The occurrence of hypertetraploid and other unusual polyploid loaches Misgurnus anguillicaudatus among market specimens in Japan

Exotic animals can cause genetic contamination of indigenous species if they escape and reproduce in wild populations. The loach Misgurnus anguillicaudatus and species related to it have been imported into Japan for commercial use. We collected live loach specimens from the central wholesale market in Tokyo. For the 451 specimens collected, ploidy status was examined by DNA content flow cytometry, and polyploid loaches with triploid, tetraploid, and other higher ploidy ranges were detected. Hypertriploid and hypertetraploid individuals were easily detected by flow cytometry using standard eudiploid, eutriploid, and eutetraploid controls, and the reproductive capacities of these hyperpolyploid males were then examined. Sperm of hypertriploid males did not exhibit active progressive motility, and major populations of spermatozoa or spermatozoon-like cells were detected in the triploid and hexaploid ranges. Motile haploid spermatozoa were very rare in sperm from hypertriploid males. Therefore, hypertriploid males were sterile, whereas hypertetraploid males produced fertile hyperdiploid spermatozoa with active progressive motility after contact with ambient water. Viable progeny occurred in crosses between normal wild-type diploid females and hypertetraploid males, but androgenotes induced by the fertilization of UV-irradiated eggs with the sperm of hypertetraploid males were inviable hyperdiploids. Cytogenetic analyses of those androgenotes indicated that hypertetraploid males should produce hyperdiploid spermatozoa with 2n = 54 (i.e., four supernumerary microchromosomes).