Skip to main content
Download PDF

Diversity of Hepatozoon species in wild mammals and ticks in Europe

Parasites & Vectors volume 16, Article number: 27 (2023) Cite this article

This article has been updated

Abstract

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

Hepatozoon spp. (Adeleorina: Hepatozoidae) are apicomplexan vector-borne blood parasites with a complex life cycle [1, 2]. Vertebrates serve as intermediate hosts, and hematophagous arthropods such as ticks, mites and fleas serve as both definitive hosts and vectors [3,4,5,6]. Unlike other vector-borne pathogens, Hepatozoon transmission is not achieved by arthropod blood feeding on the vertebrate host, but with the infection taking place when the intermediate host ingests the definitive host. Also, transmission routes other than vector-borne have been described. For some Hepatozoon species, such as H. americanum (canids), H. sipedon (reptiles), H. caimani (caiman crocodiles) and H. ayorgbor (snakes), transmission can take place via predation of prey [7,8,9,10,11,12] or, in case of H. canis (canids) and Hepatozoon sp. of garter snakes (Thamnophis elegans), vertical transmission has been described [13,14,15,16].

The pathogenicity of Hepatozoon in wild animals such as canids [17, 18], felids [19] and mustelids [20] seems to be low, although in the case of co-infections with for example bacteria, severe disease manifestations may occur [21]. The importance of Hepatozoon in wild animals is mainly spillover to domestic animals. Hepatozoon infection in dogs and cats is known to cause subclinical to severe disease, which can be worsened by co-infection with, e.g., bacteria or other hemoparasites as well [2, 22,23,24,25,26,27,28,29,30,31].

Until a decade ago, histological methods prevailed in the characterization of blood parasites, with few exceptions [32,33,34,35,36]. Since around the 2010s, molecular methods have been commonly used [37], more specifically amplification of the small subunit ribosomal RNA (18S rRNA) gene fragments in case of detection and species identification of Hepatozoon infections. Several Hepatozoon species have been described in European wild and domestic mammals with these methods, with most studies focusing on carnivores [38, 39], e.g. H. silvestris and/or H. felis in European wild cats (Felis silvestris silvestris) in Bosnia and Herzegovina [19, 40], Spain [41] and Hungary [42], and in domestic cats from Italy [23, 43]. Also, H. martis has been detected in mustelids from Bosnia and Herzegovina and Croatia [44], Hungary [42] and Spain [41] and in wild cats from Spain [41, 45], H. ursi in brown bears (Ursus arctos) from Turkey [46] and Hepatozoon sp. in pine martens from Scotland, UK [20]. In wild canids such as red foxes (Vulpes vulpes) [17, 18, 41, 47, 48], gray wolves (Canis lupus) [49, 50] and golden jackals (Canis aureus) [51], prevalence of H. canis can be high. In contrast, prevalence of H. canis in domestic dogs [52, 53] and cats [39, 43] is generally (much) lower. Worldwide, only few studies related to Artiodactyla included testing for Hepatozoon, finding H. canis in camels from Saudi-Arabia [54] and in ticks collected from goats in China [55] and Romania [56] and Hepatozoon sp. in ticks collected from cattle in Pakistan [57]. To our knowledge, no information is available about Hepatozoon in wild Lagomorpha, except for a Spanish study in which no Hepatozoon was detected in European hares (Lepus europaeus) [35].

In rodents, Hepatozoon has been reported in Europe in Finland, Estonia and western Russia [58], Lithuania [59], Poland [60,61,62], Hungary [63], the Czech Republic [64], Slovakia [65], Great Britain [66,67,68,69], Germany [70, 71], Austria [72], Turkey [73] and Spain [32]. In rodent-related studies in which Hepatozoon could be identified to species level, in bank voles (Myodes = Clethrionomys glareolus) H. erhardovae was often detected [58, 60, 62, 63, 65, 68, 71, 72] and, to a lesser extent, H. sciuri in red squirrels (Sciuri vulgaris) [64], H. griseisciuri in gray squirrels (Sciuris carolinensis) [69] and H. lavieri in common voles (Microtus arvalis) [61].

Data about Hepatozoon in European ticks are also scarcely reported. It has long been believed that Rhipicephalus sanguineus sensu lato is the only known vector of H. canis in Europe and of many other Hepatozoon species [48, 51, 74, 75]. Recent findings of Hepatozoon sp. in other tick species raise questions about their vectorial role. Those findings include H. canis in Ixodes ricinus, I. canisuga, I. hexagonus and Dermacentor reticulatus ticks feeding on foxes in Germany [76], in Haemaphysalis concinna ticks collected from a dog in Poland [77] and in the abovementioned ticks (I. ricinus) from goats (and also dogs, fox and cat) in Romania [56]. Also, H. canis positive questing I. ricinus ticks were found in Slovakia and the Czech Republic [65] and in a R. turanicus collected from an infected fox in Italy [78].

Here, we aim to gain more knowledge on species distribution and prevalence of Hepatozoon among wild mammals and ticks in Europe. For this, we investigated a wide range of ungulates, carnivores and small mammals and ticks collected from animals and vegetation from five European countries. Animals and ticks were tested for the presence of Hepatozoon spp. using PCR and sequencing methods.

Methods

Mammals

For this study, animal samples originating from Belgium, the Netherlands, Austria, Bosnia and Herzegovina, and Croatia were used. From each animal a piece of spleen was collected for the survey. From gray wolves (Canis lupus) one or multiple samples were collected after a complete necropsy (see Additional file 3: Table S4 for detailed information). Animals were culled during regular or sanitary shooting in a period from 2010 to 2019, and no animal was shot for the purpose of this study only. All investigated gray wolves from the Netherlands were roadkill animals. All samples were collected within the frameworks of national game management and population control programs according to national laws. Samples from other mammals besides the gray wolves from the Netherlands and Belgium were gathered for previous studies [79] and used for Hepatozoon detection in this study.

Free-living rodent adults in Croatia were captured in Sherman live traps as described in an earlier study [80]. We followed animal experimentation guidelines approved by the American Society of Mammalogists [81]. Captured live animals were anesthetised in bags containing ether-soaked cotton. Deeply anesthetized animals were killed by cervical dislocation as described in the guidelines. Dead animals were aseptically dissected, and the tissue samples for DNA extraction were frozen at − 80 °C for several days before further analysis.

Ticks

Questing ticks were collected by dragging vegetation and other environments, and ticks were collected from various animals originating from Croatia and the Netherlands. Ticks were washed and stored in 98% ethanol until further processing, after morphological identification to species level using morphological keys as described in [82, 83].

DNA extraction, amplification and sequencing

DNA from animals was extracted from 10 mg of spleen and/or other organs (gray wolves) using DNA blood and tissue kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions.

