Infection prevalence and viral load throughout EHDV-2 infection within the midge.

<p>Infection prevalence and viral load throughout EHDV-2 infection within the midge.</p

IHC staining of EHDV-2 infected tissues throughout <i>C</i>. <i>sonorensis</i>.

<p>Representative images were taken from orally infected midges at 7–10 dpf. (<b>A</b>) The internal anatomy of <i>C</i>. <i>sonorensis</i> revealed by a hematoxylin-stained sagittal section of the whole midge. No staining for EHDV-2 is provided in this section. Boxes indicate locations of tissues positive for EHDV-2 by IHC staining (red). (<b>B</b>) the neural lamella (dotted line) of the cerebral ganglia (cg); (<b>C</b>) salivary gland (sg), fat body (fb), trachea (t); (<b>D</b>) posterior midgut, cuticle (c); (<b>E</b>) ovarian sheath (os), ovariole (o); (<b>F</b>) Johnston’s organ (antenna); (<b>G</b>) eye containing the ommatidia (om) and optical ganglia (og); (<b>H</b>) fat body (fb); and (<b>I</b>) spermatheca, which contains the spermathecal epithelia (se), reservoir lumen (rl), and sperm (arrow). All sections were counterstained with hematoxylin (blue) to reveal overall tissue structure. Scale bars: (A) = 200 μm, (B-D and F-I) = 25 μm, and (E) = 50 μm.</p

Whole body viral titer over the course of EHDV-2 infection within <i>C</i>. <i>sonorensis</i>.

<p>Graph depicts the viral load per midge over the infection time course. Total VPs (black), were estimated by calculating genome equivalents (GE) by qPCR, and infectious VPs (red with 10^1.39 PFU/midge limit of detection: dotted line) were calculated by plaque assay. All samples are represented as median and interquartile range. Corresponding raw values are presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0188865#pone.0188865.t001" target="_blank">Table 1</a>.</p

EHDV-2 positive staining pattern in <i>C</i>. <i>sonorensis</i> by IHC over the course of infection.

<p>Increasing intensity of IHC staining (red) showing progression of EHDV-2 infection in tissues over time, with representative images taken at 5 dpf (<b>B, E, H</b>, and <b>K</b>) and 8 dpf (<b>C, F, I</b>, and <b>L</b>), compared to negative control tissues (<b>A, D, G</b>, and <b>J</b>). This increase was observed in all infected tissues: (<b>A-C</b>) the midgut [midgut epithelium (e), fat body (fb), cuticle (c)]; (<b>D-F</b>) salivary glands (sg), [trachea (t)]; (<b>G-I</b>) fat body (fb); and (<b>J-L</b>) eye containing the ommatidia (om) and optic ganglia (og) [cerebral ganglia (cg)]. All sections were counterstained with hematoxylin (blue) to reveal overall tissue structure. Scale bars = 25 μm.</p

Bar graphs showing by-species patterns of change in RVFV seropositivity before, during, and after the 2006–2007 RVF epizootic.

<p>Bar graphs showing by-species patterns of change in RVFV seropositivity before, during, and after the 2006–2007 RVF epizootic.</p

RVF risk quantified in a 75 km radius from each sample location.

<p>RVF risk quantified in a 75 km radius from each sample location.</p

Monthly predicted RVF risk assessment map overlaid with serological results collected after the 2006–2007 RVF outbreak; A) Marsabit, Mandera, and Wajir in January and February 2008, B) Marsabit, Mandera, and Wajir in August and September 2008, C) Ijara, Klegdela, Marsabit, Meru, Tana, and Tsavo in October and November 2008.

<p>See Fig. 4 for descriptive legend.</p

Map of Kenya showing 2000–2009 serology sample locations and relevant Kenya conservation areas.

<p>Map of Kenya showing 2000–2009 serology sample locations and relevant Kenya conservation areas.</p

Bar graphs showing by-location patterns of change in RVFV seropositivity before, during, and after the 2006–2007 RVF epizootic.

<p>Bar graphs showing by-location patterns of change in RVFV seropositivity before, during, and after the 2006–2007 RVF epizootic.</p

Wildlife sera collected in different locations during pre-epizootic, epizootic, and post epizootic RVF outbreak in Kenya.

<p>The results of the analysis of the RVFV antibodies in the samples by recombinant N inhibition ELISA are indicated.</p