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In this paper we describe Macrobiotus engbergi sp. nov. and Tenuibiotus zandrae sp. nov. from Greenland. Our study has involved both classical taxonomic methods, which include morphological and morphometric analyses conducted with the use of light and scanning electron microscopy, and genetic analysis based on four molecular markers (three nuclear: 18S rRNA, 28S rRNA, ITS-2, and one mitochondrial: COI). Moreover, we re-examined the type series of Tenuibiotus voronkovi (Tumanov, 2007) as well as the original sample where the species was found and we provide new morphological data from light and scanning electron microscopy which enabled us to amend its description. Finally, we also analysed slides with animals and egg of two populations from Nordaustlandet and Edgeøya (archipelago of Svalbard, Norway) designated as T. voronkovi within its recent redescription. The results and comparisons presented in our study question the validity of this designation.
In this study, I describe two new species of Macrobiotus based on morphological data collected through light and scanning electron microscopy. Both species are accompanied by DNA sequences from four commonly used molecular markers (18S rDNA, 28S rDNA, ITS-2, and COI). Macrobiotus ovovittatus sp. nov. was discovered in Greenland and can be distinguished from similar taxa of Macrobiotus by its continuous, solid, and clearly wrinkled egg surface, adorned with sparse, very small and irregularly spaced pores. Additionally, the terminal discs of egg processes are covered in multiple light-refracting dots, resembling crocheted napkins. Macrobiotus mileri sp. nov. was found in Israel and is characterized by unique pore arrangements in its body cuticle, expressed in two distinct animal forms: (i) forma porata with large pores arranged in five distinct patches and (ii) forma aporata with single, almost undetectable pores. It also features weakly defined convex terminal discs with smooth edges. Furthermore, the phylogenetic analyses conducted in this study offer the most updated phylogeny of superclade I within the family Macrobiotidae. This facilitates additional discussion concerning the interrelationships among species within the genus Macrobiotus and the circumscription of species groups within it.