For extraction of individual ticks’ DNA NucleoSpin® DNA Insect (Macherey Nagel) was used. The forward primer HepF 5’-ATACATGAGCAAAATCTCAAC-3’ and the reverse primer HepR 5’-CTTATTATTCCATGCTGCAG-3’ were used to amplify a fragment of ~ 670 bp of the 18S rRNA gene [84].

PCR reaction mixtures of 20 µl were prepared containing 10 µl G2 GOTaq mastermix (Promega, Madison, WI, USA), 7.2 µl DNase/RNase-Free distilled water (Promega), 0.4 µl 10 pmol/µl of each primer and 2 µl of sample. Positive (DNA of H. canis confirmed with sequencing from earlier studies) and negative (water from GoTaq G2 Mastermix) controls including extraction controls were used in all amplifications. The amplification product was analyzed using capillary electrophoresis on the QIAexcel system (QIAGEN, Hilden, Germany). For the purpose of further DNA sequencing, amplified PCR product was purified using ExoSAP-IT-PCR Clean-Up Reagent, according to the manufacturer’s instructions (USB Corporation, Cleveland, OH, USA). Sequencing in both directions was performed by Macrogen Europe with the same primers used for PCRs. The sequences were assembled using the SeqMan Pro software edited with EditSeq of the Lasergene software (DNASTAR, Madison WI, USA) and compared with available sequences using BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cg) system analysis.

Phylogenetic analysis

The 18S rRNA sequences of Hepatozoon obtained in this research and the deposited sequences of other Hepatozoon species and isolates available in the GenBank® were analyzed for phylogenetic relationships. The phylogenetic tree was subjected to an unweighted pair group method with arithmetic mean (UPGMA) clustering analysis using the tree builder tool incorporated in Geneious Prime (HKY). Hepatozoon sequences generated in this study were deposited in the NCBI GenBank® database under the accession numbers MH656727-MH656732 and KT274177-KT274186.

Results

Descriptive results

For this study, 2801 mammals and 754 ticks were tested for the presence of Hepatozoon with PCR. The mammals were order Artiodactyla (n = 1233), further divided in the Families Bovidae (n = 181), Suidae (n = 289) and Cervidae (n = 763); order Carnivora (n = 865), further divided in the Families Canidae (n = 336), Ursidae (n = 79), Mustelidae (n = 446) and Procyonidae (n = 4); order Eulipotyphia (n = 1), of which only the family Erinaceidae (n = 1); order Lagomorpha (n = 171), of which only the family Leporidae (n = 171) and order Rodentia (n = 531), further divided into the families Sciuridae (n = 53), Cricetidae (n = 167) and Muridae (n = 311). Of these 2801 animals, 36 originated from Bosnia and Herzegovina, 1186 from Croatia, 208 from Austria and 1371 from Belgium/the Netherlands. From Austria and Bosnia and Herzegovina, no Eulipotyphia, Lagomorpha or Rodentia were tested, and from the Netherlands/Belgium, no Eulipotyphia and Lagomorpha. Precise numbers of each mammal species and origin are given in Table 1 (Artiodactyla), Table 2 (Carnivora), Table 3 (small mammals; Rodentia, Eulipotyphia and Lagomorpha) and Additional file 1: Table S1 (all mammals).

Table 1 Presence of Hepatozoon spp. detected in samples of Artiodactyla
Full size table
Table 2 Presence of Hepatozoon spp. detected in samples from Carnivora
Full size table
Table 3 Presence of Hepatozoon spp. detected in samples from small mammals (Rodentia, Eulipotyphia and Lagomorpha)
Full size table

In total 754 ticks of 15 tick species (Table 4) were used for this study. Of these, 287 originated from the Netherlands (all collected from animals), and the rest, 467 ticks, originated from Croatia (collected from animals n = 376, collected from the environment n = 91). Table 5 specifies the results of 38 ticks that were collected from three foxes, which tested negative for the presence of Hepatozoon DNA in their spleen samples.

Table 4 Prevalence and species of Hepatozoon in ticks collected from animals and environment (questing ticks)
Full size table
Table 5 Hepatozoon in ticks collected from negative foxes
Full size table

Prevalence of Hepatozoon in mammals

See Tables 1, 2, 3 and Additional file 1: Table S1 for Hepatozoon prevalence results of 2801 tested mammals. Overall, 370 (13.2%) mammal samples tested positive for Hepatozoon. The Carnivora showed highest prevalence (34.8%), followed by Rodentia (12.1%). Artiodactyla and Lagomorpha showed the lowest Hepatozoon prevalence (0.5% and 0.6%, respectively). Within the Artiodactyla, only chamois (Bovidae) and roe deer (Cervidae) tested positive. The only animal that was tested within the order Eulipotyphia (an European hedgehog, Erinaceus europeaus) tested positive. Within the Carnivora, the four families showed differences in prevalence: the Canidae showed the highest prevalence (49.7%), and within that family especially the golden jackals (80.8%), followed by gray wolves (over half of the tested wolves were positive) and red foxes (less than half of the tested foxes were positive). Within the Mustelidae, pine martens were most frequently infected, followed by stone martens and European polecats. In contrast, all samples of the Families Ursidae and Procyonidae of the Carnivora tested negative. Within the Rodentia, the Muridae showed much lower prevalence (3.2%) than Sciuridae (24.5%) and Cricetidae (24.6%).

Regarding origin, the mammals from Croatia (n = 1186) and the Netherlands/Belgium (n = 1371) showed higher prevalence (14.6% and 13.9%, respectively) than animals from Austria (n = 208) and Bosnia (n = 36) (2.9% and 2.8%, respectively). The chamois and roe deer (Artiodactyla) that tested positive, were all from Austria. It is interesting to point out the difference in prevalence of Hepatozoon among bank voles (Croatia 81.8% and the Netherlands/Belgium 10.5%) and wood mice (Croatia 14.6% and the Netherlands/Belgium 0.0%).

Prevalence of Hepatozoon in ticks

Overall, 31 (4.1%) of the 754 collected ticks tested positive for Hepatozoon (Table 2). Ticks of the genera Dermacentor showed the highest prevalence (16.0%), followed by Ixodes (5.3%) and Rhipicephalus (1.4%). None of the ticks of the genera Hyalomma (n = 35) and Haemaphysalis (n = 23) and none of the ticks from the Netherlands (n = 287) tested positive. Ticks that were collected from animals (29/663, 4.4%) tested positive more often than ticks collected from the environment (2/91, 2.2%).

