Tularemia has long been classified as an infection of natural focality/nidality. The agents for such infections survive for GSK621 solubility dmso extended durations, decades or longer, in discrete sites (“”natural foci”") characterized by specific faunal, floral, and physical associations. [16] We have subsequently confirmed, by the use of GIS mapping and VNTR analysis, the natural nidality of F. tularensis tularensis on Martha’s Vineyard. [17] Ultimately, we seek to better understand the factors that

serve as the basis for epizootics as opposed to cryptic maintenance within natural foci. Our hypothesis is rooted in metapopulation ecology [18, 19]: that F. tularensis tularensis exists in multiple small, isolated natural foci, in which genetic drift increases diversity until some adaptive equilibrium PI3K inhibitor is achieved. When local conditions

change, such as increased density of hosts for subadult dog ticks, “”valleys”" between such adaptive peaks are traversed and certain strains escape to mix into other “”peaks”" or establish new ones. Natural selection then operates to homogenize the genetic structure across the metapopulation of natural foci. As a first step in exploring this hypothesis, we examined the population structure of two different sites that are separated by 15 km on the island, a natural focus that has long-term stable transmission and a focus that is PAK5 newly emerging. In particular, we sought to determine whether the force of transmission between the two sites differed, and using VNTR analysis of F. tularensis DNA from host seeking dog ticks, we sought evidence for their genetic isolation. Methods Tick collection Collections were conducted from 2003–2007 monthly from April to August. Questing D. variabilis were OTX015 order obtained by flagging the vegetation. Additional ticks were obtained by removing them from skunks and raccoons (< 6%

of the ticks included in the study) as previously described. [13] Sampling was done from two field sites on opposite sides of the island, near Squibnocket and Katama (see Figure 1). The Squibnocket site is what we believe to comprise a longstanding elementary focus. In contrast, Katama is a site where D. variabilis is exceedingly dense but where F. tularensis tularensis appears to be rare. Both sites are similar in physiography, with coastal grassland and beach scrub proximal to large brackish water ponds. Both are undeveloped areas of glacial outwash plains with scrubby barrier beach habitat, although the Katama site experiences intensive seasonal use by people for beach access. Figure 1 Collection sites on Martha’s Vineyard. PCR A drop of hemolymph was obtained from each tick by cutting the front foreleg. This was placed in a tube containing 50 ul PBS. Ticks were processed in pools of 6. Ticks were held at 15°C in individual tubes during screening.