During the spring of 2014, a wide survey was conducted in one of the most important mango (Mangifera indica) cultivating areas located in Minas Gerais State (Brazil) to ascertain the causal agent of severe anthracnose infections and to evaluate disease susceptibility within a world collection of mango germplasm. Overall, 86 cultivars were monitored and 152 fungal isolates recovered from infected samples were identified by morphological characterization, DNA sequencing and phylogenetic analyses. All isolates were identified as Colletotrichum asianum. Under natural disease pressure, it has been possible to ascertain a variable tolerance degree within the germplasm collection. By applying a categorized classification, cultivars were classified as follows: 10 highly sensitive (11.6%), 13 sensitive (15.1%), 18 moderately sensitive (20.9%), 23 moderately tolerant (26.7%), 11 tolerant (12.8%), and 11 highly tolerant (10.4%). The most susceptible cultivars to anthracnose were Ubà, Quinzenga, Amarelinha da Sementeira followed by Aroeira and Correjo, whereas Mallika followed by Ourinho and Lita resulted in the least susceptible cultivars. To the authors’ knowledge, this is the first large-scale evaluation of mango susceptibility to C. asianum infections within a wide number of cultivars. Anthracnose is a serious threat to mango production and assessment of cultivar response to disease could be useful in breeding programs.