Sequence results

Results of the 18S sequence molecular analysis (Tables 1, 2, 3, 4, Figs. 1, 2, Additional file 1: Table S1, Additional file 2: Table S2) showed that five different Hepatozoon species were detected in the tested mammals and ticks. Also, Hepatozoon that could not be further specified to species level (Hepatozoon sp.) was found. Within all tested animals, H. canis was most prevalent (6.2%; 173/2801 mammals and 3.7%; 28/754 ticks). Interestingly, H. canis isolate MH656730 was most prevalent in mammals (3.8%; 107/2801) from Austria, Bosnia, Croatia and the Netherlands/Belgium, while H. canis isolate MH656729 was most often detected in ticks (2.5%; 19/754), and isolated solely from Croatian mammals (Tables 1, 2, 5 and Fig. 1). Furthermore, H. canis MH656729 was detected within the family Canidae (golden jackals and gray wolves), Mustelidae (badger), Erinaceaidae (European hedgehog) and Leporidae (European hare), while H. canis MH656730 was detected in carnivores, Canidae (golden jackals, gray wolves and red foxes), Mustelidae (badger), but also in Artiodactyla Cervidae (roe deer). Of the 38 ticks that were collected from Hepatozoon-negative foxes, in 16 (42.1%) ticks H. canis MH656729 was detected (Table 5). In Mustelidae, H. martis (MH656728) was most prevalent (96.1%; 122/127). In Sciuridae (squirrels), only H. sciuri (MH656732) was detected. In Rodentia, Hepatozoon sp. vole isolate (MH656731) was detected in bank voles (Cricetidae) from the Netherlands and Croatia and H. ayorgbor (EF157822) in yellow necked mice and wood mice (Fig. 2). Also, in bank voles, yellow-necked mice and wood mice from Croatia and the Netherlands, Hepatozoon sp. was detected (Table 1, Table 2, Fig. 2).

Fig. 1
figure 1

Phylogenetic tree (HKY, UPGMA with Babesia canis as outgroup) of Hepatozoon sequences from Carnivora. The sequences derived in this study are in bold

Full size image
Fig. 2
figure 2

Phylogenetic tree (HKY, UPGMA with Babesia canis as outgroup) of Hepatozoon sequences from Rodentia. The sequences derived in this study are in bold

Full size image

Distribution of Hepatozoon canis in organs of gray wolves

Of the 119 (Croatia n = 114, the Netherlands n = 5) wolves of which different numbers of organs (range 1 to 9) were screened to assess the distribution of H. canis, 71 (Croatia n = 66, the Netherlands n = 5) had one or more positive organ(s) (Additional File 3; Table S4). Hepatozoon canis MH656729 was detected in organs of 50 wolves and H. canis MH656730 in organs of 19 wolves. Also, in two wolves, both H. canis isolates were detected. In most of the wolves, one (n = 18) or two (n = 34) organs were tested. Three and four organs were tested in six wolves each. Six organs were tested in four wolves, eight organs in one wolf and finally nine organs in two wolves. The organs that were most tested were spleen (n = 64), lymph node (n = 36) and skeletal muscle (n = 20). Bone marrow (n = 3) and blood (n = 1) were tested the least and brain (n = 7), myocardium (n = 10), lungs (n = 12), kidneys (n = 12) and liver (n = 13) in between.

Spleen samples tested positive most often (93.8%; 60/64), followed by samples of lungs (83.3%; 10/12), lymph nodes (75.0%; 27/36) and bone marrow (66.7%; 2/3). The seven brain samples and one blood sample tested negative. Also, myocardium (60.0%; 6/10), liver (53.9%; 7/13), kidney (41.7%; 5/12) and skeletal muscle (25.0%; 5/20) samples tested positive.

Discussion

We investigated a wide range of mammal and tick species originating from five southeastern, central and western European countries for Hepatozoon prevalence and species diversity. Hepatozoon was detected in mammals from all five countries, with differences in prevalence. Hepatozoon prevalence of mammals in Austria and Bosnia and Herzogovina was ~ 3% compared to a prevalence in Croatia and Belgium/the Netherlands of 14–15%. The main reason for this difference in prevalence seems to be sampling bias. The animal species with the highest prevalence (Canis aureus, C. lupus, Vulpes vulpes, Martes foina, M. martes and Myodes glareolus; Tables 1, 2, 3, Additional file 1: Table S1) originated mainly from Croatia and Belgium/the Netherlands. From Austria and Bosnia and Herzegovina, fewer and different animal species were sampled (mainly Artilodactyla, a few Carnivora and no Rodentia). Since Hepatozoon is known to be present in Austrian [85] and Bosnian [40, 44, 86] carnivores and in Austrian rodents [72] and is most likely also present in Bosnian rodents, including samples of Carnivora and Rodentia from those countries, it probably would have increased prevalence.

High overall Hepatozoon prevalence was found in Carnivora, especially the Canidae and Mustelidae, and in Rodentia, especially the Cricetidae and Sciuridae, which is in accordance to other studies [41, 49, 63, 70, 87].

In this study, 670-bp fragments of the 18S rRNA gene were used for phylogenetic analysis and species determination. Even though for this purpose amplifications of longer fragments [88] or next-generation sequencing of nuclear, apicoplast and mitochondrial genes [89] are currently advised, we identified five different Hepatozoon species in the mammals and ticks: H. canis [90], H. felis [91] and the more recently named H. martis [44], H. sciuri [64] and H. ayorgbor [92]. Surprisingly, H. martis was not only detected in Martes foina, M. martes and other Mustelidae, but also in Artiodactyla (R. rupicapra and C. capreolus) from Austria. Also, in Austrian roe deer (C. capreolus), H. canis was detected. Broader host specificity is known for H. martis and H. canis, although the species were detected in other Carnivora, Canidae and Mustelidae [41]. The presence of H. canis in the spleen samples of roe deer and chamois therefore represents an unexpected finding. Although prevalence was low, current detection could suggest lack of host specificity as seen in other tick-borne apicomplexans, e.g. Theileria capreoli infecting gray wolves [93].

In the 31 positive (4.1%) ticks, only Hepatozoon species that are associated with Carnivores were detected (mainly H. canis and, to a lesser extent, H. martis and H. felis). This is not surprising, since Hepatozoon transmission is known to take place by carnivores (intermediate hosts) ingesting infected ticks (definitive hosts) [2]. We found H. canis not only in R. sanguineus, but also in D. reticulatus, I. hexagonus, I. ricinus, I. canisuga and I. ventalloi, which adds to the observations that other tick species than R. sanguineus may also be definitive tick hosts in the life cycle of H. canis [76, 94]. More research is necessary to confirm vector competence and capacity of these tick species for Hepatozoon since merely detection of the parasites in a tick is insufficient to designate the tick species as a vector.

In the Rodentia that were tested in our study, two Hepatozoon species could be identified: H. sciuri (in squirrels) and H. ayorgbor (in A. flavicollis and A. sylvaticus). The Hepatozoon sp. detected in bank voles (M. glareolus) from Croatia and the Netherlands could not be identified to species level. All Hepatozoon spp. sequences of bank voles from this study were (nearly) identical to each other and to Hepatozoon sequences from voles in GenBank® (Additional file 2: Table S3). As far as we know, these Hepatozoon sp. sequences were not found in any other mammalian species. Hence, we refer to this group as Hepatozoon sp. Vole isolate.

