Spatial-temporal clusters of host-seeking Aedes albopictus, Aedes japonicus, and Aedes triseriatus collections in a La Crosse virus endemic county (Knox County, Tennessee, USA).

A bite from a La Crosse virus (LACV) infected Aedes mosquito can cause La Crosse encephalitis (LACE), which is a neuro-invasive disease that disproportionately affects children under the age of 16 in Southern Appalachia. The three vectors for LACV are Aedes albopictus (Skuse), Ae. japonicus (Theobald), and Ae. triseriatus (Say). Localized maps of the geographic distribution of vectors are practical tools for mosquito management personnel to target areas with high mosquito abundance. This study hypothesized that LACV vectors have unique species-specific spatial and temporal clusters. To test this, 44 sites were identified in Knox County, Tennessee for their land use/type. At each site, host-seeking mosquitoes were collected approximately every other week from May-October 2018. Spatial clusters of host-seeking mosquito collections for each of the three mosquito species were investigated using Kulldorff's spatial scan statistic, specifying a retrospective space-time Bernoulli model. Most vector clusters were identified in south-central Knox County while the seasonality of clusters varied by mosquito species. Clusters of Ae. albopictus were observed throughout the entire study period while clusters of Ae. japonicus and Ae. triseriatus only occurred May-June. The findings indicate that the relative abundance of LACV vectors were more abundant in south-central Knox County compared to the rest of the county. Of interest, these clusters spatially overlapped with previous LACE diagnosed cases. These findings are useful in guiding decisions on targeted mosquito control in Knox County and may be applied to other counties within Southern Appalachia.

Spatiotemporal Co-occurrence of Flanders and West Nile Viruses Within Culex Populations in Shelby County, Tennessee.

West Nile virus (WNV) and Flanders virus (FLAV) can cocirculate in Culex mosquitoes in parts of North America. A large dataset of mosquito pools tested for WNV and FLAV was queried to understand the spatiotemporal relationship between these two viruses in Shelby County, TN. We found strong evidence of global clustering (i.e., spatial autocorrelation) and overlapping of local clustering (i.e., Hot Spots based on Getis Ord Gi*) of maximum likelihood estimates (MLE) of infection rates (IR) during 2008-2013. Temporally, FLAV emerges and peaks on average 10.2 wk prior to WNV based on IR. Higher levels of WNV IR were detected within 3,000 m of FLAV-positive pool buffers than outside these buffers.

Host associations of Dermacentor, Amblyomma, and Ixodes (Acari: Ixodidae) ticks in Tennessee.

From April 2007 to September 2008, 1,793 adult and nymphal ixodid ticks were collected from 49 counties in Tennessee. Six species were identified, including Dermacentor variabilis (Say), Amblyomma americanum (L.), Ixodes texanus (Banks), Ixodes cookei Packard, Ixodes scapularis (Say), and Amblyomma maculatum Koch, from 13 medium- to large-sized mammalian hosts and dragging through vegetation. Raccoons were the most common vertebrate source (198 captures), accounting for 60% of ticks collected. Dermacentor variabilis was the predominant species from raccoons with a prevalence of 92% and mean intensity of 5.3. A. americanum was predominated in white-tailed deer and drags with respective mean intensities of 3.1 and 14.1 and prevalence values of 94%. All tick species were identified between April and August, coinciding with the majority of animal captures. Only A. americanum, I. texanus, and I. cookei were identified from 22 animal captures from November to March. I. texanus and I. cookei were more common in the eastern portions of the state, but this may be a result of higher raccoon captures in those areas. Only four specimens of I. scapularis were collected in this study, which may reflect the absence of small mammal or reptile captures. Two A. maculatum were collected, and we report new distribution records in Tennessee for this species. Despite unequal sampling among ecoregions, the large numbers of D. variabilis and A. americanum from multiple host species suggest their widespread distribution throughout the state. These species of ticks can transmit multiple pathogens, including spotted fever group rickettsiae and ehrlichiae.

Vector competence of rural and urban strains of Aedes (Stegomyia) albopictus (Diptera: Culicidae) from São Paulo State, Brazil for IC, ID, and IF subtypes of Venezuelan equine encephalitis virus.

