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Among the tardigrades, distinguishing between species from the 'hufelandi group' and between species from genus Milnesium is based primarily on morphological data. These are difficult and time‐consuming to obtain and are very often not sufficient. In this study, we promote the ITS2 marker as the most suitable tool for distinguishing closely related species with low morphological diversity. We present new molecular records of Macrobiotus polonicus and Milnesium tardigradum to validate their current phylogenetic position within Tardigrada and to resolve, in further analyses, the phylogenetic relationships within the 'hufelandi group' and among species from the genus Milnesium. Moreover, we provide a reliable method to obtain the complete 18S rDNA and ITS2 sequences. The present study includes also the analyses of ITS2 secondary structures. [ABSTRACT FROM AUTHOR]

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