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We present a review on the natural infection by trypanosomatids of nonhuman vertebrates in Chile, aiming to synthesize and update the knowledge on the diversity of trypanosomatids infecting native and alien vertebrate species. To this end, we conducted a systematic review of literature records published from 1900 to April 2020 on four databases, focusing on the 21 genera of trypanosomatids and Chile. The methods and findings of our review have been based on the preferred reporting items for systematic reviews and meta-a