IntroductionTuberculosis is an infectious disease caused by a group of acid-fast bacilli known asMycobacterium tuberculosiscomplex (MTC), which has a major impact on humans. Transmission of MTC across the human-animal interface has been demonstrated by several studies. However, the reverse zoonotic transmission from humans to animals (zooanthroponosis) has often been neglected.MethodsIn this study, we used Nanopore MinION and Illumina MiSeq approaches to sequence the whole genome ofM. tuberculosisstrains isolated from two deceased Asian elephants (Elephas maximus) and one human in Chitwan, Nepal. The evolutionary relationships and drug resistance capacity of these strains were assessed using the whole genome data generated by the stand-alone tool Tb-Profiler. Phylogenomic trees were also constructed using a non-synonymous SNP alignment of 2,596 bp, including 94 whole genome sequences representative of the previously describedM. tuberculosislineages from elephants worldwide (lineages 1 and 4) and from humans in Nepal (lineages 1, 2 and 3).Results and DiscussionThe new genomes achieved an average coverage of 99.6%, with an average depth of 55.67x. TheseM. tuberculosisstrains belong to lineage 1 (elephant DG), lineage 2 (elephant PK) and lineage 4 (human), and none of them were found to have drug-resistant variants. The elephant-derived isolates were evolutionarily closely related to human-derived isolates previously described in Nepal, both in lineages 1 and 2, providing additional support for zooanthroponosis or bidirectional transmission between humans and elephants. The human-derived isolate clustered together with other published human isolates from Argentina, Russia and the United Kingdom in the lineage 4 clade. This complex multi-pathogen, multi-host system is challenging and highlights the need for a One Health approach to tuberculosis prevention and control at human-animal interface, particularly in regions where human tuberculosis is highly endemic.