Nutria (Myocastor coypus) is a large rodent native to South America which was introduced worldwide originally with the intention of fur farming. Three colour forms (Standard, Moravian Silver, and Prestice Multicolour) fall into the Farm Animal Genetic Resources of the Czech Republic protected by the National Programme on Conservation and Utilization of Plant, Animal and Microbial Genetic Resources for Food and Agriculture coordinated by the National Focal Point. The objective of our investigation in the Czech Republic was to establish microsatellite panel for nutria and determine microsatellite variability in the local nutria population, levels of genetic diversity within and among subpopulations with respect to colour form and the levels of inbreeding within subpopulations. The genetic variability was studied in 64 animals by investigating 11 microsatellite markers (McoD214, McoD217, McoD59, McoD69, McoC124, McoC203, McoD60, McoB17, McoC118, McoA04, and McoD228) analysed by multicoloured capillary electrophoresis. The whole population showed a moderate level of genetic variability in terms of number of alleles (5 alleles per locus) or heterozygosity (50.4%) and the Prestice Multicolour form exhibited the highest level of diversity. Particular attention should be paid to organizing mate to minimise inbreeding, especially in the Standard colour form (F-IS = 0.238) which showed the highest level of inbreeding out of the three colour forms. High combined exclusion probability (CEP) values (99.5, 94.5, and 99.9%) implied that the panel of microsatellite markers established in this study was usable for individual identification or routine parentage testing in nutria population in the Czech Republic.