Abstract
Seven isolates of Cryptocaryon irritans from different host species and geographical locations in Mainland China were characterized by the first (ITS-1) and second (ITS-2) internal transcribed spacers (ITS) of nuclear ribosomal DNA (rDNA) using two isolates of Ichthyophthirius multifiliis for comparative purposes. The rDNA region including the ITS-1, 5.8S, ITS-2, and flanking 18S and 28S sequences were amplified by polymerase chain reaction and the amplicons were sequenced directly. The ITS-1, 5.8S, and ITS-2 sequences were 129, 160, and 190 bp in length, respectively, for all seven C. irritans isolates, whereas the corresponding sequences for the two I. multifiliis isolates were 142, 153, and 194 bp, respectively. While sequence variation among the seven C. irritans isolates ranged from 0 to 1.6% in both the ITS-1 and ITS-2, and the two I. multifiliis isolates differed by 1.4% in the ITS-1 and 1.0% in the ITS-2; C. irritans differed from I. multifiliis by 57.1–60.9% in the ITS-1 and 79.4–83.0% in the ITS-2, indicating that ITS sequences provide reliable genetic markers for the identification and differentiation of the two species. Phylogenetic analysis using the sequence pairwise-distance data using the neighbor-joining method inferred that the seven C. irritans isolates from Mainland China and two other isolates (T.A and Aus.C) from other countries clustered together to show monophyly, which could be readily distinguished from the other monophyletic group all from other regions. Therefore, ITS sequence data and phylogenetic analysis provided strong support that C. irritans isolates from Mainland China represent a single species. The definition of genetic markers in the ITS rDNA provide opportunities for studying the ecology and population genetic structures of the C. irritans from Mainland China and elsewhere and is also relevant to the diagnosis and control of fish diseases they cause.
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Acknowledgements
This project was supported and provided, in part, by grants from the National Natural Science Foundation of China (grant no. 30440052), the Natural Science Foundation of Guangdong Province (grant nos. 20031640 and 20023002), the Scientific and Technological Programmes of Zhuhai City (grant no. 200411022) to AXL, and from the China National Science Funds for Distinguished Young Scientists (grant no. 30225033) to XQZ. The experiments comply with the current laws of the countries in which the experiments were performed.
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Sun, H.Y., Zhu, X.Q., **e, M.Q. et al. Characterization of Cryptocaryon irritans isolates from marine fishes in Mainland China by ITS ribosomal DNA sequences. Parasitol Res 99, 160–166 (2006). https://doi.org/10.1007/s00436-006-0151-x
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DOI: https://doi.org/10.1007/s00436-006-0151-x