus, CuniNPV (family Baculoviridae, genus Nucleopolyhedrovirus) originally isolated from the mosquito Culex nigripalpus Theobald, was evaluated in laboratory bioassys against thirteen species and four genera of mosquitoes native to the northeastern U.S. Purified virus at a dosage rate of 1.6 x 10(7) occlusion bodies/ml with 10 mM Mg2+ added was used in exposures with second through fourth instars at temperatures ranging from 17 to 27 degrees C. High infection rates and accompanying mortality were achieved in Cx. pipiens L. (83.0-14.4%), Cx. pipiens f. molestus (80.4% infection), and Cx. salinarius Coquillett (48.0-43.1%). Cx. restuans Theobald was also susceptible but infection rates were lower (21.3-12.5%). The gross pathology associated with infection was identical to that reported in Cx. nigripalpus. Infected larvae were lethargic and were often suspended at the water surface. Development of CuniNPV was observed in the nuclei of the midgut epitheial cells in the gastric caeca and posterior region of the stomach of host larvae. One hundred percent mortality was observed in all larvae that exhibited gross symptoms of infection within 4-d p.i. Cx. territans Walker (subgenus Neoculex Dyar) was the only Culex mosquito that was not susceptible. No infections were obtained with any species of Aedes [Ae. vexans (Meigen)], Culiseta [Culiseta morsitans (Theobald)] or Ochlerotatus [Ochlerotatus canadensis (Theobald), Oc. cantator (Coquillett), Oc. communis (De Geer), Oc. excrucians (Walker), Oc. japonicus (Theobald), Ochlerotatus stimulans (Walker), and Ochlerotatus triseriatus (Coquillett)]. The host range of CuniNPV appears to be restricted to Culex mosquitoes within the subgenus Culex. An inhibitory effect on transmission of CuniNPV was observed when a liver powder/Brewer's yeast mixture was used as a source of food reinforcing the critical role of Mg2+ and sensitivity of the infection process to the presence other divalent cations (Cu2+, Fe2+, and Zn2+) in the larval medium that interfered with the infection process. The high infectivity and pathogenicity of CuniNPV for the principal vectors of West Nile virus in North America make CuniNPV an attractive candidate for future development as a biopesticide.     

Host feeding of mosquitoes (Diptera: Culicidae) associated with the recurrence of Rift Valley fever in Egypt

In 1993, Rift Valley fever (RVF) virus reappeared in Egypt. We determined the prevalence and feeding patterns of mosquitoes in 5 villages where the virus was active. Of 10 species recovered, Aedes caspius (Pallas), Culex pipiens L., Cx. antennatus (Becker), and Cx. perexiguus Theobald constituted 99% of > 35,000 mosquitoes captured in dry ice-baited CDC light traps. Ae. caspius was most prevalent, except at Nag' El Hagar where it was replaced by Cx. perexiguus. Cx. pipiens ranked 2nd, except at Nag' El Ghuneimiya, where it was replaced by Cx. antennatus. Most blood meals analyzed by an enzyme-linked immunosorbent assay reacted to > or = 1 antiserum. Cx. pipiens was mainly anthropophagic, and therefore may have been the main vector of RVF virus among humans. Ae. caspius feeds were chiefly from humans, bovines, and equines. Cx. antennatus and Cx. perexiguus fed generally on bovines. Mixed blood meals from humans and RVF virus susceptible animals were identified in the predominant mosquitoes. Prevalence and host selection, as well as predicted probability for a blood meal being interrupted, indicated that Ae. caspius may have served as a bridge vector between humans and bovines in 4 of the villages. Cx. perexiguus may have played this role at Nag' El Hagar. Because potential vectors are abundant, susceptible domestic animals are associated closely with humans, and surveillance of imported livestock is not systematic, we conclude that RVF virus sporadically will recur in Egypt.     

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.     

云南景洪市蚊虫季节消长与乙型脑炎流行关系研究

为掌握云南景洪市蚊虫季节消长特点及其与乙型脑炎流行的关系,为该疾病防控提供有效依据,2007年10月至2008年9月本研究在云南景洪市普文镇畜圈和人房每月采用诱蚊灯方法通宵捕蚊,对采获的蚊虫使用白纹伊蚊C6/36细胞培养法分离乙脑病毒,并对分离到的病毒进行分子生物学鉴定.收集2007～2008年景洪市乙脑疫情资料进行乙...     

