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Dioctophyme renale , the giant kidney worm, is a nematode related to Trichuris sp and is distributed worldwide. These parasites locate in the kidney of their definitive hosts (mainly belonging to the order Carnivora) and have an indirect life cycle with an annelid as the main intermediate host. Humans are rarely affected, but in those that are, 1 or both kidneys are destroyed. In South America, D. renale is widespread in riparian regions where changing climatic conditions, environmental degradation, and compromised sanitation are increasing the risk of distribution of this parasite, including humans. Here, we provide the descriptions of the genetic diversity of the parasite in the region by analysing 73 adult D. renale samples collected from domestic and wild carnivores. The most common hosts were ( Canis lupus familiaris) and maned wolf ( Chrysocyon brachyurus Fam. Canidae) among domestic and wild carnivores, respectively. This work shows the descriptions of the genetic diversity of this parasite complementing molecular methods and classical and probabilistic phylogeography. Our results strongly suggest that this parasite has been present on the continent long enough to develop local genetic variants. Also, the phylogenies show transmission between localities and bidirectional transmission between domestic and wild species. We now have new tools to understand the ecological dynamics of this parasite such as molecular markers to study its genetic diversity as well as for identification and reporting in cryptic cases.

Echinococcosis, caused by cestodes of the genus Echinococcus , poses significant public health and veterinary concerns globally. In Armenia, cystic echinococcosis (CE) is well-documented in livestock and humans, while alveolar echinococcosis (AE) has long been considered non-endemic. However, a recent retrospective study identified human AE cases, suggesting an underestimation of the parasite’s presence. To address knowledge gaps, a pilot survey was conducted to identify Echinococcus species and other taeniids in free-roaming dogs and wild carnivores in Armenia. Fecal samples ( n = 112) were opportunistically collected from eight wild carnivore species and stray dogs across six Armenian provinces between 2017 and 2018. Samples were analysed for taeniid eggs using flotation and molecular techniques. Echinococcus multilocularis was identified in a free-roaming dog, marking the first confirmed detection of the parasite in a definitive host in Armenia. Additionally, E. canadensis G6/7 and E. ortleppi were detected in a wolf, while E. canadensis G6/7, Taenia hydatigena , and T. laticollis were found in a lynx. These findings indicate the involvement of both domestic and wild carnivores in the transmission cycles of Echinococcus species and suggest the presents of a potential sylvatic cycle involving E. canadensis G6/7 and E. ortleppi . The presence of E. multilocularis in a dog underscores the risk of human infection, necessitating further epidemiological studies. This study provides initial insights into the epidemiology of Echinococcus in Armenia and highlights the need for continued surveillance to assess public health risks.

Wild carnivores may be important sources of emerging zoonotic pathogens worldwide, however, specific data concerning their presence in large carnivores are limited. The genus Trypanosoma (protozoan parasites) comprises numerous species adapted to all classes of vertebrates. This genus includes highly pathogenic species, such as T. brucei ssp., T. congolense or T. vivax . The aim of this study was to expand on the ecological or zoonotic significance of detecting Trypanosoma in large carnivores. Samples from free-living carnivores ( n  = 26) were collected. The diversity and occurrence of Trypanosoma species among wildlife in Europe were investigated through the analysis of 18S rRNA gene sequences. The phylogenetic analysis showed three new haplotypes of the 18S rRNA gene of Trypanosoma sp. isolated from two grey wolves ( Canis lupus ) and two brown bears ( Ursus arctos ). To the best of our knowledge, this this study represents the first detection of trypanosomes in large carnivores in Europe. The detection of Trypanosoma sp. was achieved through nested- PCR amplification. Furthermore, the identification of new haplotypes of these protozoan highlights variability within Trypanosoma sp., which could lead to the emergence of new, potentially more virulent variants of these parasites in wild animal populations, posing a potential threat to their health. Moreover, these findings underscore the need for further research to comprehensively understand the diversity, distribution, and potential impact of Trypanosoma species on the populations of large carnivores.

