Reemergence of St. Louis Encephalitis Virus in the Americas.

We summarize and analyze historical and current data regarding the reemergence of St. Louis encephalitis virus (SLEV; genus Flavivirus) in the Americas. Historically, SLEV caused encephalitis outbreaks in the United States; however, it was not considered a public health concern in the rest of the Americas. After the introduction of West Nile virus in 1999, activity of SLEV decreased considerably in the United States. During 2014-2015, SLEV caused a human outbreak in Arizona and caused isolated human cases in California in 2016 and 2017. Phylogenetic analyses indicate that the emerging SLEV in the western United States is related to the epidemic strains isolated during a human encephalitis outbreak in Córdoba, Argentina, in 2005. Ecoepidemiologic studies suggest that the emergence of SLEV in Argentina was caused by the introduction of a more pathogenic strain and increasing populations of the eared dove (amplifying host).

Factors That Influence the Transmission of West Nile Virus in Florida.

West Nile virus (WNV) was first detected in North America in New York City during the late summer of 1999 and was first detected in Florida in 2001. Although WNV has been responsible for widespread and extensive epidemics in human populations and epizootics in domestic animals and wildlife throughout North America, comparable epidemics have never materialized in Florida. Here, we review some of the reasons why WNV has yet to cause an extensive outbreak in Florida. The primary vector of mosquito-borne encephalitis virus in Florida is Culex nigripalpus Theobald. Rainfall, drought, and temperature are the primary factors that regulate annual populations of this species. Cx. nigripalpus is a competent vector of WNV, St. Louis encephalitis virus, and eastern equine encephalitis virus in Florida, and populations of this species can support focal amplification and transmission of these arboviruses. We propose that a combination of environmental factors influencing Cx. nigripalpus oviposition, blood-feeding behavior, and vector competence have limited WNV transmission in Florida to relatively small focal outbreaks and kept the state free of a major epidemic. Florida must remain vigilant to the danger from WNV, because a change in these environmental factors could easily result in a substantial WNV epidemic rivaling those seen elsewhere in the United States.

2015 AMCA Memorial Lecture Honoree: Dr. Richard Floyd Darsie, Jr.

Richard Floyd Darsie, Jr. (1915-2014) is the 2015 American Mosquito Control Association Memorial Lecture Honoree. He was one of the greatest mosquito taxonomists of the 20th century and died peacefully on April 10, 2014, in Grove City, PA, at the age of 99 after a professional career that spanned eight decades. Dick's broad areas of interest and training made him a versatile scientist, teacher, and researcher. His intense interest in adult and immature mosquito morphology and taxonomy, as well as mosquito distribution and bionomics, started early in his career at two early academic postings: Franklin and Marshall College (1949-54) and the University of Delaware (1954-62). Dick would take his mosquito interests with him to postings and research projects around the world: Nepal, the Philippines, Atlanta, El Salvador, Guatemala, Fort Collins, South Carolina, Argentina, and Florida. His travels and studies would make him an international expert on mosquito taxonomy. Dick's legacy lives on in the hundreds of students from across the globe who learned mosquito identification skills from this world-renowned mosquito taxonomist. All will proudly profess that, "I learned mosquito identification from Dr. Darsie." And that is all that is needed to prove one's credentials in the field, learning the art from the best there is.

Sindbis virus infection alters blood feeding responses and DEET repellency in Aedes aegypti (Diptera: Culicidae).

Aedes aegypti (L.) (Diptera: Culicidae) female mosquitoes infected systemically with Sindbis virus (SINV) took longer than uninfected mosquitoes to locate and fully engorge on blood. On days 7 and 14 postexposure, blood feeding took 1.3 and 1.5 times longer in mosquitoes with a disseminated SINV infection, respectively. SINV dissemination did not affect the average weight of unfed Ae. aegypti, but did result in a 10 and 12% increase in blood imbibed compared with mosquitoes without a positive SINV dissemination and non-SINV-exposed mosquitoes, respectively. Ae. aegypti mosquitoes with a disseminated SINV infection fed an average of 4 h sooner than uninfected mosquitoes when offered a bloodmeal contained inside a DEET (N,N-diethyl-3-methylbenzamide) saturated (30%) bovine sausage casing. Together, these results indicate that behavioral changes in mosquito host-seeking, blood feeding and sensitivity to DEET occurred in mosquitoes after SINV infection and dissemination.