The species that were detected in rodents were not detected in the tested ticks. This could mean that ticks are not involved in the life cycle of these Hepatozoon species or that other tick species (e.g. I. triangucileps, which feeds only on small mammals [95]) and/or tick stages are involved. Hepatozoon ayorgbor has been described in snakes, ectoparasites and rodents, even though our finding in Croatian rodents (A. flavicollis and A. sylvaticus) is the first reported in European mammals. In the life cycle of H. ayorgbor, snakes can be infected via predation of rodents, with rodents serving as paratenic or intermediate hosts and mosquitoes as definitive invertebrate hosts [9], but findings in ticks and mites [96] suggest that other arthropods may be serving as definitive hosts as well. Whether the Portuguese tick (species unknown) in which H. ayorgbor was detected and of which the sequence from GenBank® (MZ475989) was used in our sequence analysis was truly infected or merely contaminated via blood feeding on an infected host is therefore not clear. Hepatozoon ayorgbor-like sequence was detected in three spleen samples of great gerbils in northwestern China [97], sharing 98.2% similarity to H. ayorgbor in the blood and liver of a ball python (Python regius) fed with tissues of mice experimentally infected with H. ayorgbor [9, 92]. Together with our findings, this confirms that rodents play a role in the life cycle of H. ayorgbor- or H. ayorgbor-related genotypes.

Our findings of Hepatozoon in carnivores and rodents from the Netherlands and Belgium are the first reported, but our study is also the first performed regarding detection of Hepatozoon in those countries. The species that were identified in mammals from the Netherlands and Belgium were H. canis in gray wolves and red foxes, H. martis in stone martens, pine martens and European polecats, and H. sciuri in squirrels. Even though a high prevalence of Hepatozoon was found in wild carnivores from the Netherlands/Belgium, to our knowledge, in The Netherlands no autochthonous Hepatozoon spillover from wild to domestic carnivores has been reported so far. No Hepatozoon was detected in the investigated Dutch and Belgian ticks. This could be because transmission of H. canis in for example foxes takes place via ticks in fox burrows (such as I. canisuga [98]) or via vertical transmission [18], which makes spillover to dogs less likely. To detect H. canis presence in ticks from the Netherlands and Belgium, other ticks than I. hexagonus and I. ricinus collected from hedgehogs would be advisable (Table 4).

The results of the investigated gray wolves show that spleen samples are most likely to test positive in case of a positive animal, which is in accordance with other reports [13].

Conclusion

Our results show that Hepatozoon is widely present in wild mammals and ticks originating from several countries in West, Central and Southeast Europe. Presence of Hepatozoon was confirmed in ticks other than the ‘usual suspect’ R. sanguineus. Besides confirming presence of Hepatozoon in wild mammals and ticks in countries in which Hepatozoon was previously detected, presence of this tick-borne parasite in the Netherlands/Belgium was demonstrated for the first time, even though circulation in ticks could not be confirmed. Since spillover from wildlife to domestic animals in countries where Hepatozoon is endemic occurs, veterinary clinicians in the Netherlands/Belgium should be aware of the presence of this tick-borne disease.

Availability of data and materials

Hepatozoon sequences generated in this study were submitted to NCBI GenBank® under accession numbers MH656727-MH656732 and KT274177-KT274186. Data supporting the conclusions of this article are included within the article and its additional files. A limited amount of DNA from samples is available upon reasonable request.

Change history

  • 26 January 2023

    Missing Open Access funding information has been added in the Funding Note.

References

  1. Smith TG. The genus Hepatozoon (Apicomplexa: Adeleina). J Parasitol. 1996;82:565–85.

    Article  CAS  Google Scholar 

  2. Baneth G. Perspectives on canine and feline hepatozoonosis. Vet Par. 2011;181:3–11.

    Article  Google Scholar 

  3. Baneth G, Samish M, Shkap V. Life cycle of Hepatozoon canis (Apicomplexa: Adeleorina: Hepatozoidae) in the tick Rhipicephalus sanguineus and domestic dog (Canis familiaris). J Parasitol. 2007;93:283–99.

    Article  Google Scholar 

  4. Merino S, Martínez J, Masello JF, Bedolla Y, Quillfeldt P. First molecular characterization of a Hepatozoon species (Apicomplexa: Hepatozoidae) infecting birds and description of a new species infecting storm petrels (Aves: Hydrobatidae). J Parasitol. 2014;100:338–43.

    Article  Google Scholar 

  5. Valkiūnas G, Mobley K, Iezhova TA. Hepatozoon ellisgreineri n. sp. (Hepatozoidae): description of the first avian apicomplexan blood parasite inhabiting granulocytes. Parasitol Res. 2016;115:609–13.

    Article  Google Scholar 

  6. Cardoso WA, Perles L, Picelli AM, Correa JKC, André MR, Viana LA. Hepatozoon parasites (Apicomplexa: Hepatozoidae) in fish Hoplias aimara (Characiformes, Erythrinidae) from the Eastern Amazon. Brazil Parasitol Res. 2022;121:1041–6.

    Article  Google Scholar 

  7. Lainson R, Paperna I, Naiff RD. Development of Hepatozoon caimani (Carini, 1909) Pess a, De Biasi & De Souza, 1972 in the Caiman Caiman c. crocodilus, the frog Rana catesbeiana and the mosquito Culex fatigans. Mem Inst Oswaldo Cruz. 2003;98:103–13.

    Article  Google Scholar 

  8. Viana LA, Soares P, Silva JE, Paiva F, Coutinho ME. Anurans as paratenic hosts in the transmission of Hepatozoon caimani to caimans Caiman yacare and Caiman latirostris. Parasitol Res. 2012;110:883–6.

    Article  Google Scholar 

  9. Sloboda M, Kamler M, Bulantová J, Votýpka J, Modrý D. Rodents as intermediate hosts of Hepatozoon ayorgbor (Apicomplexa: Adeleina: Hepatozoidae) from the African ball python, Python regius? Folia Parasitol. 2008;55:13–6.

    Article  CAS  Google Scholar 

  10. Smith TG, Desser SS, Martin DS. The development of Hepatozoon sipedon sp. nov. (Apicomplexa: Adeleina: Hepatozoidae) in its natural host, the Northern water snake (Nerodia sipedon sipedon), in the culicine vectors Culex pipiens and C. territans, and in an intermediate host, the Northern leopard frog (Rana pipiens). Parasitol Res. 1994;80:559–68.

    Article  CAS  Google Scholar 

  11. Johnson EM, Allen KE, Panciera RJ, Ewing SA, Little SE. Experimental transmission of Hepatozoon americanum to New Zealand White rabbits (Oryctolagus cuniculus) and infectivity of cystozoites for a dog. Vet Par. 2009;164:162–6.

