Fig 5 -

MTT assay of cloned cell lines from A10/BT (A) and A21/AT1 (B) isolates. Numbers on the x axis (-1, 0, 1, 2, 3 and 4) indicates drug concentrations used to treat cell lines at doses of 0, 0.8, 7, 125, 1000 and 1500 ng/mL, respectively.</p

Alignments of putative Q_o1 and Q_o2 domains of <i>Cyto b</i>.

Predicted amino acid sequences of Qo1 and Qo2 domains encoded by the Cyto b gene from the published T. annulata / C9 genome sequence and different T. annulata isolates with treatment failure history. Putative Qo1 and Qo2 domains are located between 116–144 and 238–273 amino acids of the protein around the ubiquinone binding site. Predicted amino acid substitutions associated with buparvaquone resistance are marked with ◊.</p

Number of alleles detected by nine micro- and mini-satellite markers before and after buparvaquone treatments.

Number of alleles detected by nine micro- and mini-satellite markers before and after buparvaquone treatments.</p

Geographical distribution of the parasite material used in the present study.

The geographical distribution of the provinces where the parasite material used in this study was obtained. The map was prepared using the USGS National Map Viewer (public domain): http://viewer.nationalmap.gov/viewer/ with some modification. (TIF)</p

Fig 4 -

Alignments of TaPIN1 nucleotide (A) and predicted amino acid (B) sequences. Sequences derived from a buparvaquone resistant Tunisian isolate and various Turkish T. annulata isolates with treatment failure history were compared for mutations with the T. annulata / C9 reference genome, obtained from PiroplasmDB.org database; the systematic identifier for TaPIN1 is TA18945. Nonsynonymous substitutions detected among alignments were indicated with an asterisk (*) coloured in yellow.</p

Agarose gel electrophoresis of AS-PCR.

Gels showing detection of PCR amplicons of isolates with mutations V135A (A) and P253S (B), respectively. Products amplified using drug sensitive and resistance specific forward primers are given at the top and bottom of each gel, respectively. M, 100 bp molecular size marker (Thermo Scientific Corp.); lanes 1–16, products from template DNA of T. annulata samples; lanes S sensitive control, lanes R resistance control, lane X mixed control. (TIF)</p

IC₅₀ values of A10/BT and A21/AT1 clones with substitutions V135A and P253S.

IC50 values of A10/BT and A21/AT1 clones with substitutions V135A and P253S.</p

MTT assay of <i>T</i>. <i>annulata</i> infected cell lines.

A.T. annulata isolates with low IC50 values (2–3 ng/mL). B. T. annulata isolates with medium IC50 values (3–7 ng/mL). C. T. annulata isolates with IC50 values over (7 ng/mL). Numbers on the x axis (-1, 0, 1, 2, 3 and 4) indicates drug concentrations used to treat cell lines at doses of 0, 0.8, 7, 125, 1000 and 1500 ng/mL, respectively.</p

Summary of mutations detected in <i>Cyto b</i> sequences of <i>T</i>. <i>annulata</i> isolates from Tunisia, Sudan, Iran and Turkey.

(PDF)</p

Fig 2 -

MTT assay of T. annulata infected cell lines obtained from animals A10 (A) and A21 (B) following repeated buparvaquone treatments. Numbers on the x axis (-1, 0, 1, 2, 3 and 4) indicates drug concentrations used to treat cell lines at doses of 0, 0.8, 7, 125, 1000 and 1500 ng/mL, respectively.</p