Piroplasmids are tick-borne protozoan parasites that infect blood cells (erythrocytes, lymphocytes or other leukocytes) or endothelial cells of numerous wild and domestic vertebrates worldwide. They cause severe disease in livestock, dogs, cats, wild mammals and, occasionally, in humans. Piroplasmid infections are prevalent in wild carnivores worldwide although there is limited information about their clinical and epidemiological importance. There are currently nine recognized species of Babesia , two of Theileria , two of Cytauxzoon and one of Rangelia infecting captive and wild carnivores, including members of Canidae, Felidae, Mustelidae, Procyonidae, Ursidae, Viverridae, Hyaenidae and Herpestidae in the Americas, Eurasia and Africa. However, the number of piroplasmid species is likely higher than currently accepted due to the reported existence of DNA sequences that may correspond to new species and the lack of studies on many host species and biogeographical areas. Indeed, many species have been recognized in the last few years with the advancement of molecular analyses. Disease and mortality have been documented in some wild carnivores, whereas other species appear to act as natural, subclinical reservoirs. Various factors (e.g. unnatural hosts, stress due to captivity, habitat degradation, climate fluctuation or immunosuppression) have been associated with disease susceptibility to piroplasmid infections in some species in captivity. We aimed to review the current knowledge on the epidemiology of piroplasmid infections in wild carnivores and associated tick vectors. Emphasis is given to the role of wild carnivores as reservoirs of clinical piroplasmosis for domestic dogs and cats, and to the importance of piroplasmids as disease agents for endangered carnivores.

Most parasites infect multiple hosts, but what factors determine the range of hosts a given parasite can infect? Understanding the broad scale determinants of parasite distributions across host lineages is important for predicting pathogen emergence in new hosts and for estimating pathogen diversity in understudied host species. In this study, we used a new data set on 793 parasite species reported from free‐ranging populations of 64 carnivore species to examine the factors that influence parasite sharing between host species. Our results showed that parasites are more commonly shared between phylogenetically related host species pairs. Additionally, host species with higher similarity in biological traits and greater geographic range overlap were also more likely to share parasite species. Of three measures of phylogenetic relatedness considered here, the number divergence events that separated host species pairs most strongly influenced the likelihood of parasite sharing. We also showed that viruses and helminths tend to infect carnivore hosts within more restricted phylogenetic ranges than expected by chance. Overall, our results underscore the importance of host evolutionary history in determining parasite host range, even when simultaneously considering other factors such as host ecology and geographic distribution.

Background Hepatozoon spp. are tick-borne parasites causing subclinical to clinical disease in wild and domestic animals. Aim of this study was to determine Hepatozoon prevalence and species distribution among wild mammals and ticks in Europe. Methods Samples of wild mammals and ticks, originating from Austria, Bosnia and Herzegovina, Croatia, Belgium and the Netherlands, were tested with PCR to amplify a ~ 670-bp fragment of the small subunit ribosomal RNA gene. Results Of the 2801 mammal samples that were used for this study, 370 (13.2%) tested positive. Hepatozoon canis was detected in samples of 178 animals (3 Artiodactyla, 173 Carnivora, 1 Eulipotyphia, 1 Lagomorpha), H. martis in 125 (3 Artiodactyla, 122 Carnivora), H. sciuri in 13 (all Rodentia), Hepatozoon sp. in 47 (among which Hepatozoon sp. Vole isolate, all Rodentia) and H. ayorgbor in 4 (all Rodentia). Regarding origin, 2.9% (6/208) tested positive from Austria, 2.8% (1/36) from Bosnia and Herzegovina, 14.6% (173/1186) from Croatia and 13.9% (190/1371) from Belgium/the Netherlands. Of the 754 ticks collected, 0.0% (0/35) Hyalomma sp., 16.0% (4/25) Dermacentor spp., 0.0% (0/23) Haemaphysalis spp., 5.3% (24/50) Ixodes and 1.4% (3/221) Rhipicephalus spp. tested positive for Hepatozoon (4.2%; 32/754), most often H. canis ( n  = 22). Conclusions Hepatozoon canis is most present in mammals (especially in Carnivora such as gray wolves and golden jackals) and ticks, followed by H. martis , which was found merely in stone martens and pine martens. None of the rodent-associated Hepatozoon spp. were detected in the ticks, suggesting the possible implication of other arthropod species or non-vectorial routes in the transmission cycle of the hemoprotozoans in rodents. Our findings of H. canis in ticks other than R. sanguineus add to the observation that other ticks are also involved in the life cycle of Hepatozoon . Now that presence of Hepatozoon has been demonstrated in red foxes, gray wolves, mustelids and rodents from the Netherlands and/or Belgium, veterinary clinicians should be aware of the possibility of spill-over to domestic animals, such as dogs. Graphical Abstract

