New insights into the evolution of the Trypanosoma cruzi clade provided by a new trypanosome species tightly linked to Neotropical Pteronotus bats and related to an Australian lineage of trypanosomes

Abstract\ud \ud Background\ud Bat trypanosomes are implicated in the evolution of the T. cruzi clade, which harbours most African, European and American trypanosomes from bats and other trypanosomes from African, Australian and American terrestrial mammals, including T. cruzi and T. rangeli, the agents of the American human trypanosomiasis. The diversity of bat trypanosomes globally is still poorly understood, and the common ancestor, geographical origin, and evolution of species within the T. cruzi clade remain largely unresolved.\ud \ud \ud Methods\ud Trypanosome sequences were obtained from cultured parasites and from museum archived liver/blood samples of bats captured from Guatemala (Central America) to the Brazilian Atlantic Coast. Phylogenies were inferred using Small Subunit (SSU) rRNA, glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH), and Spliced Leader (SL) RNA genes.\ud \ud \ud Results\ud Here, we described Trypanosoma wauwau n. sp. from Pteronotus bats (Mormoopidae) placed in the T. cruzi clade, then supporting the bat-seeding hypothesis whereby the common ancestor of this clade likely was a bat trypanosome. T. wauwau was sister to the clade T. spp-Neobats from phyllostomid bats forming an assemblage of trypanosome species exclusively of Noctilionoidea Neotropical bats, which was sister to an Australian clade of trypanosomes from indigenous marsupials and rodents, which possibly evolved from a bat trypanosome. T. wauwau was found in 26.5 % of the Pteronotus bats examined, and phylogeographical analysis evidenced the wide geographical range of this species. To date, this species was not detected in other bats, including those that were sympatric or shared shelters with Pteronotus. T. wauwau did not develop within mammalian cells, and was not infective to Balb/c mice or to triatomine vectors of T. cruzi and T. rangeli.\ud \ud \ud Conclusions\ud \ud Trypanosoma wauwau n. sp. was linked to Pteronotus bats. The positioning of the clade T. wauwau/T.spp-Neobats as the most basal Neotropical bat trypanosomes and closely related to an Australian lineage of trypanosomes provides additional evidence that the T. cruzi clade trypanosomes likely evolved from bats, and were dispersed in bats within and between continents from ancient to unexpectedly recent times.This work was supported by grants from the Brazilian agencies CNPq (PROSUL,\ud PRAFRICA and PROTAX), CAPES (PNIPB and PNPD) and FAPESP. The analysis of\ud bats from Central America, Suriname and Guyana was supported by grant\ud ‘Investissements d’Avenir’ from the Agence Nationale de la Recherche, Canada\ud (ANR-10-LABX-25-01). Archived samples from Brazilian Pteronotus were donated\ud to ACP by VC Tavares, A Césari, PA Rocha, FM Martins, MOG Lopes, CS Bernabé,\ud TG Oliveira, E Gonçalves and M Marcos. We are grateful to many student from\ud USP and researches of other universities for the inestimable help in the\ud fieldworks. We also thanks JA Rosa for the generous contribution with\ud triatomines from the insectary of UNESP-Araraquara, and CE Jared and MM\ud Antoniazzi for the access to electron microscopic facilities of the Institute\ud Butantan, Brazil. Luciana Lima is postdoctoral fellow sponsored by FAPESP, and\ud Oneida Espinosa-Álvarez is recipient of a PhD fellowship from CNPq (PROTAX)

Additional file 1: Table S1. of New insights into the evolution of the Trypanosoma cruzi clade provided by a new trypanosome species tightly linked to Neotropical Pteronotus bats and related to an Australian lineage of trypanosomes

Prevalence of Trypanosoma wauwau and geographical origin of Pteronotus spp. examined in this study. (DOC 114 kb