The influence of weather conditions on the epidemiology of vector-borne diseases by the example of West Nile fever in Russia

Climate changes must influence the incidence of vector-borne infections, but their effects cannot be revealed due to lack of long-term observations. The impact of short-term weather changes may be used as a model. In Russia the biggest numbers of clinical cases of mosquito-borne West Nile infection were registered in 1999 in Volgograd and Astrakhan regions. The analysis of climatic dataset since 1900 shows that 1999 was the hottest year in Volgograd in the 20th century due to a very mild winter (December-March) and a rather hot summer (June-September). The author of the article puts forward a hypothesis that high winter temperatures favored the survival of over-wintering mosquito vectors, and high summer temperature facilitated the growth of the virus in the mosquitoes, as well as propagation of the mosquitoes themselves. The author assumes that conventional threshold temperatures for "beneficial for WNF conditions" in Russia are > or = 3 degrees C in winter, and > or = 22 degrees C in summer. These conditions coincided only in 1948 and 1999. In Astrakhan the "beneficial for WNF conditions" were registered in 30 out of 147 years of observation, and in 12 years between 1964 and 2003. This is not surprising that Astrakhan region is endemic for WNF in accordance with clinical and epidemiological data collected since the sixties. These findings give some hints on the WNF predisposing factors, as well as possibility of weather surveillance and prediction of WNF outbreaks in temperate climatic zones such as Southern Russia.     

Specific antibodies in convalescents after West Nile fever

Blood serum samples were obtained from patients with West Nile fever (WNF) from the city of Volgograd and its region within the disease weeks 1-3 (July-October 1999), month 6-7 (May_June 2000) and month 12-18 (May-June 2001) after the disease onset. In the sera of 40 patients examined 6-7 months after the disease onset IgM-antibodies to WNF virus were absent (dilution 1:50), IgG ones were detected in 35 patients (87.5%). 18 months after the disease IgG-antibodies were identified in 23 of 26 convalescents (88.5%). The serum samples obtained in autumn 1999 and May-June 2000 contained antihemagglutinins to WNF virus for 6-7 months in all 18 patients. In May-June 2000 the sera were positive in 38 convalescents who had WNF in 1999.     

Importance of recrudescent avian infection in West Nile virus overwintering: incomplete antibody neutralization of virus allows infrequent vector infection

After the acute infection period, birds persistently infected with West Nile virus (family Flaviviridae, genus Flavivirus, WNV) occasionally shed virus into the bloodstream, but these virions normally are inactivated by neutralizing antibody. The current work tested the hypothesis that these host neutralizing antibodies protect mosquito vectors from WNV infection and reevaluated the minimum WNV infectious dose necessary to infect Culex tarsalis Coquillett. To determine whether host antibodies protect mosquitoes from infection, Cx. tarsalis and Culex stigmatosoma Dyar were fed bloodmeals containing avian blood, WNV, and sera with or without WNV-specific neutralizing antibodies. When viral particles were completely bound by antibody, mosquitoes were protected from infection; however, when incompletely bound, WNV titers as low as 10(2.3) plaque-forming units (pfu)/ml resulted in 5% infection. These data indicated that avian antibodies were protective to mosquito vectors and were not dissociated during digestion. Because recrudescent viremias may not attain the same magnitude as initial acute viremias, Cx. tarsalis vector competence was reevaluated focusing on the fate of low-titered bloodmeals. Females were evaluated for vector competence after ingesting bloodmeals containing 10(2.2), 10(3.4), 10(4.5), 10(5.5), or 10(6.5) WNV pfu/ml. Infection increased with bloodmeal titer, with 1% of the mosquitoes ingesting 10(3.4) pfu/ml and 45% of the mosquitoes ingesting 10(6.5) pfu/ml developing disseminated infections. The incomplete neutralization of recrudescent virus may be sufficient to infect a low proportion of competent blood-feeding Culex mosquitoes and perhaps allow persistently infected birds to provide a mechanism for arbovirus overwintering.