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TBS were prepared in duplicate and slides stained with Giemsa. Counting was performed in 500 white blood cells, which corresponded to 25 minutes of observation18. In order to determine the number of parasites/μL, two independent and highly experienced microscopists carried out readings, following the criteria recommended by the Brazilian Ministry of Health, and assuming a standard of 6,000 leucocytes/μL19. TBS parasitaemia was calculated as numbers of parasites/μL:
The same laboratory staff examined the TBS of all three groups.
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