Ecology of Eastern Equine Encephalitis Virus in the Southeastern United States: Incriminating Vector and Host Species Responsible for Virus Amplification, Persistence, and Dispersal.

Eastern equine encephalitis virus (EEEV; family Togaviridae, genus Alphavirus) is a mosquito-borne pathogen found in eastern North America that causes severe disease in humans and horses. The mosquito Culiseta melanura (Coquillett) (Diptera: Culicidae) is the primary enzootic vector of EEEV throughout eastern North America while several mosquito species belonging to diverse genera serve as bridge vectors. The ecology of EEEV differs between northern and southern foci, with respect to phenology of outbreaks, important vertebrate hosts, and bridge vector species. Active transmission is limited to roughly half of the year in northern foci (New York, New Hampshire, Massachusetts, Connecticut), while year-round transmission occurs in the southeastern region (particularly Florida). Multiple phylogenetic analyses indicate that EEEV strains circulating in northern foci are likely transported from southern foci by migrating birds. Bird species that overwinter or migrate through Florida, are bitten by Cs. melanura in late spring, and arrive at northern breeding grounds in May are the most likely candidates to disperse EEEV northward. Available data indicate that common yellowthroat and green heron satisfy these criteria and could serve as virus dispersers. Understanding the factors that drive the phenology of Cs. melanura reproduction in the south and the timing of avian migration from southern foci could provide insight into how confluence of these biological phenomena shapes outbreaks of EEE throughout its range. This information could be used to develop models predicting the likelihood of outbreaks in a given year, allowing vector control districts to more efficiently marshal resources necessary to protect their stakeholders.

Altered response to DEET repellent after infection of Aedes aegypti (Diptera: Culicidae) with Sindbis virus.

To determine whether a Sindbis virus (family Togaviridae, genus Alphavirus, SINV) infection in Aedes aegypti (L.) (Diptera: Culicidae) affected its response to the repellent DEET, we orally exposed Ae. aegypti to an artificial bloodmeal containing SINV or diluent and then allowed to feed on a 10% sucrose suspension containing 3% DEET. When tested seven or more days after the initial bloodmeal, although none of the diluent-exposed mosquitoes fed on the DEET-sucrose suspension, at least 60% of the SINV-exposed mosquitoes fed on the suspension. When legs from the SINV-exposed mosquitoes were tested to determine dissemination status, 89% of those that fed on the DEET-sucrose suspension contained virus. In contrast, only 34% of the nonfeeders had a disseminated infection. When offered a choice between sucrose with or without DEET, a significantly higher percentage of the SINV-exposed mosquitoes than the control mosquitoes fed on the sucrose containing 3% DEET. Together, these results indicate that mosquitoes with a disseminated SINV infection may be less responsive to DEET than uninfected mosquitoes. Therefore, repellent use may be less effective in deterring infected mosquitoes from biting than previously believed.

Countering a bioterrorist introduction of pathogen-infected mosquitoes through mosquito control.

The release of infected mosquitoes or other arthropods by bioterrorists, i.e., arboterrorism, to cause disease and terror is a threat to the USA. A workshop to assess mosquito control response capabilities to mount rapid and effective responses to eliminate an arboterrorism attack provided recommendations to improve capabilities in the USA. It is essential that mosquito control professionals receive training in possible responses, and it is recommended that a Council for Emergency Mosquito Control be established in each state to coordinate training, state resources, and actions for use throughout the state.

Seasonal Changes of Host Use by Culiseta melanura (Diptera: Culicidae) in Central Florida.

