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In the western United States, this virus may have been mediated via migrating infected birds from southern South America, where it reemerged most recently in 2002., 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).

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.

Changes in the time to first bite (TFB) and the bloodfeeding behavior of adult female Aedes aegypti (L.) mosquitoes following dissemination of Sindbis virus (SINV) were observed after exposure to repellents with the active ingredients (AI) DEET, picaridin, 2-undecanone (2-U), and oil of lemon eucalyptus. Dissemination of SINV significantly decreased (P0.0001) the TFB of DEET (15%) and picaridin (15%) by 46% and 37%, respectively. Significant (P0.0001) changes in activation, probing, and engorgement times were observed in SINV infected mosquitoes after exposure to the four repellents compared to uninfected mosquitoes. Taken together, a decrease in TFB and time to complete the four bloodfeeding stages will lessen the prey-status, and enhance both the chances of mosquito survival and arbovirus transmission.

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.

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.

Disease transmission forecasts can help minimize human and domestic animal health risks by indicating where disease control and prevention efforts should be focused. For disease systems in which weather-related variables affect pathogen proliferation, dispersal, or transmission, the potential for disease forecasting exists. We present a seasonal forecast of St. Louis encephalitis virus transmission in Indian River County, Florida. We derive an empirical relationship between modeled land surface wetness and levels of SLEV transmission in humans. We then use these data to forecast SLEV transmission with a seasonal lead. Forecast skill is demonstrated, and a real-time seasonal forecast of epidemic SLEV transmission is presented. This study demonstrates how weather and climate forecast skill verification analyses may be applied to test the predictability of an empirical disease forecast model.

We analyzed the prevalence of hemagglutination inhibition (HI) antibodies to St. Louis encephalitis (SLE) virus in wild birds during the 1990 SLE epidemic in Indian River County. The initial presence of SLE HI antibody was associated significantly with modeled drought 15 wk prior, wetting conditions 1 wk prior, and the emergence of the Florida SLE virus vector, Culex nigripalpus, 5 wk prior. Our findings indicated that three factors conspired to create the 1990 epidemic: (1) a large population of susceptible wild birds; (2) severe springtime drought, which facilitated amplification of the SLE virus among the Cx. nigripalpus and a portion of the wild bird population; and (3) continued rainfall and wetting of the land surface in the summer and early fall, which sustained a large, host-seeking Cx. nigripalpus population. The continued biting and reproductive activity of Cx. nigripalpus maintained epizootic transmission throughout the summer and early fall in Indian River County. The high level of SLE virus amplification resulted in spillover transmission to humans. We hypothesize that without the continued reproductive activity of the vector mosquito, brought about by excessive summer and fall wetness, the unprecedented SLE virus amplification and consequent transmission to humans would not have been realized in 1990.

West Nile virus (WNv) has spread through much of the eastern United States following its introduction in 1999, and arrived in Florida in 2001. Prior to its arrival, we anticipated that its transmission cycle was likely to be similar to that of St. Louis encephalitis virus (SLEv). However, high levels of avian mortality have been reported for WNv in the northeastern United States, and it was unknown how this would impact the transmission dynamics of WNv. Simulation models were used to compare the two viruses by considering the impact of bird mortality on the transmission dynamics of arboviruses in south Florida. Transmission models without disease-induced mortality (SLEv) were compared with models including disease-induced mortality (WNv). Disease-induced mortality depressed transmission, eliminating epizootics in two of 14 simulations that were epizootic without the additional mortality. In both models, the most important factor in the likelihood of epizootics was mosquito population size; the mosquito mortality rate was also important. The additional avian mortality altered the factors most important in the size and timing of epizootics, although it did not always directly affect the outcome of the simulations. In some cases, low-level transmission occurred prior to the epizootic peak. When disease-induced avian mortality was included in the simulations, appreciable numbers of dead birds occurred prior to high levels of infection in mosquitoes. This has implications for the use of dead birds as a surveillance tool monitoring the spread and transmission of WNv.

