Characterization of tongue worms, Linguatula spp. (Pentastomida) in Romania, with the first record of an unknown adult Linguatula from roe deer (Capreolus capreolus Linnaeus)

Specimens of the pentastomid parasite, Linguatula serrata, have been reported from several animals in Romania, including some domestic dogs translocated to other parts of Europe. In this study, gray wolves (Canis lupus, n = 80), golden jackals (C. aureus, n = 115), red foxes (Vulpes vulpes, n = 236), and roe deer (Capreolus capreolus, n = 1) were examined for pentastomes. Overall, 17.5% of wolves were found to be infected with specimens of Linguatula, with a range of infections of one to five individuals per animal. Golden jackals and foxes had much lower infection levels, with 1.73% of golden jackals and 1.69% of foxes infected; both host species were found to be infected with one or two individual pentastomes per animal. The single deer specimen was infected with three individual pentastomes. The pentastomes collected from the wolves and golden jackals were determined to be immature and mature adult specimens of L. serrata based on morphological examination and molecular analysis using the 18S rRNA gene. No pentastomes collected from the red foxes were available for identification. The pentastomes collected from the roe deer were expected to be L. arctica but determined to be mature adult male specimens of an unknown Linguatula, herein, referred to as Linguatula sp. based on its morphology; the results of molecular sequencing for the Linguatula specimen collected from the deer were inconclusive, preventing a final species identification. This study presents the first report of L. serrata in any hosts from Romania through both morphological and molecular characterization, and also presents the first report of a Linguatula sp. in Ca. capreolus, utilizing morphological characterization. Issues of morphological variability are discussed, including the presence of spines in the hook pit of specimens of Linguatula. This study highlights the need to examine all specimens of Linguatula to confirm the stage of development. Despite the inconclusive molecular result for some specimens, the authors still urge future researchers to incorporate a combined molecular and morphological approach in identifying specimens of Linguatula.