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Jawed polychaetes evolved and diversified extensively during the Ordovician. However, Ordovician polychaete jaws (scolecodonts) have remained poorly documented for many regions. This applies for the Prague Basin of peri-Gondwana, from where the previous study on Late Ordovician scolecodonts was published more than 70 years ago, with just two species preliminarily identified. The aim of the present paper was to fill this research gap and to study organic-walled microfossils from the boundary interval of the Králův Dvůr and Kosov formations (uppermost Katian and lowermost Hirnantian) at the Levín locality. As a result, a diverse assemblage of scolecodonts and chitinozoans was discovered. Chitinozoans are represented by at least 24 species from 15 genera, i.e., a relatively diverse assemblage whose species composition points toward the Ancyrochitina merga and Tanuchitina elongata biozones. The recovered jawed polychaete fauna contains at least 19 species from 14 genera. Taxa with labidognath and prionognath type maxillary apparatuses predominate in samples, whereas placognath and ctenognath taxa are relatively rare. A similar pattern is typical for the Laurentian samples but contrasts with the Baltic polychaete faunas. Polychaetaspids dominate in the Levín assemblage, followed by other families such as ramphoprionids, paulinitids, and atraktoprionids. The studied interval in the Levín section is represented by a succession of thin-bedded silty shales with diamictite beds, practically devoid of shelly fossils and with a variable degree of bioturbation. The deposits are interpreted as distal turbidites and debrites, reflecting sea-level changes driven by the growth and retreat of glacial ice and possibly also local tectonics. Reduced diversity and abundance of scolecodonts was recorded in the uppermost part of the Králův Dvůr Formation, which correlates with less bioturbation and finer silt fraction. The reported discovery shows wide geographical distribution and diversity of jawed polychaetes before and during the Hirnantian glaciation and mass extinction.