| Summary: | Triazole-resistance has been reported increasingly in <i>Aspergillus fumigatus</i>. An international expert team proposed to avoid triazole monotherapy for the initial treatment of invasive aspergillosis in regions with >10% environmental-resistance, but this prevalence is largely unknown for most American and African countries. Here, we screened 584 environmental samples (soil) from urban and rural locations in Mexico, Paraguay, and Peru in Latin America and Benin and Nigeria in Africa for triazole-resistant <i>A. fumigatus</i>. Samples were screened using triazole-containing agars and confirmed as triazole-resistant by the European Committee on Antimicrobial Susceptibility Testing (EUCAST) broth dilution reference method. Isolates were further characterized by <i>cyp51A </i>sequencing and short-tandem repeat typing. Fungicide presence in samples was likewise determined. Among <i>A. fumigatus</i> positive samples, triazole-resistance was detected in 6.9% (7/102) of samples in Mexico, 8.3% (3/36) in Paraguay, 9.8% (6/61) in Peru, 2.2% (1/46) in Nigeria, and none in Benin. <i>C</i><i>yp51A</i> gene mutations were present in most of the triazole-resistant isolates (88%; 15/17). The environmentally-associated mutations TR<sub>34</sub>/L98H and TR<sub>46</sub>/Y121F/T289A were prevalent in Mexico and Peru, and isolates harboring these mutations were closely related. For the first time, triazole-resistant <i>A. fumigatus</i> was found in environmental samples in Mexico, Paraguay, Peru, and Nigeria with a prevalence of 7–10% in the Latin American countries. Our findings emphasize the need to establish triazole-resistance surveillance programs in these countries.
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