▪ Abstract St. Louis encephalitis virus was first identified as the cause of human disease in North America after a large urban epidemic in St. Louis, Missouri, during the summer of 1933. Since then, numerous outbreaks of St. Louis encephalitis have occurred throughout the continent. In south Florida, a 1990 epidemic lasted from August 1990 through January 1991 and resulted in 226 clinical cases and 11 deaths in 28 counties. This epidemic severely disrupted normal activities throughout the southern half of the state for 5 months and adversely impacted tourism in the affected region. The accurate forecasting of mosquito-borne arboviral epidemics will help minimize their impact on urban and rural population centers. Epidemic predictability would help focus control efforts and public education about epidemic risks, transmision patterns, and elements of personal protection that reduce the probability of arboviral infection. Research associated with arboviral outbreaks has provided an understanding of the strengths and weaknesses associated with epidemic prediction. The purpose of this paper is to review lessons from past arboviral epidemics and determine how these observations might aid our ability to predict and respond to future outbreaks.

Four general frequencies of human St. Louis encephalitis (SLE) virus (epidemic, focal, sporadic, and no transmission) occurred in Florida between 1990 and 1999. An epidemic with 226 clinical cases and 11 deaths was reported from 28 Florida counties between July 1990 and January 1991. During the autumn of 1993, a focal outbreak was reported from Lee (5 cases) and Collier (3) Counties in southwest Florida. During the autumn of 1997, sporadic transmission to nine humans was reported from five Florida counties (Brevard [1 case], Polk [3], Charlotte [1], Lee [2], and Palm Beach [2]). Human infection with SLE virus depends on a number of variables that drive virus transmission. These include vector, virus, and avian host abundance, and meteorological events, especially rainfall. We monitored the abundance and serological status of wild avian amplification hosts, virus isolations from Culex nigripalpus Theobald females, and SLE virus transmission to sentinel chickens during 1990, 1993, and 1997. The epidemic of 1990 was characterized by conditions that produced an unusual abundance of vector mosquitoes and avian amplification hosts early in the year. We propose that epidemics may result when a specific combination of biotic and abiotic conditions favor SLE virus minimum field infection rates that approach 1:1,000 in Cx. nigripalpus vectors.

The Report “Emerging vectors in the Culex pipiens complex” by D. M. Fonseca et al. (5 Mar., p. [1535][1]) advances sweeping extrapolations concerning mosquito blood-feeding behavior and West Nile virus (WNV) transmission. These findings led Fonseca to conclude ([1][2]) that a European

From January 1990 through March 1991, 189 armadillos (Dasypus novemcinctus) were collected from Brevard and Glades Counties in southern Florida (USA). The sera were analyzed for hemagglutination-inhibition (HAI) antibodies against St. Louis encephalitis (SLE) and eastern equine encephalomyelitis (EEE) viruses. None of the armadillos had detectable HAI antibody to EEE virus, but 59 (31%) had antibodies against SLE virus. Sera from 31 of the HAI-positive armadillos contained significant levels of neutralizing (NT) antibody to SLE virus. Armadillos captured during the 1990 SLE human epidemic in south Florida had a greater prevalence of HAI and NT antibody to SLE virus than did animals captured before the start of the epidemic. This is evidence that armadillos were fed on by mosquitoes infected with SLE virus. We propose that armadillos may be involved in the SLE amplification and transmission cycles in Florida.

The classic approach to vector control where large tracts of land are treated with an insecticide has many shortcomings. These include high cost, chemical resistance of target species to many of the widely used insecticides, a lack of public acceptance, and the detrimental effect of sprays on nontarget species. Removal trapping, the use of visual, auditory, and olfactory attractants to lure target species into small areas where they are killed, has recently received well-deserved attention as a possible alternative to the broadcast application of chemicals for vector control. We briefly review the histories of four successful removal trapping programs; Hippelates eye gnats in the United States, tsetse flies in Africa, Stomoxys calcitrans flies in Australia, and tabanids in the United States. We then review the future prospects of removal trapping and evaluate its potential as a viable method for vector control.