    Article  CAS  Google Scholar 

  12. Johnson EM, Panciera RJ, Allen KE, Sheets ME, Beal JD, Ewing SA, et al. Alternate pathway of infection with Hepatozoon americanum and the epidemiologic importance of predation. J Vet Int M / ACVIM. 2009;23:1315–8.

    Article  CAS  Google Scholar 

  13. Hodžić A, Mrowietz N, Cézanne R, Bruckschwaiger P, Punz S, Habler VE, et al. Occurrence and diversity of arthropod-transmitted pathogens in red foxes (Vulpes vulpes) in western Austria, and possible vertical (transplacental) transmission of Hepatozoon canis. Parasitology. 2018;145:335–44.

    Article  Google Scholar 

  14. Murata T, Inoue M, Tateyama S, Taura Y, Nakama S. Vertical transmission of Hepatozoon canis in dogs. J Vet Med Sci. 1993;55:867–8.

    Article  CAS  Google Scholar 

  15. Kauffman KL, Sparkman A, Bronikowski AM, Palacios MG. Vertical transmission of Hepatozoon in the Garter Snake Thamnophis elegans. J Wildl Dis. 2017;53:121–5.

    Article  Google Scholar 

  16. Schäfer I, Müller E, Nijhof AM, Aupperle-Lellbach H, Loesenbeck G, Cramer S, et al. First evidence of vertical Hepatozoon canis transmission in dogs in Europe. Parasites Vectors. 2022;15:296.

    Article  Google Scholar 

  17. Cardoso L, Cortes HC, Eyal O, Reis A, Lopes AP, Vila-Viçosa MJ, et al. Molecular and histopathological detection of Hepatozoon canis in red foxes (Vulpes vulpes) from Portugal. Parasites Vectors. 2014;7:113.

    Article  Google Scholar 

  18. Mierzejewska EJ, Dwużnik D, Koczwarska J, Stańczak Ł, Opalińska P, Krokowska-Paluszak M, et al. The red fox (Vulpes vulpes), a possible reservoir of Babesia vulpes, B. canis and Hepatozoon canis and its association with the tick Dermacentor reticulatus occurrence. Ticks Tick Borne Dis. 2021;12:101551.

    Article  Google Scholar 

  19. Hodžić A, Alić A, Prašović S, Otranto D, Baneth G, Duscher GG. Hepatozoon silvestris sp. nov.: morphological and molecular characterization of a new species of Hepatozoon (Adeleorina: Hepatozoidae) from the European wild cat (Felis silvestris silvestris). Parasitology. 2017;144:650–61.

    Article  Google Scholar 

  20. Simpson VR, Panciera RJ, Hargreaves J, McGarry JW, Scholes SF, Bown KJ, et al. Myocarditis and myositis due to infection with Hepatozoon species in pine martens (Martes martes) in Scotland. Vet Rec. 2005;156:442–6.

    Article  CAS  Google Scholar 

  21. Alić A, Šupić J, Goletić T, Rešidbegović E, Lutvikadić I, Hodžić A. A unique case of fatal coinfection caused by Leptospira spp. and Hepatozoon canis in a Red Fox Cub (Vulpes vulpes). Pathogens. 2021;11(1):11

    Article  Google Scholar 

  22. Schäfer I, Kohn B, Nijhof AM, Müller E. Molecular detection of Hepatozoon species infections in domestic cats living in Germany. J Fel Med Surgery. J Fel Med Surgery. 2022;24(10):994–1000.

    Article  Google Scholar 

  23. Grillini M, Simonato G, Tessarin C, Dotto G, Traversa D, Cassini R, et al. Cytauxzoon sp. and Hepatozoon spp. in domestic cats: a preliminary study in North-Eastern Italy. Pathogens. 2021;10(9):1214.

    Article  CAS  Google Scholar 

  24. Lloret A, Addie DD, Boucraut-Baralon C, Egberink H, Frymus T, Gruffydd-Jones T, et al. Hepatozoonosis in cats: ABCD guidelines on prevention and management. J Fel Med Surg. 2015;17:642–4.

    Article  Google Scholar 

  25. Lilliehöök I, Tvedten HW, Pettersson HK, Baneth G. Hepatozoon canis infection causing a strong monocytosis with intra-monocytic gamonts and leading to erroneous leukocyte determinations. Vet Clin Pathol. 2019;48:435–40.

    Article  Google Scholar 

  26. Kegler K, Nufer U, Alic A, Posthaus H, Olias P, Basso W. Fatal infection with emerging apicomplexan parasite Hepatozoon silvestris in a domestic cat. Parasitol Vectors. 2018;11:428.

    Article  Google Scholar 

  27. Kruzeniski SJ, Tam FM, Burgess HJ. Pathology in practice. Ehrlichia-Hepatozoon coinfection in a dog. J Am Vet Med Assoc. 2013;243:1705–7.

    Article  Google Scholar 

  28. Chhabra S, Uppal SK, Singla LD. Retrospective study of clinical and hematological aspects associated with dogs naturally infected by Hepatozoon canis in Ludhiana, Punjab, India. Asian Pac J Trop Biomed. 2013;3:483–6.

    Article  CAS  Google Scholar 

  29. Baneth G, Aroch I, Tal N, Harrus S. Hepatozoon species infection in domestic cats: a retrospective study. Vet Parasitol. 1998;79:123–33.

    Article  CAS  Google Scholar 

  30. Otranto D, Dantas-Torres F, Weigl S, Latrofa MS, Stanneck D, Decaprariis D, et al. Diagnosis of Hepatozoon canis in young dogs by cytology and PCR. Parasites Vectors. 2011;4:55.

    Article  Google Scholar 

  31. Vojta L, Mrljak V, Curković S, Zivicnjak T, Marinculić A, Beck R. Molecular epizootiology of canine hepatozoonosis in Croatia. Int J Parasitol. 2009;39:1129–36.

    Article  CAS  Google Scholar 

  32. Criado-Fornelio A, Ruas JL, Casado N, Farias NA, Soares MP, Müller G, et al. New molecular data on mammalian Hepatozoon species (Apicomplexa: Adeleorina) from Brazil and Spain. J Parasitol. 2006;92:93–4.

    Article  CAS  Google Scholar 

  33. Johnson EM, Allen KE, Panciera RJ, Ewing SA, Little SE, Reichard MV. Field survey of rodents for Hepatozoon infections in an endemic focus of American canine hepatozoonosis. Vet Parasitol. 2007;150:27–32.

    Article  Google Scholar 

  34. Soares Ferreira Rodrigues AF, Daemon E, Massard CL. Morphological and morphometrical characterization of gametocytes of Hepatozoon procyonis Richards, 1961 (Protista, Apicomplexa) from a Brazilian wild procionid Nasua nasua and Procyon cancrivorus (Carnivora, Procyonidae). Parasitol Res. 2007;100:347–50.

