<i>Spirometra</i> tapeworms (Cestoda: Diphyllobothriidae) collected from carnivorous mammals in Tanzania were identified by the DNA sequence analysis of the mitochondrial cytochrome c oxidase subunit 1 (<i>cox</i>1) and internal transcribed spacer 1 (ITS1), and by morphological characteristics. A total of 15 adult worms were collected from stool samples and carcasses of <i>Panthera leo, Panthera pardus</i>, and <i>Crocuta crocuta</i> in the Serengeti and Selous ecosystems of Tanzania. Three <i>Spirometra</i> species: <i>S. theileri, S. ranarum</i> and <i>S. erinaceieuropaei</i> were identified based on morphological features. Partial <i>cox</i>1 sequences (400 bp) of 10 specimens were revealed. Eight specimens showed 99.5% similarity with <i>Spirometra theileri</i> (MK955901), 1 specimen showed 99.5% similarity with the Korean <i>S. erinaceieuropaei</i> and 1 specimen had 99.5% similarity with Myanmar <i>S. ranarum</i>. Sequence homology estimates for the ITS1 region of <i>S. theileri</i> were 89.8% with <i>S. erinaceieuropaei</i>, 82.5% with <i>S. decipiens</i>, and 78.3% with <i>S. ranarum</i>; and 94.4% homology was observed between <i>S. decipiens</i> and <i>S. ranarum</i>. Phylogenetic analyses were performed with 4 species of <i>Spirometra</i> and 2 species of <i>Dibothriocephalus</i> (=<i>Diphyllobothrium</i>). By both ML and BI methods, <i>cox</i>1 and ITS1 gave well supported, congruent trees topology of <i>S. erinaceieuropaei</i> and <i>S. theileri</i> with <i>S. decipiens</i> and <i>S. ranarum</i> forming a clade. The <i>Dibothriocephalus</i> species were sisters of each other and collectively forming successive outgroups. Our findings confirmed that 3 <i>Spirometra</i> species (<i>S. theileri, S. ranarum</i>, and <i>S. erinaceieuropaei</i>) are distributed in the Serengeti and Selous ecosystems of Tanzania.