Mosquito-borne viruses (arboviruses) have the capacity to cause widespread epidemics in humans, as well as epizootics in domestic animals and wildlife. These epidemics and epizootics can be extremely costly and disruptive. Because most mosquito-borne arboviruses can cause encephalitis and encephalomyelitis in host vertebrates, healthcare issues associated with human infections can result in significant economic impact on local economies and health care providers (Villari et al., 1995). Likewise, the loss of domestic animals to encephalitis infection can produce significant adverse financial impacts on local agricultural industries (Wilson et al., 1986; Anon, 2003). Recent mosquito-borne epidemics caused by West Nile virus (family Flaviviridae, genus Flavivirus, WNV) in North America and Chikungunya virus (family Togaviridae, genus Alphavirus, CHIKV) in the islands of the Indian Ocean are examples of how mosquito-borne viruses can severely impact local, regional, and national economies. All mosquito-borne virus disease transmission cycles are driven by four factors: 1) the annual cycle of the pathogen (the specific arbovirus), 2) the annual cycle of the mosquito vector, 3) the annual cycle of the amplification, reservoir, and secondary vertebrate hosts that are infected with a specific viral pathogen, and 4) the environmental factors that drive each biological cycle independently and at times cause these three cycles to synchronize in ways that produce epidemics and epizootics (Day, 2001). Because three biological cycles, pathogen, vector and host, inform the transmission dynamics of a specific arboviral disease, surveillance protocols have been developed to monitor each of these cycles in order to estimate the local risk of arboviral transmission for a specific region. For example, it is possible to use sentinel animals to detect and measure the spatial and temporal distributions of specific arboviruses. Sentinel chickens, pheasants, quail, pigeons, and hamsters have been successfully used to monitor the transmission of St. Louis encephalitis virus (family Flaviviridae, genus Flavivirus, SLEV), WNV, eastern equine encephalitis virus (family Togaviridae, genus Alphavirus, EEEV), and Venezuelan equine encephalitis virus (family Togaviridae, genus Alphavirus, VEEV). Likewise, competent mosquito vectors can be identified for each mosquito-borne arbovirus transmission cycle. Once regional mosquito vectors have been identified, their populations can be tracked and important factors such as abundance, population age structure, and infection status can be monitored and used to

Eastern equine encephalomyelitis virus (EEEV) has been a low-frequency, but serious human and veterinary health problem. Increased frequency of this mosquito-borne virus is anticipated as wetlands are maintained and re-established. Control of EEEV has depended on mosquito abatement in response to increasing frequency of EEEV in the environment. A coupled reverse transcription/polymerase chain reaction assay was designed to rapidly, sensitively, and specifically detect EEEV RNA. The assay successfully detected the viral RNA in a single-blind study of a set of field samples composed of either pooled mosquitoes or bird tissue. These results suggest that it would be practical to use this assay for deciding when and where to implement mosquito abatement.

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.

Sentinel chicken flocks monitored arboviral transmission at 12 sites in Sarasota County, Fla., from 1978 through 1988. During this time, 3,452 serum samples were analyzed for hemagglutination inhibition antibody to St. Louis encephalitis virus (SLE) and eastern equine encephalomyelitis virus (EEE). Overall, 91 chickens seroconverted to SLE and three to EEE. The intensity of SLE transmission showed considerable year-to-year variation. Chickens from south-county flocks were more likely to seroconvert to SLE virus than chickens maintained at the north-county sites. The SLE seroconversion rate for midcounty sentinels was intermediate between north- and south-county sites. Chickens were caged in four habitat types: oak hammocks, slash pine-saw palmetto areas, pastures, and urban-residential areas. There was no significant difference in SLE seroconversion rates between habitats.

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.

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.

