Abstract: The present study described the morphological characters of Blepharipoda liberate and Lopoomastix japonica, and analyzed the genetic diversity of two species based on COⅠ gene. The diagnostic characters of B. liberate are as following:Carapace approximately 1.3 times longer than wide; strongly keeled medially; anterior margin with large rugosi spine on either side of ocular sinus; frontal region smooth with few low teeth posterior to rostrum. The diagnostic characters of L. japonica are as following:Carapace approximately 1.2 times longer than wide; gently keeled medially; anterior margin with proximally broad and distally narrowing rugose outer-ocular spines on either side of ocular sinus, concave medially. The gene diversity and nucleotide diversity of B. liberate based on COⅠ gene were 0.942±0.043 and 0.0059±0.0034, respectively. The gene diversity and the nucleotide diversity of L. japonica were 0.941±0.0048 and 0.0061±0.0035, respectively. The K-2P distance was 0.2705 between the B. liberate and L. japonica, and these two species were distributed into two distinct branches based on neighbor-joining tree analysis, which reinforces the validity of DNA barcoding on classification of Blepharipodidae based on COⅠ gene.