Description of <i>P.papatasi</i> samples collected and screened for gut flora.

#<p>Fly, which did not produce any colony.</p><p><a href="mailto:@Contaminated" target="_blank">@Contaminated</a> sample.</p

<i>Ljunghia pulleinei</i>, female.

<p>A, dorsal idiosoma (with setal notation of some dorsal setae); B, ventral idiosoma. Scale: 100 µm.</p

Distribution of <i>P. papatasi</i> gut bacteria among other hosts.

§<p>this report;</p>¶<p>immature stages only.</p

Distribution of gut flora of adult <i>P. papatasi</i> females.

<p>Distribution of gut flora of adult <i>P. papatasi</i> females.</p

Bacterial clones of sand fly gut flora grown in BHI agar plate showing more than 100 colonies from a single <i>P. papatasi</i> female gut.

<p>Bacterial clones of sand fly gut flora grown in BHI agar plate showing more than 100 colonies from a single <i>P. papatasi</i> female gut.</p

Bayesian 16S tree of gut flora of adult <i>P. papatasi</i> females.

<p>Posterior probabilities are given along internodes. The scale bar denotes substitutions per nucleotide for the branch lengths. Species that have been implicated in inducing oviposition behavior are highlighted in red.</p

2-D Page of <i>Wolbachia</i>-infected and uninfected Aa23 cells.

<p>(A) Identification of proteins unique to <i>Wolbachia-</i>infected Aa23 cells. Approximately 750 ug of protein extract from an Aa23 cell line stably infected with <i>Wolbachia</i> (I) and a parallel cell line cured of a <i>Wolbachia</i> infection (II) were analyzed. Proteins expressed only in the presence of a <i>Wolbachia</i> infection (ID #1–6) are identified. (B) Gel sections showing proteins selected for LC/MS/MS analysis.</p

Analysis of ROS formation in <i>Wolbachia</i> -infected and uninfected Aa23 cells.

<p>(A) Flow cytometric analysis of <i>Wolbachia</i> –infected and uninfected Aa23 cells using the fluorescent ROS marker carboxy-H₂DCFDA. Histograms representative of three replicates are shown. The negative control (shaded) consists of unlabeled cells. Test samples (black lines) include: uninfected Aa23 cells (top panel), uninfected Aa23 cells induced to produce ROS using TBHP (middle panel), and infected Aa23 cells (bottom panel). Carboxy-H₂DCFDA positive cells are represented on each histogram. (B) Microscopic analysis of <i>Wolbachia</i>-infected (I) and uninfected (II) Aa23 cells. Hoechst stain was used to label DNA (left panel). Carboxy-H₂DCFDA was used to label ROS (right panel). Bar, 10 µm.</p

<i>Wolbachia</i> stably infects Aa23 cells and can be cured by antibiotic treatment.

<p>(A) PCR analysis using <i>Wolbachia wsp</i> primers (top) and arthropod 28S primers (bottom) of Aa23 cells treated with 10 ug/ml rifampicin for seven passages. Lane L: molecular ladder. Lane 1: stably infected Aa23 cells. Lanes 2 through 8: cells treated with rifampicin for 1, 2, 3, 4, 5, 6, and 7 passages. Lane 9: negative control. (B) Aa23 cells stably infected with <i>Wolbachia</i> (I) and Aa23 cells cured of <i>Wolbachia</i> using rifampicin (II). Bar, 100 µm.</p

Maximum likelihood phylogram representing phylogenetic relationships of <i>Riesia</i> p-endosymbionts.

<p>Numbers above the node represent divergence dates (in millions of years) whereas numbers below the nodes are bootstrap support values (only numbers greater then 60 are shown). The divergence date calibration point of 9.42–17.38 My is indicated with an asterisk.</p