Tetranematichthys barthemi is described from the mouth of the rio Trombetas and related tributaries (rios Saracá and Araticum) and rio Urubu, tributary of the rio Amazon. Tetranematichthys barthemi shares with its congeners the anterior alignment of the mandibular barbels and well-developed portion of the protractor hyoid muscle connected to the mandibular barbel. It is distinguished from these species by a serrated anterior margin on the distal portion of the mandibular barbel, different from the more derived state which is composed of conspicuous digitiform elaboration in the distal portion of the barbel. Tetranematichthys barthemi can also be distinguished from its congeners by combination of characters related to morphometry and coloration pattern. Comments on sexual dimorphism, changes to the extremities of the mandibular barbels and possible interspecific relationships are presented herein.