    Article  Google Scholar 

  35. Gimenez C, Casado N, Criado-Fornelio A, de Miguel FA, Dominguez-Peñafiel G. A molecular survey of Piroplasmida and Hepatozoon isolated from domestic and wild animals in Burgos (northern Spain). Vet Parasitol. 2009;162:147–50.

    Article  CAS  Google Scholar 

  36. Allen KE, Johnson EM, Little SE. Hepatozoon spp infections in the United States. Vet Clin N Am Small Anim Pract. 2011;41:1221–38.

    Article  Google Scholar 

  37. de Waal T. Advances in diagnosis of protozoan diseases. Vet Parasitol. 2012;189:65–74.

    Article  Google Scholar 

  38. Duscher GG, Leschnik M, Fuehrer HP, Joachim A. Wildlife reservoirs for vector-borne canine, feline and zoonotic infections in Austria. Int J Par Parasites Wildl. 2015;4:88–96.

    Article  Google Scholar 

  39. Criado-Fornelio A, Buling A, Pingret JL, Etievant M, Boucraut-Baralon C, Alongi A, et al. Hemoprotozoa of domestic animals in France: prevalence and molecular characterization. Vet Parasitol. 2009;159:73–6.

    Article  CAS  Google Scholar 

  40. Hodžić A, Alić A, Duscher GG. High diversity of blood-associated parasites and bacteria in European wild cats in Bosnia and Herzegovina: a molecular study. Ticks Tick Borne Dis. 2018;9:589–93.

    Article  Google Scholar 

  41. Ortuño M, Nachum-Biala Y, García-Bocanegra I, Resa M, Berriatua E, Baneth G. An epidemiological study in wild carnivores from Spanish Mediterranean ecosystems reveals association between Leishmania infantum, Babesia spp. and Hepatozoon spp. infection and new hosts for Hepatozoon martis, Hepatozoon canis and Sarcocystis spp. Transbound Emerg Dis. 2022;69(4):2110–25.

    Article  Google Scholar 

  42. Hornok S, Boldogh SA, Takács N, Kontschán J, Szekeres S, Sós E, et al. Molecular epidemiological study on ticks and tick-borne protozoan parasites (Apicomplexa: Cytauxzoon and Hepatozoon spp) from wild cats (Felis silvestris), Mustelidae and red squirrels (Sciurus vulgaris) in central Europe, Hungary. Parasites Vectors. 2022;15:174.

    Article  CAS  Google Scholar 

  43. Giannelli A, Latrofa MS, Nachum-Biala Y, Hodžić A, Greco G, Attanasi A, et al. Three different Hepatozoon species in domestic cats from southern Italy. Ticks Tick-borne Dis. 2017;8:721–4.

    Article  Google Scholar 

  44. Hodžić A, Alić A, Beck R, Beck A, Huber D, Otranto D, et al. Hepatozoon martis n. sp. (Adeleorina: Hepatozoidae): morphological and pathological features of a Hepatozoon species infecting martens (family Mustelidae). Ticks Tick Borne Dis. 2018;9:912–20.

    Article  Google Scholar 

  45. Barandika JF, EspÍ A, Oporto B, Del Cerro A, Barral M, Povedano I, et al. Occurrence and genetic diversity of piroplasms and other apicomplexa in wild carnivores. Parasitol Open. 2016;7(2):284–90.

    Google Scholar 

  46. Akyuz M, Kirman R, Guven E. Morphological and molecular data of Hepatozoon ursi in two brown bears (Ursus arctos) in Turkey. Folia Parasitol. 2020; 67.

  47. Criado-Fornelio A, Martín-Pérez T, Verdú-Expósito C, Reinoso-Ortiz SA, Pérez-Serrano J. Molecular epidemiology of parasitic protozoa and Ehrlichia canis in wildlife in Madrid (central Spain). Parasitol Res. 2018;117:2291–8.

    Article  Google Scholar 

  48. Mitková B, Hrazdilová K, Steinbauer V, D’Amico G, Mihalca AD, Modrý D. Autochthonous Hepatozoon infection in hunting dogs and foxes from the Czech Republic. Parasitol Res. 2016;115:4167–71.

    Article  Google Scholar 

  49. Battisti E, Zanet S, Khalili S, Trisciuoglio A, Hertel B, Ferroglio E. Molecular survey on vector-borne pathogens in alpine wild carnivorans. Front Vet Sci. 2020;7:1.

    Article  Google Scholar 

  50. Hodžić A, Georges I, Postl M, Duscher GG, Jeschke D, Szentiks CA, et al. Molecular survey of tick-borne pathogens reveals a high prevalence and low genetic variability of Hepatozoon canis in free-ranging grey wolves (Canis lupus) in Germany. Ticks Tick Borne Dis. 2020;11:101389.

    Article  Google Scholar 

  51. Farkas R, Solymosi N, Takács N, Hornyák Á, Hornok S, Nachum-Biala Y, et al. First molecular evidence of Hepatozoon canis infection in red foxes and golden jackals from Hungary. Parasites Vectors. 2014;7:303.

    Article  Google Scholar 

  52. Dordio AM, Beck R, Nunes T, Pereira da Fonseca I, Gomes J. Molecular survey of vector-borne diseases in two groups of domestic dogs from Lisbon, Portugal. Parasites Vectors. 2021;14:163.

    Article  CAS  Google Scholar 

  53. Helm CS, Samson-Himmelstjerna GV, Liesner JM, Kohn B, Müller E, Schaper R, et al. Identical 18S rRNA haplotypes of Hepatozoon canis in dogs and foxes in Brandenburg, Germany. Ticks Tick Borne Dis. 2020;11:101520.

    Article  Google Scholar 

  54. Alanazi AD, Nguyen VL, Alyousif MS, Manoj RRS, Alouffi AS, Donato R, et al. Ticks and associated pathogens in camels (Camelus dromedarius) from Riyadh Province, Saudi Arabia. Parasites Vectors. 2020;13:110.

    Article  CAS  Google Scholar 

  55. Li J, Kelly P, Guo W, Zhang J, Yang Y, Liu W, et al. Molecular detection of Rickettsia, Hepatozoon, Ehrlichia and SFTSV in goat ticks. Vet Par. 2020;20:100407.

    Google Scholar 

  56. Andersson MO, Tolf C, Tamba P, Stefanache M, Radbea G, Rubel F, et al. Babesia, Theileria, and Hepatozoon species in ticks infesting animal hosts in Romania. Parasitol Res. 2017;116:2291–7.

    Article  Google Scholar 

  57. Ghafar A, Cabezas-Cruz A, Galon C, Obregon D, Gasser RB, Moutailler S, et al. Bovine ticks harbour a diverse array of microorganisms in Pakistan. Parasites Vectors. 2020;13:1.

