The complete mitochondrial genome of Echinostoma miyagawai: Comparisons with closely related species and phylogenetic implications.

Echinostoma miyagawai (Trematoda: Echinostomatidae) is a common parasite of poultry that also infects humans. Es. miyagawai belongs to the "37 collar-spined" or "revolutum" group, which is very difficult to identify and classify based only on morphological characters. Molecular techniques can resolve this problem. The present study, for the first time, determined, and presented the complete Es. miyagawai mitochondrial genome. A comparative analysis of closely related species, and a reconstruction of Echinostomatidae phylogeny among the trematodes, is also presented. The Es. miyagawai mitochondrial genome is 14,416 bp in size, and contains 12 protein-coding genes (cox1-3, nad1-6, nad4L, cytb, and atp6), 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs), and one non-coding region (NCR). All Es. miyagawai genes are transcribed in the same direction, and gene arrangement in Es. miyagawai is identical to six other Echinostomatidae and Echinochasmidae species. The complete Es. miyagawai mitochondrial genome A + T content is 65.3%, and full-length, pair-wise nucleotide sequence identity between the six species within the two families range from 64.2-84.6%. The Es. miyagawai sequences is most similar to Echinostoma caproni. Sequence difference are 15.0-33.5% at the nucleotide level, and 8.6-44.2% at the amino acid level, among the six species, for the 12 protein-coding genes. ATG and TAG are the most common initiation and termination codons, respectively. Twenty of the Es. miyagawai transfer RNA genes transcribe products of the conventional cloverleaf structure, while two of the transfer RNA genes, namely trnS1 and trnS2, have unpaired D-arms. Phylogenetic analyses using our mitochondrial data indicate that Es. miyagawai is closely related to other Echinostomatidae species, except for Echinostoma hortense, which forms a distinct paraphyletic branch, and Echinochasmus japonicus, which is outside the clade containing all other Echinostomatidae species. These phylogenetic results support the elevation of subfamily Echinostomatidae. Our dataset also provides a significant resource of molecular markers to study the taxonomy, population genetics, and systematics of the echinostomatids.