Fig 1.
Dose-response curves of TC against Babesia and Theileria parasites in vitro.
The curves show the relative fluorescence units of B. bovis, B. bigemina, B. divergens, B. caballi, and T. equi treated with increasing concentrations of TC. The results were determined via fluorescence assay after 96 h of incubation in three separate trials. The values obtained from three separate trials were used to determine the IC50 values using the non-linear regression (curve fit analysis) in GraphPad Prism software (GraphPad Software Inc., USA).
Fig 2.
Dose-response curves of CH against Babesia and Theileria parasites in vitro.
The curves show the relative fluorescence units of B. bovis, B. bigemina, B. divergens, B. caballi, and T. equi treated with increasing concentrations of CH. The results were determined via fluorescence assay after 96 h of incubation in three separate trials. The values obtained from three separate trials were used to determine the IC50 values using the non-linear regression (curve fit analysis) in GraphPad Prism software (GraphPad Software Inc., USA).
Table 1.
The IC50 and selectivity index of TC and CH.
Table 2.
The viability of Babesia and Theileria parasites treated with TC and CH.
Fig 3.
The morphological changes observed in TC- and CH-treated B. bovis.
The arrows show TC- and CH-treated B. bovis parasites. The micrographs, C, were taken from the untreated wells. TC-a and CH-a were taken from treated wells at 24 h, while TC-b and CH-b were taken at 72 h.
Fig 4.
The morphological changes observed in TC- and CH-treated B. caballi.
The arrows show TC- and CH-treated B. caballi parasites. The micrographs, C, were taken from the untreated wells. TC-a and CH-a were taken from treated wells at 24 h, while TC-b and CH-b were taken at 72 h.
Table 3.
The effect of TC or CH with DA, AQ, or CF against Babesia and Theileria parasites in vitro.
Fig 5.
The growth inhibition of TC on B. microti in vivo.
The graph shows the inhibitory effects of TC, DA, AQ, and CF treatments as compared with the untreated group. The values plotted indicate the mean ± standard deviation for two separate experiments. The asterisks (*) indicate statistical significance (p < 0.05) based on one-way ANOVA Tukey's test using GraphPad Prism version 5.0 for Windows (GraphPad Software Inc., San Diego, CA, USA). The arrow indicates 5 consecutive days of treatment. Parasitemia was calculated by counting infected RBCs among 2,000 RBCs using Giemsa-stained thin blood smears.
Fig 6.
The growth inhibition of TC combinations on B. microti in vivo.
The graph shows the inhibitory effects of DA, AQ, and CF combined with TC treatments as compared with the untreated group. The values plotted indicate the mean ± standard deviation for two separate experiments. The asterisks (*) indicate statistical significance (p < 0.05) based on one-way ANOVA Tukey's test using GraphPad Prism version 5.0 for Windows (GraphPad Software Inc., San Diego, CA, USA). The arrow indicates 5 consecutive days of treatment. Parasitemia was calculated by counting infected RBCs among 2,000 RBCs using Giemsa-stained thin blood smears.
Fig 7.
Molecular detection of parasite DNA in the treated groups.
The image shows the molecular detection of parasites in the treated groups. The double distilled water (DDW) is for the untreated- uninfected group which used as a negative control, PC is for the untreated- infected group which used as the positive control, and M is for the marker. The arrow shows the expected band length of 154 bp for positive cases of B. microti.
Fig 8.
The changes in blood parameters in treated and untreated mice in vivo.
The graphs show changes in the number of red blood cells (RBCs) (A), hemoglobin concentration (HGB) (B), and hematocrit percentage (HCT) (C) in different groups of treated mice as compared with untreated mice. The values plotted are the mean ± standard deviation for two separate trials. Each group contained five mice. The asterisks (*) indicate statistical significance (p < 0.05) based on one-way ANOVA Tukey's test using GraphPad Prism version 5.0 for Windows (GraphPad Software Inc., San Diego, CA, USA).