Background Hookworms (Ancylostomatidae) significantly impact on the health of both domestic animals and humans worldwide, with some species capable of causing zoonotic diseases. While hookworm infections in pets are frequently reported in Europe primarily through coproscopic studies, there are limited data regarding their presence in wild carnivores. To address this, this study aimed to assess the diversity, prevalence, and distribution of hookworms in wild canids and felids from Romania through both morphological and molecular analyses. Methods From November 2011 to February 2025, 319 carcasses belonging to six species of wild canids and felids from Romania [23 gray wolves ( Canis lupus ), 137 golden jackals ( Canis aureus ), 79 red foxes ( Vulpes vulpes ), 2 raccoon dogs ( Nyctereutes procyonoides) , 70 European wildcats ( Felis silvestris ), and 8 Eurasian lynxes ( Lynx lynx )] were collected as road kills or legally hunted. Hookworms were recovered from the intestinal tract during necropsy and preserved in formalin for morphological examination and in absolute ethanol for genetic analysis. Genomic DNA was extracted and analyzed using a PCR targeting a barcode region of the second nuclear ribosomal internal transcribed spacer (ITS-2), followed by sequencing. Sequencing results were compared with other entries from GenBank™. Results The overall hookworm infection rate was 14.1%, with hookworms detected in 4 wolves (17.4%), 23 golden jackals (16.8%), 11 European wildcats (15.7%), 4 red foxes (5.1%), 2 raccoon dogs (100%), and 1 lynx (12.5%). Three hookworm species were identified: Uncinaria stenocephala , Ancylostoma caninum , and A. tubaeforme . Molecular analysis revealed 14 unique sequences, comprising nine haplotypes of U. stenocephala , three of A. caninum , and two of A. tubaeforme . We report for the first time the Eurasian lynx as a host for A. caninum , expanding the known host range of this species. Conclusions This study provides the first comprehensive molecular assessment of hookworm diversity in European wild carnivores, showing new host–parasite associations and highlighting the importance of these hosts as reservoirs for domestic pets and, potentially, humans. The detected haplotypes showed high similarity to isolates from Europe, Asia, and the Americas, indicating a broad global connectivity of hookworm populations. Graphical abstract

Background Dirofilariosis is a parasitic mosquito-borne disease caused by members of the genus Dirofilaria , which includes Dirofilaria immitis and Dirofilaria repens. Surveillance studies in Europe have revealed that D. immitis can also be detected in a range of wild carnivores, raising questions over the impact of infections on wild carnivore animal health but also whether these populations may act as a reservoir for infection of other species, including domestic dogs and humans. Methods In the current study, we conducted surveillance for the presence of D. immitis in several wild carnivore species, including golden jackals ( Canis aureus ; n  = 77), red foxes ( Vulpes vulpes ; n  = 326), and European badgers ( Meles meles; n  = 28), collected during an annual rabies surveillance and control program from across continental and coastal regions of Croatia. Results Macroscopic examination of the right chambers of the heart during a post-mortem examination resulted in the detection of filarial parasites in 6.5% (5/77) golden jackal carcasses. Morphological identification, confirmed by molecular screening, classified all parasites as D. immitis . No D. immitis were detected in the red foxes or European badgers examined. All infected golden jackals were adults aged from 2 to 7 years with a parasite load ranging from 2 to 7 nematodes per carcass. One animal was infected with a sexually mature pair, while a second harbored pre-mature parasites; the remaining positive jackals were infected with female parasites only. Notably, histological examination of cardiac and lung tissue revealed proliferative endarteritis in the jackal with the highest parasite burden. Conclusions Further studies are required to establish whether golden jackals, as well as other wild carnivore hosts, may serve as competent definitive hosts of D. immitis and act as reservoirs for infection of other species including domestic dogs and humans. Histological changes in the cardiac tissue of at least one positive jackal were suggestive of infection with pathological consequences for the host. Graphical Abstract

