*4.6. Haematological Procedures*

The haematological profile consisted of total white (WBC) and red (RBC) blood cells counts. To determine WBC and RBC concentration, whole blood was diluted 1/20 and 1/200, respectively, in HBSS with heparin (30 U mL−1) and cell counts were done in a Neubauer chamber. Blood smears were prepared from peripheral blood, air-dried and stained with Wright's stain (Haemacolor; Merck, Darmstadt, Germany), after fixation for 1 min with formol–ethanol (10% formaldehyde in ethanol). Neutrophils were labelled by detecting peroxidase activity revealed by Antonow's technique described in Afonso et al. [56]. The slides were examined under oil immersion (1000×), and at least 200 leucocytes were counted and classified as thrombocytes, lymphocytes, monocytes and neutrophils. The relative percentage and absolute value (×104 mL<sup>−</sup>1) of each cell type was calculated.
