Use of an in vitro infectivity assay in comparison with histological techniques in the study of Theileria parva sporozoite maturation.

Adult male and female Rhipicephalus appendiculatus ticks infected with Theileria parva (Muguga 3087) were fed on rabbits and the development of infection was monitored daily using light microscopy and an in vitro titration technique able to quantify the infectivity of sporozoite suspensions. The salivary glands stained with methyl green pyronine showed presence of infection in some unfed ticks. The intensity of staining was shown to increase with the number of days the ticks had fed. The in vitro technique, on the other hand, could detect infection only in ticks which had fed for 3-5 days. Feeding of ticks on rabbits for 4 days produced significantly more sporozoites than any other lengths of feeding (P = 0.001). The in vitro assay was also able to demonstrate differences between male and female R. appendiculatus in production of infective sporozoites. Female ticks produced significantly more sporozoites than male ticks (P = 0.002).