Trichinella britovi is a species circulating in Iran and has already been observed in wild and domestic carnivores. The present study aims to determine the occurrence of Trichinella species in Iran's most important provinces, including the northern, northeastern, and northwest regions. In total, 93 carcasses of golden jackals, collected due to road accidents between 2019 and 2022, have been obtained from Ardabil, Mazandaran, Golestan, North Khorasan, and Khorasan Razavi provinces, Iran. Genomic DNA (four positives out of 93 carcasses) from each Trichinella genotype was amplified in a multiplex polymerase chain reaction (PCR) reaction. To confirm the multiplex PCR results, the sequences of the PCR products with cytochrome oxidase subunit I were sent to a reference laboratory within the European Union. Bioinformatics software was used to verify the sequencing results of primer fragments and to compare and analyze the results obtained with the sequences stored in the NCBI genetic database. Following multiplex PCR amplification, a two-band pattern was observed. Trichinella britovi larvae displayed the expected band pattern of 127 bp and 253 bp. The PCR products amplified using primers specific for the cytochrome oxidase subunit I gene showed 861 base pair bands in all four samples analyzed. The obtained sequences of cytochrome oxidase subunit I from all four individual muscle larvae had 100% identical haplotypes except for one nucleotide transversion in one sequence. As a result of this study, valuable insights into T. britovi prevalence and phylogenetic distribution among Iranian golden jackals have been gained in terms of prevalence and phylogenic distribution.

Pneumocystis spp. proliferate under immunosuppressive conditions in mammalian lungs, and several pathogens have been discussed as potential contributors to fungal proliferation. This study aimed to investigate the possible associations between Pneumocystis spp. and immunosuppressive viruses in Serbian wild mammals. A total of 108 wild carnivores – including golden jackals ( Canis aureus ), Eurasian badgers ( Meles meles ), and red foxes ( Vulpes vulpes ) – were collected from Veliko Gradište, Stara Pazova, and Ugrinovci during the 2022/2023 hunting season. The presence of Pneumocystis spp., canine parvovirus 2 (CPV-2), pseudorabies virus (PRV), canine distemper virus, canine coronavirus, and canine herpesvirus was assessed using conventional PCR and real-time PCR. Pneumocystis spp. were detected in 40.7% of all sampled animals (20/60 golden jackals, 4/9 Eurasian badgers, and 20/39 red foxes). CPV-2 was detected in three golden jackals (5.0% of tested golden jackals, 2.8% of all sampled animals), while PRV was found in two golden jackals (3.3% of tested golden jackals) and three red foxes (7.7% of tested red foxes; overall 4.6%). Co-infections of Pneumocystis spp. and PRV were identified in one golden jackal and two red foxes, while Pneumocystis spp. were absent in CPV-2-positive animals. All samples tested negative for other viral pathogens. No significant differences in the pathogens’ presence were observed between age groups, sexes, or sampling locations. The mean threshold cycle (Ct) values were 33.6 for Pneumocystis spp., 24.0 for CPV-2, and 31.3 for PRV. While the CPV-2 and PRV viral loads were high in co-infected samples, Pneumocystis spp. loads were associated only with subclinical infections. These findings suggest that the examined viral pathogens were unlikely to play a significant role in the development of clinically apparent Pneumocystis pneumonia, despite their potential to modulate or impair immune function. However, given the low viral prevalence and the lack of histopathological evaluation, a potential contribution of viral immunomodulation cannot be completely excluded.