The order Piroplasmida contains a diverse group of intracellular parasites, many of which can cause significant disease in humans, domestic animals, and wildlife. Two piroplasm species have been reported from raccoons ( Procyon lotor ), Babesia lotori ( Babesia sensu stricto clade) and a species related to Babesia microti (called B. microti -like sp.). The goal of this study was to investigate prevalence, distribution, and diversity of Babesia in raccoons. We tested raccoons from selected regions in the United States and Canada for the presence of Babesia sensu stricto and Babesia microti -like sp. piroplasms. Infections of Babesia microti -like sp. were found in nearly all locations sampled, often with high prevalence, while Babesia sensu stricto infections had higher prevalence in the Southeastern United States (20–45% prevalence). Co-infections with both Babesia sp. were common. Sequencing of the partial 18S rRNA and cytochrome oxidase subunit 1 ( cox1 ) genes led to the discovery of two new Babesia species, both found in several locations in the eastern and western United States. One novel Babesia sensu stricto sp. was most similar to Babesia gibsoni while the other Babesia species was present in the ‘western piroplasm’ group and was related to Babesia conradae . Phylogenetic analysis of the cox1 sequences indicated possible eastern and western genetic variants for the three Babesia sensu stricto species. Additional analyses are needed to characterize these novel species; however, this study indicates there are now at least four species of piroplasms infecting raccoons in the United States and Canada ( Babesia microti -like sp., Babesia lotori, a novel Babesia sensu stricto sp., a novel western Babesia sp.) and a possible fifth species ( Babesia sensu stricto) in raccoons in Japan.

Piroplasms are intraerythrocytic parasites that are often transmitted by ixodid ticks, but vertical transmission is an alternative route for some species. In the USA, raccoons (Procyon lotor) are hosts for two known species, a Babesia microti-like sp. and Babesia lotori (in Babesia sensu stricto group). To better understand the natural history of Babesia in raccoons, we tested young raccoons from Minnesota and Colorado for Babesia spp., examined them for ticks, and assessing for splenomegaly as a sign of clinical disease. Raccoons from both states were infected with B. microti-like sp. and Babesia sensu stricto spp. Infections of B. microti-like were common, even in 1-week-old raccoons, suggesting vertical transmission. Babesia sensu stricto infections were more common in older raccoons. Raccoons infected with Babesia sensu stricto had significantly higher spleen:body weight ratios compared with uninfected or B. microti-like sp.-infected raccoons. Ticks were only found on raccoons from Minnesota. The most common and abundant tick was Ixodes texanus but Ixodes scapularis and Dermacentor variabilis were also found on raccoons. We report piroplasm infections and infestations with several tick species in very young raccoons. Young raccoons infected with Babesia sensu stricto spp. had higher spleen:body weight ratios, suggesting a disease risk.