In this research, community level spatial models were developed for determining mosquito abundance and environmental factors that could aid in the risk prediction of West Nile virus (WNv) outbreaks. Adult Culex pipiens and Culex restuan mosquitoes and multiple habitat covariates were collected from nine sites within Cook County, Illinois, USA, to provide spatio-temporal information on the abundance of WNv vectors from 2002 to 2005. Regression analyses of the sampled covariates revealed that the adult Culex population was positively associated with temperature throughout the sampling frame. The model output also indicated that precipitation was negatively associated to mosquito abundance in 2002, 2003 and 2005 (P <0.05), but positively associated in 2004 (P <0.05). A land use land cover classification, based on QuickBird visible and near infra-red data, acquired at 0.61 m resolution, was used to investigate possible associations between geographical features and the abundance of sampled Culex oviposition surveillance sites. A maximum likelihood unsupervised classification in ArcInfo 9.2(R) revealed that the highest overall mosquito abundance was found in sites having a low-to-moderate range of built environment (40%) and high forest composition. A set of propagation equations were then designed to model the calibration uncertainties, which revealed that normalized difference vegetation index (NDVI), and two NDVI variants, were informative markers for the sampled mosquito data. Spatial dependence of the covariates of Cx. restuans and Cx. pipiens oviposition sites were indexed using semivariograms, which suggested that all main effects of the explanatory variables were statistically significant in the model. Additionally, a multispectral classification and digital elevation model-based geographical information system method were able to evaluate stream flow direction and accumulation for identification of terrain covariates associated with the sampled habitat data. These results demonstrate that remotely sensed operational indices can be used to identify parameters associated with field-sampled Cx. pipiens and Cx. restuans aquatic habitats.

Two promoters from the human filarial parasite Brugia malayi have been mapped in detail. The essential domains of both promoters lacked canonical eukaryotic core promoter motifs. However, the largest contiguous essential domain in both promoters flanked and included the splice leader addition site. These findings suggested that the region flanking the trans-splicing addition site might represent a conserved core domain in B. malayi promoters. To test this hypothesis, the putative promoters of 12 trans-spliced genes encoding ribosomal protein homologues from B. malayi were isolated and tested for activity in a B. malayi transient transfection system. Of the 12 domains examined, 11 produced detectable reporter gene activity. Mutant constructs of the six most active promoters were prepared in which the spliced leader acceptor site and the10 nt upstream and downstream of the site were deleted. All deletion constructs exhibited >90% reduction in reporter gene activity relative to their respective wild type sequences. A conserved pyrimidine-rich tract was located directly upstream from the spliced leader splice acceptor site which contained a conserved T residue located at position -3. Mutation of the entire polypyrimidine tract or the conserved T individually resulted in the loss of over 90% of reporter gene activity. In contrast, mutation of the splice acceptor site did not significantly reduce promoter activity. These data suggest that the region surrounding the splice acceptor site in the ribosomal promoters represents a conserved essential domain which functions independently of splice leader addition.

Traditional environmental management programmes require extensive coverage of larval habitats to reduce drastically the emergence of adult mosquitoes. Recent studies have highlighted the impact of reduced availability of aquatic habitats on mosquito foraging for hosts and oviposition sites. In this study, we developed an agent-based model to track the status and movement of mosquitoes individually. Mosquito foraging was represented as a two-stage process: random flight when the resource was not within the mosquito's perception range and directional flight to the resource when it was detected. Three scenarios of targeted source reduction were devised to eliminate all aquatic habitats within certain distances of human habitations. For comparison, three non-targeted source reductions randomly eliminated the same numbers of aquatic habitats as their corresponding targeted scenarios. Our results show that the elimination of habitats within 100m, 200m and 300m of surrounding houses resulted in 13%, 91% and 94% reductions in malaria incidence, respectively; compared with -3%, 19% and 44%, respectively, for the corresponding conventional interventions. These findings indicate that source reduction might not require coverage of extensive areas, as previously thought, and that the distance to human habitations can be used for habitat targeting.

Reptiles and amphibians constitute a significant portion of vertebrate biomass in terrestrial ecosystems and may be important arbovirus reservoirs. To investigate mosquito preference for ectothermic hosts, feeding indices were calculated from data collected in Tuskegee National Forest, Alabama, USA. Four mosquito species fed upon ectothermic hosts, with Culex peccator and Cx. territans feeding primarily upon ectotherms. These two species appeared to target distinct species with little overlap in host choice. Culex peccator was a generalist in its feeding patterns within ectotherms, and Cx. territans appeared to be a more specialized feeder. Six of eleven ectotherm species fed upon by Cx. territans were fed upon more often than predicted based upon abundance. Spring peepers were highly preferred over other host species by Cx. territans. Blood meals taken from each host species varied temporally, with some hosts being targeted fairly evenly throughout the season and others being fed upon in seasonal peaks.

