The transmission of parasites can be influenced by their co-occurrence with other parasites, in some cases increasing or reducing transmission. Trypanosoma cruzi, aetiologic agent of Chagas disease, often co-occurs with Trypanosoma rangeli, a parasite not pathogenic for mammal hosts. Both parasites can reduce the fitness of their insect vectors (the triatomine bugs; Hemiptera: Reduviidae), with T. rangeli being more pathogenic for some species. Here, we study the prevalence of T. cruzi and T. rangeli in the triatomine Rhodnius pallescens across a heterogeneously transformed landscape in Panamá. We found that single T. rangeli infections were more common in contiguously forested habitats, while single T. cruzi infections predominated in anthropogenically disturbed habitats. Trypanosoma cruzi–T. rangeli co-infections were more common in contiguous forests and in peridomiciliary areas. Furthermore, adult insects were more likely to be co-infected than nymphs. Our results suggest that human-mediated landscape transformation might have increased the predominance of single infections with T. cruzi within vectors. An important mechanism driving changes in trypanosome infection patterns in triatomines at a landscape scale includes alterations in host species composition that may vary with different degrees of deforestation. Trypanosome co-infection may also confer a survival advantage for R. pallescens to and/or throughout adulthood.