    Article  CAS  Google Scholar 

  58. Laakkonen J, Sukura A, Oksanen A, Henttonen H, Soveri T. Haemogregarines of the genus Hepatozoon (Apicomplexa: Adeleina) in rodents from northern Europe. Folia Parasitol. 2001;48:263–7.

    Article  CAS  Google Scholar 

  59. Baltrūnaitė L, Kitrytė N, Križanauskienė A. Blood parasites (Babesia, Hepatozoon and Trypanosoma) of rodents, Lithuania: part I. Molecular and traditional microscopy approach. Parasitol Res. 2020;119:687–94.

    Article  Google Scholar 

  60. Bajer A, Welc-Falęciak R, Bednarska M, Alsarraf M, Behnke-Borowczyk J, Siński E, et al. Long-term spatiotemporal stability and dynamic changes in the haemoparasite community of bank voles (Myodes glareolus) in NE Poland. Microb Ecol. 2014;68:196–211.

    Article  Google Scholar 

  61. Pawelczyk A, Bajer A, Behnke JM, Gilbert FS, Sinski E. Factors affecting the component community structure of haemoparasites in common voles ( Microtus arvalis) from the Mazury Lake District region of Poland. Parasitol Res. 2004;92:270–84.

    Article  CAS  Google Scholar 

  62. Bajer A, Pawelczyk A, Behnke JM, Gilbert FS, Sinski E. Factors affecting the component community structure of haemoparasites in bank voles (Clethrionomys glareolus) from the Mazury Lake District region of Poland. Parasitology. 2001;122:43–54.

    Article  Google Scholar 

  63. Rigó K, Majoros G, Szekeres S, Molnár I, Jablonszky M, Majláthová V, et al. Identification of Hepatozoon erhardovae Krampitz, 1964 from bank voles (Myodes glareolus) and fleas in Southern Hungary. Parasitol Res. 2016;115:2409–13.

    Article  Google Scholar 

  64. Modrý D, Hofmannová L, Papežík P, Majerová K, Votýpka J, Hönig V, et al. Hepatozoon in Eurasian red squirrels Sciurus vulgaris, its taxonomic identity, and phylogenetic placement. Parasitol Res. 2021;120:2989–93.

    Article  Google Scholar 

  65. Hamšíková Z, Silaghi C, Rudolf I, Venclíková K, Mahríková L, Slovák M, et al. Molecular detection and phylogenetic analysis of Hepatozoon spp. in questing Ixodes ricinus ticks and rodents from Slovakia and Czech Republic. Parasitol Res. 2016;115:3897–904.

    Article  Google Scholar 

  66. Simpson VR, Birtles RJ, Bown KJ, Panciera RJ, Butler H, Davison N. Hepatozoon species infection in wild red squirrels (Sciurus vulgaris) on the Isle of Wight. Vet Rec. 2006;159:202–5.

    Article  CAS  Google Scholar 

  67. Healing TD. Infections with blood parasites in the small British rodents Apodemus sylvaticus, Clethrionomys glareolus and Microtus agrestis. Parasitology. 1981;83:179–89.

    Article  CAS  Google Scholar 

  68. Turner CM. Seasonal and age distributions of Babesia, Hepatozoon, Trypanosoma and Grahamella species in Clethrionomys glareolus and Apodemus sylvaticus populations. Parasitology. 1986;93:279–89.

    Article  Google Scholar 

  69. Watkins BM, Nowell F. Hepatozoon griseisciuri in grey squirrels (Sciurus carolinensis): changes of blood leucocyte numbers resulting from infection. Parasitology. 2003;127:115–20.

    Article  CAS  Google Scholar 

  70. Galfsky D, Król N, Pfeffer M, Obiegala A. Long-term trends of tick-borne pathogens in regard to small mammal and tick populations from Saxony, Germany. Parasites Vectors. 2019;12:131.

    Article  Google Scholar 

  71. Walter G, Liebisch A. Studies of the ecology of some blood protozoa of wild small mammals in North Germany (author’s transl). Acta Trop. 1980;37:31–40.

    CAS  Google Scholar 

  72. Sebek Z, Sixl W, Stünzner D, Valová M, Hubálek Z, Troger H. Blood parasites of small wild mammals in Steiermark and Burgenland. Folia Parasitol. 1980;27:295–301.

    CAS  Google Scholar 

  73. Usluca S, Celebi B, Karasartova D, Gureser AS, Matur F, Oktem MA, et al. Molecular survey of Babesia microti (Aconoidasida: Piroplasmida) in Wild Rodents in Turkey. J Med Entomol. 2019;56:1605–9.

    Article  Google Scholar 

  74. Geurden T, Becskei C, Six RH, Maeder S, Latrofa MS, Otranto D, et al. Detection of tick-borne pathogens in ticks from dogs and cats in different European countries. Ticks Tick Borne Dis. 2018;9:1431–6.

    Article  Google Scholar 

  75. Tolnai Z, Sréter-Lancz Z, Sréter T. Spatial distribution of Anaplasma phagocytophilum and Hepatozoon canis in red foxes (Vulpes vulpes) in Hungary. Ticks Tick Borne Dis. 2015;6:645–8.

    Article  CAS  Google Scholar 

  76. Najm N-A, Meyer-Kayser E, Hoffmann L, Pfister K, Silaghi C. Hepatozoon canis in German red foxes (Vulpes vulpes) and their ticks: molecular characterization and the phylogenetic relationship to other Hepatozoon spp. Parasitol Res. 2014;113:2679–85.

    Article  Google Scholar 

  77. Hornok S, Tánczos B, Fernández de Mera IG, de la Fuente J, Hofmann-Lehmann R, Farkas R. High prevalence of Hepatozoon-infection among shepherd dogs in a region considered to be free of Rhipicephalus sanguineus. Vet Par. 2013;196:189–93.

    Article  CAS  Google Scholar 

  78. Giannelli A, Lia RP, Annoscia G, Buonavoglia C, Lorusso E, Dantas-Torres F, et al. Rhipicephalus turanicus, a new vector of Hepatozoon canis. Parasitology. 2017;144:730–7.

    Article  CAS  Google Scholar 

  79. Azagi T, Jaarsma RI, Docters van Leeuwen A, Fonville M, Maas M, Franssen FFJ, et al. Circulation of Babesia species and their exposure to humans through Ixodes ricinus. Pathogens. 2021;10:386.

    Article  CAS  Google Scholar 

  80. Beck R, Vojta L, Curković S, Mrljak V, Margaletić J, Habrun B. Molecular survey of Babesia microti in wild rodents in central Croatia. Vector Borne Zoon Dis. 2011;11:81–3.

    Article  Google Scholar 

  81. Sikes RS, Care tA, Mammalogists UCotASo. Guidelines of the American Society of Mammalogists for the use of wild mammals in research and education. J Mammal. 2016;2016:663–88.