Using a dynamic hydrology model, we simulated land surface wetness conditions at 42 sites in 28 counties in southcentral Florida from 1990 to 1998 and compared these simulations with the incidence of human cases of St. Louis encephalitis (SLE) within these counties. Within counties, drought four months prior and wetting one-half month prior were significantly associated with human cases of SLE. Simulated land surface wetness conditions resolved transmission loci in both space and time, and May drought was significantly associated with the subsequent occurrence of human SLE cases. These findings are consistent with previous results associating simulated land surface wetness conditions with the transmission of SLE virus as measured in sentinel chickens, and support our working hypothesis that springtime drought facilitates SLE virus amplification in mosquito and wild bird populations.

Two simulation models were used to investigate the epidemiology of St. Louis encephalitis virus (SLEv) in south Florida, one including sentinel hosts (chickens) and amplification hosts (wild birds), while the other one included age structure in the amplification host population. The overall population size of the vector, Culex nigripalpus, was a major factor in the likelihood of epizootics for both models, but the seasonal dynamics of the vector alone did not explain variation in transmission. Interactions between seasonal factors in the mosquito and reproduction in the wild amplification avian hosts appeared to be important in the likelihood of epizootics. Biased feeding between sentinel and amplification hosts affected the time course of virus prevalence and may have implications for the interpretation of sentinel data. The time of virus introduction strongly affected the timing of outbreaks but did not affect the likelihood of epizootics. In most cases, the outbreak occurred immediately after virus introduction; however, in some cases the outbreak was delayed until the mosquito population increased. This has implications for the timing of control strategies directed against mosquito populations.

A widespread epidemic of St. Louis encephalitis (SLE) virus was reported from central Florida from late July through December 1990 with 226 clinical cases and 11 deaths in humans. The abundance of blood-fed and gravid Culex nigripalpus Theobald females, the vector of SLE in Florida, was monitored at a resting site in Indian River County, FL, for the 3 yr before the epidemic, the epidemic year, and for 1 yr following the epidemic. Each study year was divided into four 3-mo periods that depict the subtropical SLE transmission cycle; a January-March maintenance phase, an April-June amplification phase, a July-September early transmission phase, and an October-December late transmission phase. The abundance of blood-fed and gravid females was significantly greater during the amplification, early transmission, and late transmission phases of the 1990 SLE epidemic when compared with the 3 pre-epidemic years. Throughout the study, Cx. nigripalpus blood feeding and oviposition was driven by epic (> 50 mm) rainfall events. The abundance of gravid females decreased, whereas the number of blood-fed females increased following heavy rainfalls. Epizootic and epidemic transmission of SLE to sentinel chickens and humans in Indian River County was greatest immediately following heavy rainfalls that synchronized Cx. nigripalpus oviposition and blood feeding.

Literature on arthropod-borne diseases has traditionally supported the notion that mosquito vectors maintain a feeding duality that includes vertebrate blood meals for egg development and sugar meals from plants for the synthesis of flight and survival energy reserves. Aedes aegypti was found to deviate from that feeding pattern by obtaining a reproductive advantage when feeding only on human blood. Female mosquitoes fed human blood alone had a greater net replacement rate and intrinsic rate of growth during all phases of their reproductive life than conspecifics fed human blood plus sucrose. Feeding frequently on human hosts during each gonotrophic cycle is necessary to avoid death due to starvation and increases exponentially the spread of Ae. aegypti-borne disease. Our results help explain why Ae. aegypti is such an unusually efficient vector of human disease; frequent biting of humans results in a high reproductive rate for vectors as well as the viruses they transmit.

