The genus Spirometra belongs to the order Pseudophyllidea and family Diphyllobothridea. The aim of the present study was to determine the complete life cycle of the tapeworm Spirometra and characterize the parasite using traditional parasitological methods and molecular biology techniques, and determine seroprevalence of spargana antibodies, knowledge, attitudes and practices on sparganosis infection among humans in Babati and Monduli districts, Tanzania. The investigation was conducted through targeted experimental life cycle, molecular biology, serology and questionnaire survey. Spirometra eggs were collected from faeces of lions and dogs from Tarangire National Park and Minjingu village. Eggs were cultured by modified Harada-Mori method. Hatched coracidia were experimentally fed to Cyclops to develop to procercoid. Infected Cyclops were orally fed to mice, rat, guinea pigs, New Zealand rabbit, pig and goat to develop to plerocercoids. The adult worm recovered from naturally infected dog from Minjingu village was used for morphological description after staining with Carmine. Polymerase Chain Reaction (PCR), DNA sequencing and phylogeny were used for identification and genetic characterization of the worm. A total of 216 serum samples obtained from normal inhabitants of Babati and Monduli districts were tested by enzyme-linked immunosorbent assay (ELISA) for anti Spirometra plerocercoid IgG. Questionnaire was used to collect information on knowledge, attitude and practices for sparganosis among inhabitants from the two selected districts. Coracidia hatched out of Spirometra eggs developed to procercoid in Cyclops. Plerocercoid was not obtained in both naturally and experimentally infected animals. Identification of Tanzanian Spirometra spp. using morphological studies and molecular techniques confirmed to be Spirometra erinaceieuropaei. Serum samples revealed 62.5% (n =135) positive anti Spirometra plerocercoid IgG. All (100%) participants (n = 345) had no knowledge about sparganosis and mode of transmission of the disease. This study revealed inadequate knowledge, attitudes and practices on iii sparganosis among inhabitants in the two districts. The health significance of Spirometra infection needs further investigation to establish relevance in health programmes in Tanzania.Mzumbe University