The mosquito Culiseta melanura (Coquillett) is the primary enzootic vector of eastern equine encephalitis virus (EEEV), a zoonotic Alphavirus endemic to eastern North America. In its northern range, Cs. melanura is considered a strict avian biter, transmitting EEEV among susceptible birds in a cycle of enzootic amplification. In its southern range, however, Cs. melanura is more general in host use, feeding heavily upon birds but also reptiles and mammals. The goal of this study was to better understand how host use of Cs. melanura changes throughout the year in Florida, where year-round EEEV transmission is observed. Mosquitoes were sampled in 2018 from nine sites across three central Florida counties. In total, 213 Cs. melanura bloodmeals were identified by PCR consisting of 39 species of birds, reptiles, and mammals. Avian bloodmeals were prominent throughout the year (range = 30-85%), and songbirds were a large portion of identified bloodmeals (37.1%). Reptiles surpassed birds only in spring (April-June), and brown anole (Anolis sagrei Duméril and Bibron, 1837 [Reptilia: Dactyloidae]) was the most commonly detected single host species (22.1% overall). Mammalian bloodmeals were mainly observed in summer, with humans being the most fed on mammal (12.7% overall). This study reveals that in southern foci of EEEV transmission, Cs. melanura host use varies throughout the year with reptiles providing the majority of blood meals in spring (51.3%), and birds are fed on more than other host groups during all other seasons (50.6-70.1%). In addition, feeding on mammals increases during summer months, which may implicate Cs. melanura in epizootic transmission in Florida.

Severe winter freezes enhance St. Louis encephalitis virus amplification and epidemic transmission in peninsular Florida.

Mosquito-borne arboviral epidemics tend to strike without warning. The driving force for these epidemics is a combination of biotic (vector, amplification host, and virus) and abiotic (meteorological conditions, especially rainfall and temperature) factors. Abiotic factors that facilitate the synchronization and interaction of vector and amplification host populations favor epidemic amplification and transmission. In Florida, epidemics of St. Louis encephalitis (SLE) virus (family Flaviviridae, genus Flavivirus, SLEV) have been preceded by major freezes one or two winters before the onset of human cases. Here, we analyze the relationship between severe winter freezes and epidemic SLEV transmission in peninsular Florida and show that there is a significant relationship between the transmission of SLEV and these severe freezes. We propose that by killing cold-sensitive understory vegetation in the mid-peninsular region of Florida, freezes enhance the reproductive success of ground-feeding avian amplification hosts, especially mourning doves and common grackles. In conjunction with other appropriate environmental signals, increased avian reproductive success may enhance SLEV and West Nile (WN) virus amplification and result in SLE and WN epidemics during years when all of the biological cycles are properly synchronized. The knowledge that winter freezes in Florida may enhance the amplification and epidemic transmission of SLE and WN viruses facilitates arboviral tracking and prediction of human risk of SLE and WN infection during the transmission season.

Using hydrologic conditions to forecast the risk of focal and epidemic arboviral transmission in peninsular Florida.

The accurate forecasting and tracking of arboviral transmission is becoming increasingly critical for the early recognition and management of arboviral epidemics. Meteorological factors, especially rainfall and temperature, drive arboviral epidemics, but monitoring rainfall and temperature alone is not predictive of increased levels of vector-borne disease transmission. In Florida, model simulations of water table depth (WTD) provide a measure of drought, and they have been shown to provide an accurate forecast of arboviral transmission. Here, we tracked WTD in two peninsular Florida regions where focal West Nile virus (family Flaviviridae, genus Flavivirus, WNV) transmission was reported during 2004 and 2005. We compared the resulting WTD profiles with historical WTD simulations for Indian River County (IRC), FL, where two peninsular Florida St. Louis encephalitis virus epidemics had their epicenters in 1977 and 1990. In both of the regions where focal WNV transmission was reported during 2004 and 2005, the local WTD profiles approached the 1977 and 1990 IRC WTD profiles; however, differences in the local temporal sequence of hydrologic conditions were observed. These differences seem in part to explain why the focal WNV transmission during 2004 and 2005 failed to reach epidemic levels in peninsular Florida. These findings suggest that hydrologic monitoring, specifically WTD, may help determine the geographic extent, timing, and intensity of WNV transmission. We speculate that a more precise sequence of drought and wetting, including a secondary summer drying and wetting cycle, as occurred in IRC during 1977 and 1990, may provide the optimal hydrologic conditions for the expansion of an arbovirus outbreak from focal to epidemic. This study documents that monitoring hydrologic conditions, along with vector, avian amplification host, and virus population data, increases our ability to track and predict significant levels of arboviral transmission.

