Background: The persistence of Triatoma infestans and the continuous transmission of Trypanosoma cruzi in the Inter-Andean Valleys and in the Gran Chaco of Bolivia are of great significance. Coincidentally, it is in these regions the reach of the vector control strategies is limited, and reports of T. infestans resistance to insecticides, including in wild populations, have been issued. This study aims to characterize the susceptibility to deltamethrin of wild and domestic populations of T. infestans from Bolivia, in order to better understand the extent of this relevant problem. Methods: Susceptibility to deltamethrin was assessed in nine, wild and domestic, populations of T. infestans from the Gran Chaco and the Inter-Andean Valleys of Bolivia. Serial dilutions of deltamethrin in acetone (0.2 μL) were topically applied in first instar nymphs (F1, five days old, fasting, weight 1.2 ± 0.2 mg). Dose response results were analyzed with PROBIT version 2, determining the lethal doses, slope and resistance ratios (RR). Qualitative tests were also performed. Results: Three wild T. infestans dark morph samples of Chaco from the Santa Cruz Department were susceptible to deltamethrin with RR 50 of <2, and 100% mortality to the diagnostic dose (DD); however, two domestic populations from the same region were less susceptible than the susceptibility reference lineage (RR 50 of 4.21 and 5.04 respectively and 93% DD). The domestic population of Villa Montes from the Chaco of the Tarija Department presented high levels of resistance (RR 50 of 129.12 and 0% DD). Moreover, the domestic populations from the Valleys of the Cochabamba Department presented resistance (RR 50 of 8.49 and 62% DD), the wild populations were less susceptible than SRL and T. infestans dark morph populations (RR 50 < 5). Conclusion: The elimination of T. infestans with pyrethroid insecticides in Brazil, Uruguay, Chile, and its drastic reduction in large parts of Paraguay and Argentina, clearly indicates that pyrethroid resistance was very uncommon in non-Andean regions. The pyrethroid susceptibility of non-Andean T. infestans dark morph population, and the resistance towards it, of Andean T. infestans wild and domestic populations, indicates that the Andean populations from Bolivia are less susceptible.