Frequency of household interview responses.

<p>*We were informed during Interview #6 of 23 that deworming of pigs was a universal practice in the village. Thus, we began explicitly asking this question in every subsequent household interview, totaling 18 responses.</p><p>Frequency of household interview responses.</p

Map of the study area and location of the six villages in Danzhai, Guizhou, China.

<p>Map of the study area and location of the six villages in Danzhai, Guizhou, China.</p

Algal Uptake of Hydrophobic and Hydrophilic Dissolved Organic Nitrogen in Effluent from Biological Nutrient Removal Municipal Wastewater Treatment Systems

Dissolved organic nitrogen (DON) accounts for a large fraction of the total nitrogen discharged to surface waters by municipal wastewater treatment plants designed for biological nutrient removal (BNR). Previous research indicates that some but not all of the DON in wastewater effluent is available to bacteria and algae over time scales that are relevant to rivers and estuaries. To separate bioavailable DON from nitrate and less reactive DON species, an XAD-8 resin coupled with an anion exchange treatment was employed prior to chemical analysis and algal bioassays. Analysis of effluent samples from a range of municipal BNR plants (total DON concentrations ranging from 0.7 to 1.8 mg N/L) employing a range of technologies indicated that hydrophilic DON, which typically accounted for approximately 80% of the total DON, stimulated algal growth, whereas hydrophobic DON, which accounted for the remaining DON, remained at nearly constant concentrations and had little or no effect on algal growth during a 14-day incubation period. The hydrophobic DON exhibits characteristics of humic substances, and is likely to persist for long periods in the aquatic environment. The distinct differences between these two classes of DON may provide a basis for considering them separately in water quality models and effluent discharge regulations

Assay Concordance plot.

<p>Bivariate scatter correlation plot between vaccinia virus infectivity measured by high-content analysis based infectivity assay (HC-IA) (Y-axis) and the plaque assay (X-axis) presented in log 10 scale. Potencies of thirteen stocks of vaccinia virus were measured in plaque assay and HC-IA. Results were analyzed using GraphPad Prizm software showing significant correlation between image-based and standard plaque counting assays (Pearson r—0.95, P value—0.0001, R²—0.9, slope 0.83). Dashed line represents the ideal linear curve with slope of 1, while solid line is an observed concordance curve.</p

IC₅₀ infectivity values for tested Orthopoxviruses.

<p>IC₅₀ infectivity values for tested Orthopoxviruses.</p

Bivariate scatter correlation plot between two imaging assays.

<p>New Orthopoxvirus neutralization assay (HC-OVNA), Y-axis, and GFP-expressing vaccinia virus (HCS-GFP), X-axis, presented in log 10 scale. Correlation between two assays was evaluated using 49 human serum samples from smallpox vaccine study (CDC IRB protocol 3349). Mean EC₅₀ values from neutralization curves from HC-OVNA and HCS-GFP assays were plotted against each other (log scale) and analyzed using GraphPad Prizm software, showing significant correlation between two assays (Pearson r—0.9, P value—0.0001, R2–0.8).</p

Average immune response for different vaccinee categories.

<p>Neutralization responses for each vaccinee category were grouped according to blood draw days. Calculated average and standard deviation values were based on the results from several vaccinee per each category.</p

Assay reproducibility.

<p>Z-factor experiment was performed to evaluate assay reproducibility, where 20% of a plate had wells infected with vaccinia virus at MOI 0.250, and the rest of the plate had wells that were infected (MOI 0.250) and neutralized with pooled human serum at 1:100 dilution Calculated Z-factor value is 0.63 indicating that the developed assay is robust and reproducible.</p

Variola virus infection in Vero E6 cells at different MOI at 24 hours post infection.

<p>Cell images were collected at 5X magnification with 9 fields per well. Variola virus (Solaimen) was titrated at different MOI concentrations. Wells with uninfected control cells were included on each plate to be used for gating purposes during image analysis. Cells were identified by nuclear staining (pseudo-colored blue) and virus presence was detected with indirect staining (pseudo-colored green). Images show infected cells in one field per well at different MOI after 24 hours infection.</p

Infection dynamics.

<p>Infection dynamics were tested in Vero E6 cells for four different types of Orthopoxviruses. Cells were infected with viral titrations starting from MOI 1 (1.7x10⁴ pfu/well), X-axis, followed by a series of two-fold dilutions down to a lowest multiplicity infection of 3.9x10^-4 for vaccinia virus, and 9.76x10^-4 for variola, cowpox and monkeypox viruses. The Y-axis corresponds to percent of infected cells. Incubation times ranged from 5–28 hours post infection (hpi). NC = Negative control.</p