Natural tick infestations were assessed every 14 days on horses over a 2-year period. Amblyomma cajennense adult ticks were counted individually, without detachment from the horses. Larvae and nymphs of A. cajennense were collected using a rubber scraper that scratched engorged immature ticks from the host. Adult females of Anocentor nitens larger than 4mm length were counted on the horses. Blood samples were also obtained from the horses every 14 days and macroclimatic data were obtained for the study period. Infestations of A. cajennense demonstrated distinct peaks of activity for each of the three parasitic stages over each 12-month period, showing a 1-year generation pattern. Larvae predominated from April to July and nymphs from June to October. Adults predominated from October to March with a greater number of adult males than females. Although other studies on seasonal dynamics in the states of Rio de Janeiro and Minas Gerais were performed with the free-living stages of A. cajennense on pastures, the present study in the state of São Paulo, performed with the parasitic stages of A. cajennense on horses, showed similar results to those observed in other states. Infestations by A. nitens demonstrated distinct peaks of activity of adult females (>4 mm), suggesting different tick generations during the year. Infestation with A. nitens was much higher in the first year than the second year which may have been related to horse nutritional status and stocking rate. Although several climatic variables showed statistical significant correlation (r) with tick counts, the determination coefficients (R(2)) were always lower than 0.40, suggesting that any single significant variable (i.e. mean temperature) would not explain the tick distribution pattern over the year. The highest peaks of A. nitens females (>4 mm) were significantly associated with decrease in horse packed cell volumes (R(2)=0.603). The ears and the perineum, tail and groin region accounted for around 70% of all A. nitens females counted on the horses.