Fig 1.
Geographic location of Hubei Province, China.
Collection sites in this study are marked as triangles.
Table 1.
Nucleotide sequences of primers and conditions used for PCR amplification.
Fig 2.
Medicinal leeches (Hirudinaria sp.) collected in this study.
Fig 3.
Maximum likelihood phylogenetic tree of leech species based on the 18S rRNA gene.
The tree was constructed with the 18S rRNA sequences (926bp) by using the Kimura 2-parameter model with MEGA 7.0 (http://www.megasoftware.net); we rooted using Phyllidiella pustulosa sequence from Genbank and calculated bootstrap values with 1,000 replicates. The representative sequence of leeches in this study are in bold print and marked by circle. Scale bar indicates nucleotide substitutions per site.
Fig 4.
Maximum likelihood phylogenetic tree of leech species based on the COI gene.
The tree was constructed with the COI sequences (682bp) by using the Kimura 2-parameter model with MEGA 7.0 (http://www.megasoftware.net); we calculated bootstrap values with 1,000 replicates. The representative sequence of leeches in this study are in bold print and marked by circle. Scale bar indicates nucleotide substitutions per site.
Fig 5.
Maximum likelihood phylogenetic tree based on the rrs gene of Ehrlichia.
The tree was constructed with the rrs sequences (923bp) by using the general time-reversible model with MEGA 7.0 (http://www.megasoftware.net); we rooted using Anaplasma phagocytophilum sequence from Genbank and calculated bootstrap values with 1,000 replicates. The representative sequences obtained in this study are in bold print and marked by circles. Scale bar indicates nucleotide substitutions per site.
Fig 6.
Maximum likelihood phylogenetic tree based on the groEL gene of Ehrlichia.
The tree was constructed with the groEL sequences (422bp) by using the Tamura 3-parameter model with MEGA 7.0 (http://www.megasoftware.net); we rooted using Anaplasma sp. sequence from Genbank and calculated bootstrap values with 1,000 replicates. The representative sequence obtained in this study are in bold print and marked by circle. Scale bar indicates nucleotide substitutions per site.
Table 2.
Infection rate of Ehrlichia in leeches in different months from two farms.
Table 3.
Animal hosts of leeches determined by PCR using mtDNA cytochrome b gene from leeches.