Can Horton hear the whos? The importance of scale in mosquito-borne disease.

The epidemiology of vector-borne pathogens is determined by mechanisms and interactions at different scales of biological organization, from individual-level cellular processes to community interactions between species and with the environment. Most research, however, focuses on one scale or level with little integration between scales or levels within scales. Understanding the interactions between levels and how they influence our perception of vector-borne pathogens is critical. Here two examples of biological scales (pathogen transmission and mosquito mortality) are presented to illustrate some of the issues of scale and to explore how processes on different levels may interact to influence mosquito-borne pathogen transmission cycles. Individual variation in survival, vector competence, and other traits affect population abundance, transmission potential, and community structure. Community structure affects interactions between individuals such as competition and predation, and thus influences the individual-level dynamics and transmission potential. Modeling is a valuable tool to assess interactions between scales and how processes at different levels can affect transmission dynamics. We expand an existing model to illustrate the types of studies needed, showing that individual-level variation in viral dose acquired or needed for infection can influence the number of infectious vectors. It is critical that interactions within and among biological scales and levels of biological organization are understood for greater understanding of pathogen transmission with the ultimate goal of improving control of vector-borne pathogens.

Environmental risk factors associated with West Nile virus clinical disease in Florida horses.

The objective of this study was to examine the extrinsic risk factors of West Nile virus (WNV) clinical disease in Florida horses as established from confirmed and negative horses tested within the state from 2001 to 2003. An Arboviral Case Information Form (ACF) was submitted by a referring veterinarian at the time of testing to the Florida Department of Agriculture and Consumer Services on every horse suspected of a viral encephalitis in Florida. A follow-up survey that focused on arbovirus prevention and farm ecology was created and mailed to the owner of each tested horse. Data from the follow-up survey indicated peak WNV prevalence in the late summer months in Florida. Quarter horses were the most commonly affected breed. The WNV vaccine was highly protective and natural water on the property also had a protective association. Factors that increased the risk of WNV to horses were the use of fans and a stable construction of solid wood or cement. Some risk indicators were dead birds on the property and other ill animals on the property. Data from this retrospective study have helped identify factors associated with WNV transmission in equines in Florida. Horses that have not been vaccinated and show clinical signs of arboviral infection from June to November should be tested for WNV. Horses that have been vaccinated and show clinical signs should be tested when the vaccination was administered within 1 month or greater than 6 months prior to the onset of clinical symptoms associated with WN infection.

A simple method for sampling indoor-resting malaria mosquitoes Anopheles gambiae and Anopheles funestus (Diptera: Culicidae) in Africa.

Sampling indoor resting African malaria vectors is traditionally done by hand catches with oral or mechanical aspirators and pyrethrum spray catches (PSCs). In this study, we designed and briefly evaluated an inexpensive but practical alternative by using a cloth resting box or wicker resting basket and a ceiling net. Evaluations were performed in greenhouse and field situations in rural Kenya by comparing capture rates of Anopheles gambiae s.l. and Anopheles funestus (Giles) in these traps to hand collections and PSCs. A resting box and a ceiling net when used together collected more mosquitoes than a single collector using a hand-held aspirator but only one-third the number collected by PSCs. At sites where PSCs are impractical, a resting box and ceiling net can be effectively used as an alternative to hand catches in malaria surveillance.

Nonenzymatic glucosylation of serum proteins and hemoglobin: response to changes in blood glucose levels in diabetic rats.

The relationship between concentrations of blood glucose and nonenzymatically glucosylated serum proteins was studied in rats with alloxan-induced diabetes of varying severity. Fasting serum glucose correlated strongly with both glucosylated albumin (r = 0.91, P less than 0.001) and glucosylated serum protein (r = 0.93, P less than 0.001). The relative rates of response of serum protein and hemoglobin glycosylation to changes in blood glucose were also compared. Following withdrawal of insulin from diabetic rats, the half-times to reach new steady state levels of blood glucose, glucosylated serum proteins, and glycohemoglobins were about 2, 3, and 8 days, respectively. Similarly, on reinstitution of insulin therapy, the half-times for these same indices to return to baseline values were 2, 3.5, and 15 days, respectively. Changes in glucosylated albumin were more sensitive than glycohemoglobins to changes in serum glucose, consistent with the observation that albumin was glucosylated at about 10 times the rate for hemoglobin in incubations in vitro. These data indicate that glucosylated serum protein measurements can serve as a sensitive, short-term integrator of blood glucose homeostasis in diabetes.

Nonenzymatic glucosylation of serum proteins in diabetes mellitus.

