4. 4 Mosquito cycles
Anopheles stephensi mosquitoes were maintained as previously described (Narwal et al. , 2022). To obtain sporozoites, mosquitoes were allowed to feed on infected Swiss mice, and cages were kept in an environmental chamber maintained at 190°C with 80% relative humidity. The development of oocysts was observed on day 14, and salivary gland sporozoites were dissected between days 18-22 postfeeding.