A new Myotis is described from the Islas Tres Marias, Nayarit, Mexico. the new species is distinct from related taxa n the adjacent Mexican mainland (M. californicus, M. leibii, and M. carteri), although most closely related to M. carteri as shown by univariate and canonical variates analyses. An analysis of six groups of M. nigricans from Middle and South America supports the elevation of M. nigricans carteri to specific status, confirms the distinctness of M. nigricus extremus, but fails to substantiate subspecific status for bats from Columbia and Ecuador, recent recognized as M. n. punensis.