*4.2. Parasites*

The standard strain of the *Leishmania major* parasite is MHOM/TN/94/GLC94 (GLC94), which was isolated in a human lesion of zoonotic cutaneous leishmaniasis (ZCL). The most virulent strain of a parasite is the *L. major* zymodeme MON25 (*L. major*-MON25) [31–33]. Promastigotes were collected in a logarithmic-phase culture, counted in a fixed volume, and then incubated once more at 26 ◦C, as previously mentioned. After five days, the stationary phase was attained, and the parasite concentration value was equal to 4 × 10<sup>7</sup> parasites/milliliter. Then, using the density gradient centrifugation method, metacyclic promastigotes were isolated, cleaned, and administered to BALB/c mice for inoculation (2 × 106).