The identification of the members of the Culex pipiens L. complex in arbovirus surveillance programs relies heavily on the use of morphology. In this work, we studied Cx. pipiens complex male mosquitoes collected from nine different locations, from northern, southern, and the hybrid zone sites in North America; Cairo, Egypt; and Nairobi, Kenya. Specimens were identified using DV/D ratio and also using amplification of the acetylcholinesterase (ACE.2) gene by both conventional and real-time polymerase chain reaction, and examination of the CQ11 locus. Consistent with previous findings, the morphological and molecular identifications did not always agree, particularly in regions of extensive introgression. There was an increased frequency of hybrid forms in late summer and early fall in Champaign Co., IL, that is north of the previously described Cx. pipiens complex hybrid zone. This represents an expansion of the North American hybrid zone. The biological and epidemiological relevance of the high degree of introgression and the late season increase in the proportion of intermediate forms is discussed.

Unravelling gene regulatory mechanisms in human filarial parasites will require an understanding of their basic promoter structure. Only a single promoter from a human filarial parasite has been characterised in detail, the 70kDa heat shock promoter of Brugia malayi (BmHSP70). This promoter was found to lack features found in a typical eukaryotic promoter. To determine if this was unique to the BmHSP70 promoter, a detailed analysis was undertaken of the promoter for the B. malayi small subunit 12kDa ribosomal protein (BmRPS12) gene. The BmRPS12 promoter contained a unique tandem repeat structure. Deletion of these repeats resulted in the loss of 80% of promoter activity. Block replacement mutagenesis identified five regions outside the repeat which were essential for promoter activity. No predicted binding sites for proteins that normally associate with the typical eukaryotic core promoter domains were found in the essential domains or the repeat region. However, the repeat region contained many putative binding sites for GATAA transcription factor family proteins. Of 20 upstream domains of other ribosomal protein genes, one contained a repeat structure similar to that found in the BmRPS12 promoter, and the majority encoded putative GATAA transcription factor binding sites. This study demonstrates that the BmRPS12 promoter, like the BmHSP70 promoter, is distinct from a typical eukaryotic promoter.

Interest in environmental management of mosquito larval habitats has been rekindled due to deterioration of malaria in tropical Africa. Environmental management programs were typically implemented as "all-out" campaigns by treating all potential breeding habitats. In contrast, targeted environmental management is based on a sound understanding of the heterogeneity in mosquito productivity. However, deficiencies in field methodology for measuring productivity hamper our progress in understanding of mosquito productivity. To address these issues, we develop a framework of habitat-based interventions by adoption of a landscape approach to elucidate mechanisms underlying mosquito productivity. The importance of vigorously quantitative estimation of the productivity is highlighted. Spatial models are proposed to examine the interrelationship between mosquito productivity and oviposition of gravid mosquitoes. In our view, environmental management approaches must take into account variability in productivity, in efforts to improve feasibility, cost-effectiveness, and sustainability of such approaches, particularly when implemented along with other malaria control measures.

The source of bloodmeals in 2,082 blood-fed mosquitoes collected from February 2002 through December 2003 in Memphis and surrounding areas of Shelby County, Tennessee were determined. Members of the genus Culex and Anopheles quadrimaculatus predominated in the collections. Members of the Cx. pipiens complex and Cx. restuans were found to feed predominately upon avian hosts, though mammalian hosts made up a substantial proportion of the bloodmeals in these species. No significant difference was seen in the host class of bloodmeals in mosquitoes identified as Cx. pipiens pipiens, Cx. p. quinquefasciatus, or hybrids between these two taxa. Anopheles quadrimaculatus and Cx. erraticus fed primarily upon mammalian hosts. Three avian species (the American Robin, the Common Grackle, and the Northern Cardinal) made up the majority of avian-derived bloodmeals, with the American Robin representing the most frequently fed upon avian host. An analysis of these host feeding data using a modification of a transmission model for Eastern Equine encephalitis virus suggested that the American Robin and Common Grackle represented the most important reservoir hosts for West Nile virus. A temporal analysis of the feeding patterns of the dominant Culex species did not support a shift in feeding behavior away from robins to mammals late in the summer. However, a significant degree of temporal variation was noted in the proportion of robin-derived bloodmeals when the data were analyzed by semi-monthly periods throughout the summers of 2002 and 2003. This pattern was consistent with the hypothesis that the mosquitoes were preferentially feeding upon nesting birds.

In North America, West Nile and St. Louis encephalitis viruses have been detected in a wide range of vector species, but the majority of isolations continue to be from pools of mixed mosquitoes in the Culex subgenus Culex. Unfortunately, the morphologic identification of these important disease vectors is often difficult, particularly in regions of sympatry. We developed a sensitive real-time TaqMan polymerase chain reaction assay that allows reliable identification of Culex mosquitoes including Culex pipiens pipiens, Cx. p. quinquefasciatus, Cx. restuans, Cx. salinarius, Cx. nigripalpus, and Cx. tarsalis. Primers and fluorogenic probes specific to each species were designed based on sequences of the acetylcholinesterase gene (Ace2). Both immature and adult mosquitoes were successfully identified as individuals and as mixed species pools. This identification technique provides the basis for a rapid, sensitive, and high-throughput method for expounding the species-specific contribution of vectors to various phases of arbovirus transmission.