The prevalence of sparganum infection in wild frogs (Rana rugulosa and R. temporaria) and snakes (Enhydris chinensis) was investigated in Guangxi Zhuang Autonomous Region of Southern China from June 2013 to August 2013. The results showed that 30.37% (65/214) of the caught wild frogs and 50% (3/6) of examined snakes were found to be infected with plerocercoids (spargana) of the genus Spirometra. To investigate the genetic variation of the collected spargana, three mitochondrial DNA (mtDNA) regions, namely cytochrome c oxidase subunit 1 and 3 (cox1 and cox3), NADH dehydrogenase subunits 4 (nad4) were amplified, sequenced, and analyzed. The sequences of cox1, cox3 and pnad4 are 444, 387 and 585 bp in length, respectively. The base composition of cox1, cox3 and pnad4 were generally AT rich with a mean of 62.7%, 67.2% and 64.9%, respectively. Phylogenetic analyses using Bayesian inference (BI) and maximum parsimony (MP) indicated that all the Spirometra sparganum isolates in Guangxi represented S. erinaceieuropaei.