From 1987 through 1991, blood samples were collected from 10 species of small mammals in Indian River Country, Florida (USA). Sera from 1,347 animals were analyzed for hemagglutination-inhibition (HI) antibody to St. Louis encephalitis (SLE) and eastern equine encephalitis (EEE) viruses. Of these, 75 (5.6%) were positive for HI antibody to SLE virus and 121 (9.0%) were positive for EEE antibody. Sera from five mammalian species were tested for neutralizing (NT) antibody to SLE, EEE, Highlands J (HJ a member of the western equine encephalitis virus complex), or Everglades (EVE, a member of the Venezuelan equine encephalitis complex) viruses. By serum neutralization tests, 26 (46%) of 57 had SLE antibodies, 14 (24%) of 58 had EEE antibodies, two (3.2%) of 63 had HJ antibodies, and 9 (14%) of 63 had EVE antibodies. One Sigmodon hispidus and one Peromyscus gossypinus had NT antibodies both to EEE and HJ viruses. Blood samples from 512 mammals were tested for virus. Isolations of one EVE virus and two unidentified arenaviruses were made from P. gossypinus and one EVE virus isolate was made from a S. hispidus.

From November 1993 through January 1995, sera were collected from 59 domestic emus (Dromaius novaehollandiae) at a ranch in Palm Beach County, FL and tested for antibody evidence of arboviral infection. Hemagglutination inhibition (HI) and neutralizing (NT) antibodies to St. Louis encephalitis (SLE) virus were identified in sera collected each year. In addition, HI and NT antibodies to eastern equine encephalitis (EEE) virus and NT antibodies to western equine encephalitis (WEE) virus were detected in emus imported to and then maintained at the ranch. Neither of the equine encephalitis viruses are common in Palm Beach County and many of the EEE and WEE antibody-positive emus were imported from other states prior to vaccination. Emus imported from California, Louisiana and Texas had evidence of naturally acquired antibodies to EEE and WEE viruses. This observation underscores the potential threat of arboviral introduction by infected vertebrates and emphasizes the importance of instituting quarantine procedures to regulate the transport of hosts that may be infected with arboviral agents.

A method involving injection of anti-estradiol into mice and subsequently assaying this immunoglobulin in the blood meal radio-immunologically was developed to mark conspecific animals for mosquito feeding experiments. Mosquitoes released into cages containing 1 uninfected mouse and 1 mouse infected with either Plasmodium chabaudi or P. berghei engorged almost exclusively on the infected animal. Engorgement success depended on rodent defensive behavior, which was significantly modified by malaria infection. Mosquitoes successfully obtained blood from P. chabaudi -infected mice on day 9–13 postinfection. Mice infected with P. berghei were most vulnerable to mosquito engorgement on day 6–8 and 17–20 postinfection. When cages contained 2 uninfected mice, mosquitoes rarely engorged. Mosquitoes in cages containing 2 malaria-infected mice had engorgement profiles similar to those in cages containing an infected and uninfected mouse.

Laboratory mice infected with rodent malaria (Plasmodium berghei or P. chabaudi) or St. Louis encephalitis virus (SLE) were not hyperthermic during the infection period. However, all infected animals displayed pathogen-specific periods of hypothermia. Hamsters infected with P. berghei were hyperthermic on day 7 postinfection (PI) but became hypothermic on day 8 PI and remained so until death, approximately 20 days PI. Body temperatures of mice infected with P. yoelii were not significantly different from those of uninfected control mice. Mosquitoes (Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus) successfully engorged on restrained, uninfected mice, but were unable to engorge on unrestrained, uninfected mice due to the natural antimosquito behavior of the healthy rodents. Mosquitoes successfully engorged on unrestrained, malaria or SLE infected mice only during certain pathogen-specific periods of infection, but were able to engorge on all restrained, infected mice throughout the infection period regardless of the animal's body temperature. Daily activity patterns of malaria infected mice followed pathogen-specific profiles which closely conformed to the observed mosquito-engorgement profiles.

Mice infected with Plasmodium berghei, P. chabaudi, or P. yoelii became lethargic and ceased to display normal antimosquito behavior. Periods of reduced defensiveness corresponded with maximum mosquito engorgement and with periods of maximum gametocyte infectivity to mosquitoes. Increased feeding success of mosquitoes during periods of peak gametocyte infectivity may be important to the natural maintenance of these malaria parasites.