Pre-imaginal development times of <i>w</i>MelPop-CLA infected PGYP1 and tetracycline-cured PGYP1.tet <i>Ae. aegypti</i> strains.

<p><i>^a</i>Number of replicates for each strain denoted in parentheses.</p><p>*Significantly different development time (<i>P</i><0.05, MWU test).</p

Age-associated decline in fecundity of PGYP1 and PGYP1.tet strains.

<p>(A) Average number of eggs oviposited per female ± SE. (B) Average number of larvae produced per female ± SE, and (C) Proportion of sampled females that did not oviposit. Females were assayed over successive gonotrophic cycles until death (<i>n</i> = 48 females per time-point). As death occurred over time, samples sizes decreased below 48 females in cycle 7 for PGYP1 females (<i>n</i> = 22), and in cycles 13–16 for PGYP1.tet females (<i>n</i> = 22, 12, 5, and 5 respectively).</p

Viability of quiescent embryos from PGYP1 and PGYP1.tet strains over time at different temperatures.

<p>After embryonic maturation (120 h post oviposition), eggs were stored at either: (A) 25°C and (B) 18°C, with 85% relative humidity. Average proportion of eggs hatching (<i>n</i> = 20 oviposition papers per time point) and standard error bars are shown.</p

Fly lines and <i>Wolbachia</i> strains.

<p>Fly lines and <i>Wolbachia</i> strains.</p

The effect of different <i>Wolbachia</i> strains on the accumulation of DCV in <i>D. simulans</i>.

<p>Graphs show accumulation of infectious DCV in flies with (grey bar) or without (white bar) <i>w</i>Au (<b>A</b>), <i>w</i>Ri (<b>B</b>), <i>w</i>Ha (<b>C</b>), and <i>w</i>No (<b>D</b>). Bars represent means from two replicates with SEM shown, and * indicates a significant difference between the means of day 2 samples (p<0.05, unpaired t test).</p

Relative-density of <i>Wolbachia</i> strains in <i>D. simulans</i>.

<p>For each fly line the graph shows the relative abundance of <i>Wolbachia</i> to host genomic DNA estimated using quantitative PCR. Bars represent the mean of 10 replicates and error bars are SEM.</p

The effect of different <i>Wolbachia</i> strains on the accumulation of FHV in <i>D. simulans.</i>

<p>Graphs show survival of flies infected by <i>w</i>Au (<b>A</b>), <i>w</i>Ri (<b>B</b>), <i>w</i>Ha (<b>C</b>), and <i>w</i>No (<b>D</b>) challenged with FHV (black line) or mock infected (grey line). <i>Wolbachia</i> infected (circle and plus sign) and uninfected (triangle and cross) flies. Error bars represent SEM calculated from three replicates. The survival of FHV infected flies with and without <i>Wolbachia</i> is significantly different for <i>w</i>Au and <i>w</i>Ri (p<0.0001, log rank test on Kaplan-Meier curves). For each fly line a similar result was recorded in a replicate experiment.</p

<i>Wolbachia</i> strain <i>w</i>Mel provides antiviral protection in <i>D. simulans</i>.

<p>(<b>A</b>) Graph shows survival of flies infected with DCV (black line) or mock infected (grey line). <i>w</i>Mel-infected (circle and plus sign) or uninfected (triangle and cross) flies. The survival of DCV infected flies with and without <i>Wolbachia</i> is significantly different (p<0.0001). Error bars represent SEM calculated from three replicate vials. This is a representative experiment which was repeated twice more with similar results. (<b>B</b>) Graph showing accumulation of infectious DCV in <i>w</i>Mel infected (grey bars) or uninfected (white bar) flies. Bars represent means from two replicates with SEM shown, and * indicates a significant difference between the means of day 2 samples (p<0.05, unpaired t test).</p

Antiviral protection of different <i>Wolbachia</i> strains in <i>D. simulans</i>.

<p>Graphs show survival of flies infected by <i>w</i>Au (<b>A</b>), <i>w</i>Ri (<b>B</b>), <i>w</i>Ha (<b>C</b>), and <i>w</i>No (<b>D</b>) challenged with DCV (black line) or mock infected (grey line). Flies with <i>Wolbachia</i> (circle and plus sign) and without <i>Wolbachia</i> (triangle and cross). Error bars represent SEM calculated from three replicates. The survival of DCV infected flies with and without <i>Wolbachia</i> is significantly different for <i>w</i>Au (p<0.0001), <i>w</i>Ri (p<0.0001), and <i>w</i>Ha (p<0.01), using log rank test on Kaplan-Meier curves. Experiments were replicated on at least two additional independent cohorts of flies, and the results for all respective replicates of experiments shown in panel A, B and D were similar, however the replicates for panel C varied (see <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000656#s2" target="_blank">Results</a>).</p

Survival curves of <i>Wolbachia</i>-infected (circle) and <i>Wolbachia</i>-uninfected (square) Drosophila infected with pathogenic bacteria (solid) or mock infected with PBS (open).

<p>(A&amp;E) <i>E. carotovora</i>, (B&amp;F) <i>B. cepacia</i>, (C&amp;G) <i>S. typhimurium</i> and (D&amp;H) <i>M. marinum</i>. Error bars are SEM calculated from the three replicates. * P-value&lt;0.05, ** P-value&lt;0.01, *** P-value&lt;0.001 denote differences in survival between <i>Wolbachia</i> infected and uninfected lines by Log-rank statistics (<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002362#pntd.0002362.s001" target="_blank">Table S1A</a>).</p