Aedes albopictus (Skuse) is an Asiatic mosquito species that has spread and colonized all continents except Antarctica. It has major public health importance because it is a potential vector of several pathogens. The objectives of our study were to analyze the vector competence of urban and rural strains of Ae. albopictus from São Paulo State (Brazil) for Venezuelan equine encephalitis virus (VEE) subtypes IC, ID, and IF, and to evaluate the effect of infection with subtype IC of VEE on mosquito longevity. Both mosquito strains were susceptible to subtypes IC and ID, but the infection rate for subtype IF was low. Infection and transmission rates of Ae. albopictus for subtype IC were similar to those reported for Ochlerotatus taeniorhynchus (Wiedemann). The high infection, dissemination, and transmission rates for subtype ID reported for Oc. fulvus (Wiedemann) and Culex (Melanoconion) spp. are comparable with those found in this study. We found significant differences in the susceptibility to subtype IC between rural and urban populations of São Paulo. Significant survival rate differences were observed between uninfected and infected mosquitoes, but there were no differences in survival between rural and urban mosquito strains.

Allpahuayo virus: a newly recognized arenavirus (arenaviridae) from arboreal rice rats (oecomys bicolor and oecomys paricola) in northeastern peru.

Allpahuayo virus was initially isolated from arboreal rice rats (Oecomys bicolor and Oecomys paricola) collected during 1997 at the Allpahuayo Biological Station in northeastern Peru. Serological and genetic studies identified the virus as a new member of the Tacaribe complex of the genus Arenavirus. The small (S) segment of the Allpahuayo virus prototype strain CLHP-2098 (Accession No. AY012686) was sequenced, as well as that of sympatric isolate CLHP-2472 (Accession No. AY012687), from the same rodent species. The S segment was 3382 bases in length and phylogenetic analysis indicated that Allpahuayo is a sister virus to Pichinde in clade A. Two ambisense, nonoverlapping reading frames were identified, which result in two predicted gene products, a glycoprotein precursor (GPC) and a nucleocapsid protein (NP). A predicted stable single hairpin secondary structure was identified in the intergenic region between GPC and NP. Details of the genetic organization of Allpahuayo virus are discussed.

Meconial peritrophic membranes and the fate of midgut bacteria during mosquito (Diptera: Culicidae) metamorphosis.

The location of midgut bacteria relative to meconial peritrophic membranes (MPMs) and changes in bacterial numbers during midgut metamorphosis were studied in Anopheles punctipennis (Say), Culex pipiens (L.), and Aedes aegypti (L.) pupae and newly emerged adults. After adult emergence in Aedes, Anopheles, and most Culex, there were few to no bacteria in the midgut. In most newly emerged adult mosquitoes, few bacteria were found in either the lumen or within the MPMs/meconia. In a few Culex specimens, high numbers of bacteria were found in the MPMs/meconia and low numbers in the lumen. In all three species bacterial counts were high in fourth instars, decreased after final larval defecation, increased in young pupae, and increased further in old pupae. A very effective gut sterilization mechanism is operating during mosquito metamorphosis and adult emergence. This mechanism appears to involve the sequestration of remaining larval gut bacteria within the confines of the meconium and one or two MPMs and the possible bactericidal effect of the exuvial (molting) fluid, which is ingested during the process of adult emergence.

Genetic diversity and relationships among Venezuelan equine encephalitis virus field isolates from Colombia and Venezuela.

During field studies of enzootic Venezuelan equine encephalitis (VEE) viruses associated with epizootic emergence, a large number of virus isolates were made in sylvatic foci of Venezuela and Colombia. To rapidly characterize these isolates, antigenic subtypes were determined by means of immunofluorescence and by single-strand conformational polymorphism (SSCP) analysis by use of an 856-bp fragment from the P62 gene, which we used to distinguish genetic variants. Representative isolates were sequenced to assess the sensitivity of SSCP to detect genetic differences. The SSCP analysis distinguished isolates differing by as little as 1 nucleotide; overall, differences of > or = 1 nucleotide were recognized 89% of the time, and the sensitivity to distinguish strains that differed by only 1 or 4 nucleotides was 17 and 57%, respectively. Phylogenetic analyses of representative sequences showed that all recent isolates from the Catatumbo region of western Venezuela and the middle Magdalena Valley of Colombia were closely related to epizootic subtype IAB and IC strains; strains from Yaracuy and Miranda States were more distantly related. Cocirculation of the same virus genotype in both Colombian and Venezuelan foci indicated that these viruses are readily transported between enzootic regions separated by > 300 km. The SSCP analysis appears to be a simple, fast, and relatively efficient method of screening VEE virus isolates to identify meaningful genetic variants.