Arbovirus transmission by Culex nigripalpus in Florida, 2005.

Understanding the transmission patterns of West Nile and St. Louis encephalitis viruses (family Flaviviridae, genus Flavivirus, WNV and SLEV) could result in an increased ability to predict transmission risk to humans. To examine transmission patterns between vector and host, we trapped mosquitoes in three Florida counties from June to November 2005 by using chicken-baited lard can mosquito traps. These traps were used to monitor for presence of WNV and SLEV in mosquitoes and subsequent transmission of these viruses to chickens. In total, 166,615 female mosquitoes were sorted into 4,009 pools based on species and bloodfed status, and they were tested for presence of WNV and SLEV. Sera from 209 chickens were tested for WNV and SLEV antibodies. We detected eight WNV-positive Culex nigripalpus Theobald mosquito pools; SLEV was not detected in any pools. Six positive pools were collected in August and September from Duval County, one pool in September from Manatee County, and one pool in November from Indian River County. Of the eight chickens potentially exposed to WNV, antibodies were detected in only one chicken, indicating a low rate of transmission relative to the observed mosquito infection rates. Low virus transmission rates relative to infection rates would suggest that using sentinel chicken seroconversion data as a means of arbovirus surveillance may underestimate the prevalence of WNV in the mosquito population. However, using mosquito infection rates may overestimate the risk of arboviral transmission. A variety of factors might account for the observed low level of transmission including a lack of viral dissemination in mosquito vectors.

Redescription of the pupa of Culex restuans and a comparison with Culex nigripalpus.

The pupa of Culex restuans is redescribed in detail with a chaetotaxal table and a full illustration. The chaetotaxy of the pupa of Cx. restuans is compared with that of Cx. nigripalpus, the primary vector of St. Louis encephalitis and West Nile virus in Florida.

Redescription of the pupa of Culex salinarius Coquillett and comparison with Culex nigripalpus Theobald.

The pupa of Culex salinarius is redescribed with updated chaetotaxal nomenclature and a full illustration. The pupal chaetotaxy of Cx. salinarius and the similar species Culex nigripalpus is compared.

Studies of the genus Culex linnaeus in Florida. III. Redescription of the fourth-stage larva of Culex salinarius Coquillett and comparison with that of Cx. nigripalpus Theobald.

The larva of Culex salinarius is completely described and illustrated, except for the mouth parts, and compared with morphologically similar larva of Cx. nigripalpus. At least 8 characters have been found to separate larvae of the two species of which 7 give 90% or higher divergence while the remaining character gives 87% separation.

Interisland dispersal of the black salt marsh mosquito, Ochlerotatus taeniorhynchus (Diptera: Culicidae) in the Florida Keys.

Mark-release-recapture experiments were conducted in 2001 and 2002 to determine whether Ochlerotatus taeniorhynchus, black salt marsh mosquitoes, were dispersing from uninhabited islands in the Key Deer National Wildlife Refuge to inhabited islands within Monroe County, Florida. An estimated 1,658,000 mosquitoes were marked during 2001, and an estimated 300,000 mosquitoes were marked during 2002. Recapture rates were 0.0061% and 0.0117%, respectively. Oc. taeniorhynchus disperse from uninhabited islands to other uninhabited islands and also to inhabited islands, namely, Big Pine Key and No Name Key.

Mosquito Oviposition Behavior and Vector Control.

