Molecular identification of the species of Biomphalaria snails in Shenzhen reservoir based on 16S rDNA sequences

Abstract

Objective To identify the species of Biomphalaria snails collected in Shenzhen reservoir, based on the mitochondrial 16S rDNA sequences. Methods The 16S rDNA fragments were amplified by PCR from the genome DNA of Biomphalaria snails, and inserted in plasmid pMD-18T for sequencing. The sequence of 16S rDNA fragment and its phylogenetic relationships with those of other species of Biomphalaria snails were analyzed with BLAST and MEGA4 software. Results The amplified 16S rDNA fragment of the Biomphalaria snails was about 466 bp in length. As aligned with the corresponding sequences of the related Biomphalaria species, the identity of nucleotides was 99% with 1 isolate of Biomphalaria straminea (B. straminea), 98% with 3 isolates of B. kuhniana, 95% with 1 isolate of B. intermedia, and 94% with 1 isolate of B. edisoni. Based on the 16S rDNA sequence, the results of phylogenetic analysis with neighbor-joining (NJ) and unweighted pair-group method with arithmetic means (UPGMA) indicated that the snails had close genetic relationships with the B. straminea isolate(Genbank accession NO. AY030213.1) Conclusion The Biomphalaria snails collected in Shenzhen reservoir could be classified as B. straminea based on the characteristics of 16S rDNA sequence. Key words: Biomphalaria snails; DNA, Mitochondrial; Molecular phylogenetic analysis