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In Europe wildlife animals such as the red fox (Vulpes vulpes) are considered the main reservoir for Angiostrongylus vasorum as well as a potential threat for domestic dog infection. Though this parasite is endemic in fox populations, data on A. vasorum infection in wolves (Canis lupus italicus) are still scant, having only recently been described in Northwestern Spain, in Italy, in Croatia and in Slovakia. Based on the rising number of cases of canine lungworm infection in Central Italy (Abruzzo region), the aim of the present study was to investigate the infection by A. vasorum in fox and wolf populations sharing the same geographical area of dogs. From October 2008 to November 2019, A. vasorum specimens were collected, through routine post-mortem examination, from 56 carcasses (44 foxes and 12 wolves). Adult parasites were searched for in the right side of the heart and in pulmonary artery of all carcasses. First stage of larvae (L1) was searched in faeces using the Baermann technique and in lungs by tissue impressions. Overall, 230 adult specimens were collected and identified on a morphological basis. To confirm the morphological identification, 4 adult specimens (n = 3 from fox, n = 1 from wolf) were molecularly identified as A. vasorum by amplification of partial fragment of nuclear 18S rRNA (~1700 bp) genes. The anatomo-pathological and parasitological examinations indicated the presence of A. vasorum in 33 foxes (75%) and in 8 wolves (66.7%). The level of prevalence of infested wolves was higher than the previous one reported in other European countries. Interestingly, the prevalence of infection in foxes herein recorded was higher than that described in dogs (8.9%) living in the same geographical area. This result may confirm the hypothesis that the spread of canine angiostrongylosis is linked to fox populations infection., Graphical abstract Image 1, Highlights • Angiostrongylus vasorum was found in dead foxes and wolves in Chieti province, Italy. • The prevalence in foxes was higher than that described in dogs. • The prevalence in wolves was higher than the previous one reported in European countries. • The spread of canine angiostrongylosis is linked to fox populations infection. • The province of Chieti appears to be an enzootic focus for canine angiostrongylosis.

Graphical abstract, Highlights • First report of Cercopithifilaria rugosicauda in a roe deer and ticks from Italy. • This study provides new morphological data on this little known nematode. • The genetic data provide for the first time information on mitochondrial genes of C. rugosicauda., Cercopithifilaria rugosicauda (Spirurida, Onchocercidae) is a subcutaneous filarial nematode of the European roe deer (Capreolus capreolus) transmitted by Ixodes ricinus (Acari, Ixodidae). At the necropsy of a roe deer from the Parco Regionale di Gallipoli Cognato (Basilicata region, southern Italy), two female nematodes of C. rugosicauda were found. Following the necropsy, seven skin snips were sampled from different body regions and 96 I. ricinus ticks were collected. In addition, 240 ticks were collected by dragging in the enclosure where the roe deer lived. Samples were examined for the presence of C. rugosicauda larvae and assayed by PCR targeting cytochrome c oxidase subunit 1 (cox1, ∼300 bp) and 12S rDNA (∼330 bp) gene fragments. Female nematodes, microfilariae from skin samples and eight third stage larvae (L3) from ticks were morphologically and molecularly identified as C. rugosicauda. Phylogenetic analyses clustered this species with other sequences of Cercopithifilaria spp. This study represents the first report of C. rugosicauda in a roe deer and ticks from Italy and provides new morphological and molecular data on this little known nematode.