The burden of gene transfer from one mosquito generation to the next falls on the female and her eggs. The selection of an oviposition site that guarantees egg and larval survival is a critical step in the reproductive process. The dangers associated with ephemeral aquatic habitats, lengthy droughts, freezing winters, and the absence of larval nutrition makes careful oviposition site selection by a female mosquito extremely important. Mosquito species exhibit a remarkable diversity of oviposition behaviors that ensure eggs are deposited into microenvironments conducive for successful larval development and the emergence of the next mosquito generation. An understanding of mosquito oviposition behavior is necessary for the development of surveillance and control opportunities directed against specific disease vectors. For example, Aedes aegypti Linnaeus is the vector of viruses causing important human diseases including yellow fever, dengue, chikungunya, and Zika. The preference of this species to oviposit in natural and artificial containers has facilitated the development of Ae. aegypti-specific surveillance and toxic oviposition traps designed to detect and control this important vector species in and around disease foci. A better understanding of the wide diversity of mosquito oviposition behavior will allow the development of new and innovative surveillance and control devices directed against other important mosquito vectors of human and animal disease.

Drought-induced amplification and epidemic transmission of West Nile virus in southern Florida.

We show that the spatial-temporal variability of human West Nile (WN) cases and the transmission of West Nile virus (WNV) to sentinel chickens are associated with the spatial-temporal variability of drought and wetting in southern Florida. Land surface wetness conditions at 52 sites in 31 counties in southern Florida for 2001-2003 were simulated and compared with the occurrence of human WN cases and the transmission of WNV to sentinel chickens within these counties. Both WNV transmission to sentinel chickens and the occurrence of human WN cases were associated with drought 2-6 mo prior and land surface wetting 0.5-1.5 mo prior. These dynamics are similar to the amplification and transmission patterns found in southern Florida for the closely related St. Louis encephalitis virus. Drought brings avian hosts and vector mosquitoes into close contact and facilitates the epizootic cycling and amplification of the arboviruses within these populations. Southern Florida has not recorded a severe, widespread drought since the introduction of WNV into the state in 2001. Our results indicate that widespread drought in the spring followed by wetting during summer greatly increase the probability of a WNV epidemic in southern Florida.

Host-seeking strategies of mosquito disease vectors.

Disease transmission by arthropods normally requires at least 2 host contacts. During the first, a pathogen (nematode, protozoan, or virus) is acquired along with the blood from an infected vertebrate host. The pathogen penetrates the vector's midgut and infects a variety of tissues, where replication may occur during an extrinsic incubation period lasting 3-30, days depending on vector and parasite physiology and ambient temperature. Following salivary-gland infection, the pathogen is usually transmitted to additional susceptible vertebrate hosts during future probing or blood feeding. The host-seeking strategies used by arthropod vectors can, in part, affect the efficiency of disease transmission. Vector abundance, seasonal distribution, habitat and host preference, and susceptibility to infection are all important components of disease-transmission cycles. Examples of 3 mosquito vectors of human disease are presented here to highlight the diversity of host seeking and to show how specific behaviors may influence disease-transmission cycles. In the African tropics, Anopheles gambiae s.s. is an efficient vector of human malaria due to its remarkably focused preference for human blood. Aedes aegypti is the main vector of dengue viruses in the New and Old World tropics and subtropics. This mosquito has evolved a domestic lifestyle and shares human habitations throughout much of its range. It prospers in settings where humans are its main source of blood. In south Florida, Culex nigripalpus is the major vector of St. Louis encephalitis (SLE) and West Nile (WN) viruses. This mosquito is opportunistic and blood feeds on virtually any available vertebrate host. It serves as an arboviral vector, in part, due to its ability to produce large populations in a short period of time. These 3 host-seeking and blood-feeding strategies make the specialist, as well as the opportunist, equally dangerous disease vectors.

Redescription of the pupa of Culex (Culex) declarator Dyar and Knab (Diptera, Culicidae), with amendments to key to the Culex pupae of the eastern United States.

The pupa of Culex declarator was partially described and illustrated by Lane. This species was recently discovered in Florida for the first time and a series of individual rearings have resulted in a number of pupae and an opportunity to redescribe it fully, with a complete illustration. Amendment to the key to the Culex pupae of the eastern United States is provided.