The extent of nonenzymatic glucosylation of serum protein in control and diabetic subjects was measured by a chemical procedure using thiobarbituric acid. A mean value of 0.81 (+/- 0.21 SD) nmol glucose per milligram serum protein was observed in the control group. Diabetics displayed elevated levels of glucosylated serum proteins, up to 4 nmol glucose per milligram protein. Glucosylation of serum protein correlated strongly with fasting blood sugar (r = 0.71), percent hemoglobin A1 (r = 0.79), and percent glucosylated albumin (r = 0.99). There was no overlap between control and diabetic groups, i.e., within 3 SD of the mean of controls. These studies indicate that the assay for glucosylated serum protein appears to be an especially sensitive indicator of the degree of hyperglycemia in diabetes.

Chemical odorant of colonial seabird repels mosquitoes.

The crested auklet, Aethia cristatella, emits a class of aldehydes shown to be potent invertebrate repellents when used by heteropterans against their predators. Our aim was to determine the efficacy of these aldehydes against mosquitoes in the laboratory. Synthetic analogues of the auklet odorant were strongly repellent to mosquitoes in controlled laboratory trials. Furthermore, the efficacy was similar to previous reports for commercial mosquito repellents. These results, in combination with a previously published study, show that constituents of the aldehyde odorant are broad spectrum in efficacy against ectoparasitic arthropods of birds. Our report is the first empirical evidence for an endogenous mosquito repellent in birds.

Vector control by removal trapping.

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.

A rapid diagnostic assay for eastern equine encephalomyelitis viral RNA.

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.

A fitness advantage for Aedes aegypti and the viruses it transmits when females feed only on human blood.

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.

Simulation studies of St. Louis encephalitis virus in south Florida.

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.

Simulation studies of St. Louis encephalitis and West Nile viruses: the impact of bird mortality.

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.

Frequency of Saint Louis encephalitis virus in humans from Florida, USA: 1990-1999.

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.

First record of Amblyomma auricularium (Acari: Ixodidae) in the United States.

Amblyomma auricularium (Conil) was collected from an armadillo in Glades County, FL. Two females and 11 males were collected. This is the first record of this species in the United States.

Transmission patterns of St. Louis encephalitis and eastern equine encephalitis viruses in Florida: 1978-1993.

Sentinel chickens were maintained at field sites in 40 Florida counties for varying periods between 1978 and 1993. For each county, the total number of St. Louis encephalitis virus (SLE) or eastern equine encephalitis (EEE) virus seroconversions were divided by the number of chickens exposed to calculate a mean annual seroconversion rate. These rates were used to evaluate the annual and geographical distributions of these viruses within Florida. For SLE, the rates in counties that reported human SLE cases during a widespread epidemic in 1990 were compared with adjusted mean annual seroconversion rates calculated by excluding the epidemic year seroconversion data. In general, actual rates were higher than adjusted rates in counties where human SLE cases were reported in 1990. Similar calculations were made for EEE virus that was unusually abundant in northern Florida during the spring and summer of 1991. In general, EEE virus was distributed in the panhandle and northern regions and SLE virus in the central and southern regions of the state. However, this distribution was not exclusive, and during years of high transmission, either virus can extend beyond its normal range. The annual distribution of SLE and EEE viruses was sporadic, ranging from years with little detectable activity to years with widespread, high-level transmission.

Blood feeding and oviposition by Culex nigripalpus (Diptera: Culicidae) before, during, and after a widespread St. Louis encephalitis virus epidemic in Florida.

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.

Avian serology in a St. Louis encephalitis epicenter before, during, and after a widespread epidemic in south Florida, USA.

Blood and serum from 3,915 wild and domestic birds (2,590 resident, 139 migrant, and 1,186 captive), representing 56 species collected in central Florida from 1989 through 1997, were analyzed for evidence of St. Louis encephalitis (SLE) virus transmission. All sera were tested for SLE hemagglutination inhibition (HI) antibody. Selected sera and bloods were tested for SLE neutralizing (NT) antibody and virus. The reproductive success of resident birds was highest from 1990-1992 and lowest from 1994-1997. Transmission of SLE to resident birds, especially mourning doves (Zenaida macroura), peaked during the summer of 1990, a year during which a widespread SLE epidemic was recorded in central Florida. The SLE antibody-positive resident birds 1st appeared during September of the epidemic year. Some SLE, HI antibody-positive resident birds were captured throughout 1991, but only 5% were yearlings, compared with 36% in 1990. By 1993, wild resident birds expressing HI and NT antibodies to SLE had nearly disappeared. None of the migrant birds tested were SLE-positive. Sentinel chickens maintained in Indian River County during the epidemic year seroconverted to SLE starting in early July with peak seroconversion rates in August, September, and October. High (> or = 50%) SLE seroconversion rates in sentinel chickens preceded those in wild birds by 10 wk and preceded peak human SLE transmission by at least 8 wk. Major SLE epidemics in south Florida depend on abundant wild bird populations, especially during the amplification phase of the transmission cycle. We propose that hard winter freezes along the temperature-subtropical climatic zone interface in central Florida, at approximately 27 degrees 30' North Latitude, opens foraging and nesting habitats for ground-feeding birds, resulting in high reproductive success and an abundance of seronegative individuals that rapidly amplify the SLE later in the year.

