Project Summary/ Abstract African tick-bite fever caused by Rickettsia africae is probably the most common human spotted fever group rickettsiosis. It occurs throughout Africa, wherever its major vector, Amblyomma variegatum, is found. Other Amblyomma species can also transmit R. africae and the organism has spread in Amblyomma loculosum on migratory birds through the Indian and Pacific Ocean islands. Rickettsia africae also occurs in Central America in A. ovale, and in the Caribbean islands in A. variegatum which were imported on cattle from Africa in the 1800s. Infected ticks were spread widely around the Caribbean islands on cattle egrets, which are migratory birds that move readily between the islands and as far as the Florida Keys. The presence of R. africae in the nearby Caribbean islands and Central America, and Africa, the expanding tourism and trade between these areas and the USA, the travelers entering the USA from Africa, the Caribbean, and Central America that are infected with R. africae, and the presence of migratory birds raises the risks of A. variegatum and/or R. africae being introduced into the USA and becoming established. If Amblyomma species endemic in the USA could then become infected with and maintain and transmit R. africae, African tick-bite fever could become established in the USA causing widespread morbidity as it does in Africa. Additionally, this would further complicate the already difficult diagnosis of American spotted fever group Rickettsia infections, in particular Rocky Mountain spotted fever. The effects of R. africae on American wildlife are unknown. To establish the reservoir and vector capacity of the most relevant US Amblyomma species, A. americanum and A. maculatum, we intend to conduct transmission experiments with guinea pigs and calves as hosts. Amblyomma variegatum larvae/ nymphs and adults infected with R. africae will be fed on guinea pigs or calves, respectively, at the same time as uninfected immature and adult US Amblyomma species to determine if infections can be transferred horizontally. If infections can be demonstrated in the US Amblyomma species, further experiments will be performed to establish if R. africae can be transmitted vertically between feeding stages (transtadially), transovarially through the eggs, and from each feeding stage to the host. Data generated from these experiments will enable American health workers to more precisely determine the risk of African tick-bite fever becoming established in the USA and for appropriate prevention and response strategies to be developed.