| Summary: | Recent years have witnessed an increase in the population of <i>Lipoptena</i><i>fortisetosa</i> in Central Europe. The genetic profile of this ectoparasite has not been studied in Poland to date. The aim of the present study was to confirm the presence of <i>L.</i><i>fortisetosa</i> in north-eastern Poland and to characterize the examined population with the use of molecular methods. Deer keds were collected between June and July 2019 in six natural, mixed forests. A fragment of the rRNA 16S gene was used as a marker to identify <i>L.</i><i>fortisetosa</i> by polymerase chain reaction (PCR). DNA samples were sequenced in the last step. Six new locations of <i>L. fortisetosa</i> were confirmed. No significant differences were observed in the sex ratios of <i>L. cervi</i> and <i>L. fortisetosa</i> (<i>L. cervi p</i>-value = 0.74; <i>L. fortisetosa p</i>-value = 0.65). Significant differences were noted between the total size of <i>L. cervi</i> and <i>L. fortisetosa</i> populations (<i>p</i>-value < 0.001). The similarity to GenBank sequences ranged from 95.56% to 100%. The obtained nucleotide sequences were very closely related to <i>L. fortisetosa</i> sequences from Lithuania, the Czech Republic, and Japan. Molecular analyses revealed considerable genetic diversity, which could indicate that various ectoparasite lineages have spread throughout Europe.
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