Ultrastructure of the eggs of seven flea species (Siphonaptera).

Eggs of seven flea species representing five families, Ctenocephalides felis (Bouché) (Pulicidae), Orchopeas leucopus (Baker) and Aetheca wagneri (Baker) (Ceratophyllidae), Conorhinopsylla stanfordi Stewart and Epitedia faceta (Rothschild) (Hystrichopsyllidae), Sternopsylla distincta texana (C. Fox) (Ischnopsyllidae), and Craneopsylla minerva (Rothschild) (Stephanocircidae), were examined with a scanning electron microscope and are described from the resulting micrographs. In the Pulicidae and Ceratophyllidae the individual outer chorionic cells are not visible; the egg surfaces are covered uniformly with small, nodular tubercles. In contrast, the Hystrichopsyllidae and Ischnopsyllidae have a prominent, raised reticulum, very heavily constructed in the latter, delineating the chorionic cells. The single representative of the Stephanocircidae has only a faintly embossed chorionic reticulum. All of the species examined except E. faceta and C. minerva (both with a single micropylar opening) have multiple micropyles at the posterior end. C. felis, O. leucopus, A. wagneri, and C. stanfordi also have multiple aeropyles at the anterior end (these are lacking in S. d. texana). Lateral aeropyles, consisting of small groups of pores often associated with small mounds in the chorion, are present (maximum of two seen on any one egg) in all species except E. faceta, C. stanfordi, and C. minerva.

Distribution of St. Louis encephalitis viral antibody in sentinel chickens maintained in Sarasota County, Florida: 1978-1988.

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.

Reproductive fitness and survivorship of Aedes aegypti (Diptera: Culicidae) maintained on blood, with field observations from Thailand.

Daily survivorship and fecundity of Aedes aegypti mosquitoes were monitored for cohorts provided with five nutritional treatments: water, 2% sucrose, blood from a live chicken plus water, blood plus 2% sucrose, and blood alone. The median mortality time (LT50) for these females was 6, 54, 16, 12, and 29 d, respectively. There was no significant difference in the number of eggs laid by females in any of the treatments containing a host. Females maintained on blood alone laid as many or more eggs during their lifetime as females with access to sugar who had a greater life expectancy. Males maintained on sugar alone survived significantly longer than those in any of the other treatments. Large- and small-bodied, sugar-starved Ae. aegypti females that were marked, released, and recaptured in a Thai village survived as well as the replicate cohorts that received sugar or sugar plus blood during the 36 +/- 12 h period from emergence until their release. These results indicate that Ae. aegypti females live longer in the laboratory if they are provided a source of carbohydrate, but the increased survival associated with sugar feeding does not increase reproductive success.

Rainfall-directed oviposition behavior of Culex nigripalpus (Diptera: Culicidae) and its influence on St. Louis encephalitis virus transmission in Indian River County, Florida.

Resting Culex nigripalpus Theobald were collected with a ground aspirator three times a week in Indian River County, Fla., from January 1985 through December 1987. The number of green (newly emerged), empty, blood-fed, and gravid females in each collection was counted; up to 100 empty females were dissected to estimate the abundance of empty, parous females in the population. In general, this species was uncommon during the dry season from January through June but increased during the wet season in July and remained abundant through early winter. During the wet season, green, parous, and gravid females were periodically abundant. Time series analysis demonstrated that green and parous mosquitoes were positively associated through time and that gravid females were associated negatively with daily rainfall. The strongest correlations were recorded in 1985, a year during which St. Louis encephalitis virus was detected with increased frequency in Indian River County. Laboratory experiments in a large outdoor wind tunnel confirmed that high relative humidity was important for the initiation and successful completion of oviposition by gravid, field-collected Cx. nigripalpus mosquitoes. Thus, intermittent rainfall during autumn may serve to delay oviposition, increase longevity, synchronize blood feeding, and enhance transmission of St. Louis encephalitis virus by infected mosquitoes in the field.