The occurrence and fate of the meconium and meconial peritrophic membranes in pupal and adult mosquitoes (Diptera: Culicidae).

In mosquitoes, in addition to larval and adult peritrophic membranes (PMs), a PM (meconial peritrophic membrane or MPM1) forms in the pupa around the meconium, the sloughed, degenerating larval midgut epithelium. Often, a second membrane (MPM2) forms in temporal proximity to adult emergence. Differences in the occurrence, persistence, and timing of disappearance of the meconium/MPMs and gas were studied by dissecting the midgut contents from pupae of known ages postpupation and from adults of known ages postemergence. MPM1 was found in all Anopheles and Culex studied and nearly all Culiseta. The occurrence of MPM1 varied in the Aedes species. In one series of Aedes aegypti (L.) dissections, no fully formed MPM2 was found in any specimens. The occurrence of MPM2 appeared to be associated with adult emergence and varied among and within the seven species studied. It typically was seen in recently emerged adults but was observed occasionally in old pupae. Much of our data supports the idea that MPM2 formation is stimulated by midgut epithelium distention.

Effect of eastern equine encephalomyelitis virus on the survival of Aedes albopictus, Anopheles quadrimaculatus, and Coquillettidia perturbans (Diptera: Culicidae).

The effect of eastern equine encephalomyelitis (EEE) virus on the survivorship of Aedes albopictus (Skuse), Anopheles quadrimaculatus Say, and Coquillettidia perturbans (Walker) was determined experimentally. Female mosquitoes were allowed to feed on EEE viremic chicks, and survival rates were compared for infected and uninfected mosquitoes. Additionally, the survival of female Cq. perturbans and An. quadrimaculatus intrathoracically (i.t.) inoculated with EEE was compared with controls receiving diluent inoculations. Infection with EEE significantly reduced survival in Cq. perturbans compared with uninfected individuals in per os infection experiments. I.t. infections of Cq. perturbans did not reduce survival when compared with diluent inoculated groups. In contrast, infection with EEE did not affect the survival of Ae. albopictus after per os infection or An. quadrimaculatus after either i.t. or per os infections.

Application of geographic information technology in determining risk of eastern equine encephalomyelitis virus transmission.

Geographic information system (GIS) technology and remote sensing were used to identify landscape features determining risk of eastern equine encephalomyelitis virus (EEE) transmission as defined by the abundance of Culiseta melanura (the enzootic vector) and 6 putative epidemic-epizootic vectors in Massachusetts. Landsat Thematic Mapper data combined with aerial videography data were used to generate a map of landscape elements at epidemic-epizootic foci in southeastern Massachusetts. Geographic information system technology was used to determine the proportion of landscape elements surrounding 15 human and horse case sites where abundance data were collected for Culiseta melanura, Aedes canadensis, Aedes vexans, Culex salinarius, Coquillettidia perturbans, Anopheles quadrimaculatus, and Anopheles punctipennis. The relationships between vector abundance and landscape proportions were analyzed using stepwise linear regression. Stepwise regression indicated wetlands as the most important major class element, which accounted for up to 72.5% of the observed variation in the host-seeking populations of Ae. canadensis, Ae. vexans, and Cs. melanura. Moreover, stepwise linear regression demonstrated deciduous wetlands to be the specific wetland category contributing to the major class models. This approach of utilizing GIS technology and remote sensing in combination with street mapping can be employed to identify deciduous wetlands in neighborhoods at risk for EEE transmission and to plan more efficient schedules of pesticide applications targeting adults.

Toward the incrimination of epidemic vectors of eastern equine encephalomyelitis virus in Massachusetts: abundance of mosquito populations at epidemic foci.

Putative epidemic/epizootic eastern equine encephalomyelitis virus (EEE) vector populations were compared at 15 recent (1982-90) human and horse case sites in Bristol and Plymouth counties in southeastern Massachusetts. Carbon dioxide-baited American Biophysics Corporation light traps were used for trapping adult mosquitoes to estimate biting risk in these foci of known transmission. Population data suggest that Coquillettidia perturbans, Aedes canadensis, and Culex salinarius are more likely vectors of EEE in Massachusetts than Aedes vexans, Anopheles punctipennis, and Anopheles quadrimaculatus.