    Article  Google Scholar 

  82. Arthur DR. British ticks. London: Butterworths; 1963.

    Google Scholar 

  83. Hillyard PD. Shrewsbury, U.K.: Field Studies Council; 1996.

  84. Inokuma H, Okuda M, Ohno K, Shimoda K, Onishi T. Analysis of the 18S rRNA gene sequence of a Hepatozoon detected in two Japanese dogs. Vet Par. 2002;106:265–71.

    Article  CAS  Google Scholar 

  85. Trimmel NE, Walzer C. Infectious wildlife diseases in Austria—a literature review from 1980 until 2017. Front Vet Sci. 2020;7:3.

    Article  Google Scholar 

  86. Hodžić A, Alić A, Fuehrer HP, Harl J, Wille-Piazzai W, Duscher GG. A molecular survey of vector-borne pathogens in red foxes (Vulpes vulpes) from Bosnia and Herzegovina. Parasites Vectors. 2015;8:88.

    Article  Google Scholar 

  87. Sgroi G, Iatta R, Veneziano V, Bezerra-Santos MA, Lesiczka P, Hrazdilová K, et al. Molecular survey on tick-borne pathogens and Leishmania infantum in red foxes (Vulpes vulpes) from southern Italy. Ticks Tick Borne Dis. 2021;12:101669.

    Article  Google Scholar 

  88. Modrý D, Beck R, Hrazdilová K, Baneth G. A review of methods for detection of Hepatozoon infection in carnivores and arthropod vectors. Vector Borne Zoon Dis. 2017;17:66–72.

    Article  Google Scholar 

  89. Léveillé AN, Baneth G, Barta JR. Next generation sequencing from Hepatozoon canis (Apicomplexa: Coccidia: Adeleorina): complete apicoplast genome and multiple mitochondrion-associated sequences. Int J Parasitol. 2019;49:375–87.

    Article  Google Scholar 

  90. Mathew JS, Van Den Bussche RA, Ewing SA, Malayer JR, Latha BR, Panciera RJ. Phylogenetic relationships of Hepatozoon (Apicomplexa: Adeleorina) based on molecular, morphologic, and life-cycle characters. J Parasitology. 2000;86:366–72.

    Article  CAS  Google Scholar 

  91. Baneth G, Sheiner A, Eyal O, Hahn S, Beaufils JP, Anug Y, et al. Redescription of Hepatozoon felis (Apicomplexa: Hepatozoidae) based on phylogenetic analysis, tissue and blood form morphology, and possible transplacental transmission. Parasites Vectors. 2013;6:102.

    Article  Google Scholar 

  92. Sloboda M, Kamler M, Bulantová J, Votýpka J, Modrý D. A new species of Hepatozoon (Apicomplexa: Adeleorina) from Python regius (Serpentes: Pythonidae) and its experimental transmission by a mosquito vector. J Parasitol. 2007;93:1189–98.

    Article  CAS  Google Scholar 

  93. Beck A, Huber D, Polkinghorne A, Kurilj AG, Benko V, Mrljak V, et al. The prevalence and impact of Babesia canis and Theileria sp. in free-ranging grey wolf (Canis lupus) populations in Croatia. Parasites Vectors. 2017;10:168.

    Article  Google Scholar 

  94. Gabrielli S, Kumlien S, Calderini P, Brozzi A, Iori A, Cancrini G. The first report of Hepatozoon canis identified in Vulpes vulpes and ticks from Italy. Vector Borne Zoon Dis. 2010;10:855–9.

    Article  Google Scholar 

  95. Mysterud A, Byrkjeland R, Qviller L, Viljugrein H. The generalist tick Ixodes ricinus and the specialist tick Ixodes trianguliceps on shrews and rodents in a northern forest ecosystem—a role of body size even among small hosts. Parasites Vectors. 2015;8:639.

    Article  Google Scholar 

  96. Mendoza-Roldan JA, Ribeiro SR, Castilho-Onofrio V, Marcili A, Simonato BB, Latrofa MS, et al. Molecular detection of vector-borne agents in ectoparasites and reptiles from Brazil. Ticks Tick Borne Dis. 2021;12:101585.

    Article  Google Scholar 

  97. Ji N, Chen X, Liu G, Zhao S, Tan W, Liu G, et al. Theileria, Hepatozoon and Taenia infection in great gerbils (Rhombomys opimus) in northwestern China. Int J Par Parasites Wildl. 2021;15:79–86.

    Article  Google Scholar 

  98. Karbowiak G, Stanko M, Miterpaková M, Hurníková Z, Víchová B. Ticks (Acari: Ixodidae) Parasitizing Red Foxes (Vulpes vulpes) in Slovakia and new data about subgenus Pholeoixodes occurrence. Acta Parasitol. 2020;65:636–43.

    Article  Google Scholar 

Download references

Acknowledgements

We would like to thank everyone who contributed to the collection and processing of the samples.

Funding

This research was financially supported by the Dutch Ministry of Health, Welfare and Sport (VWS) and a grant from the European Interreg North Sea Region program, as part of the NorthTick project. Open access funding provided by Linköping University.

Author information

Author notes

  1. Mathilde Uiterwijk and Relja Beck contributed equally to this work

Authors and Affiliations

  1. Centre for Monitoring of Vectors (CMV), Netherlands Institute for Vectors, Invasive plants and Plant health (NIVIP), Netherlands Food and Consumer Product Safety Authority (NVWA), Wageningen, the Netherlands

    Mathilde Uiterwijk

  2. Division of Molecular Biology, Laboratory for Molecular Plant Biology and Biotechnology, Rudjer Boskovic Institute, Zagreb, Croatia

    Lea Vojta

  3. Department of Fisheries, Apiculture, Wildlife Management and Special Zoology, Faculty of Agriculture, University of Zagreb, Zagreb, Croatia

    Nikica Šprem

  4. Department of Veterinary Pathology, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia

    Ana Beck

  5. Laboratory for Parasitology, Department for Bacteriology and Parasitology, Croatian Veterinary Institute, Zagreb, Croatia

    Daria Jurković & Relja Beck

  6. Faculty of Veterinary Medicine, Dutch Wildlife Health Centre, Utrecht University, Utrecht, the Netherlands

    Marja Kik

  7. Austrian Agency for Health & Food Safety (AGES), Vienna, Austria

    Georg G. Duscher

  8. Centre for Microbiology and Environmental System Science (CMESS), Department of Microbiology and Ecosystem Science, Division of Microbial Ecology (DoME), University of Vienna, Vienna, Austria

    Adnan Hodžić

  9. Department of Forensic and State Veterinary Medicine, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia

    Slaven Reljić

  10. Centre of Infectious Disease Control of the National Institute for Public Health and the Environment (Cib-RIVM), Bilthoven, the Netherlands

    Hein Sprong

Authors
  1. Mathilde Uiterwijk
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lea Vojta
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nikica Šprem
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ana Beck
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Daria Jurković
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Marja Kik
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Georg G. Duscher
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Adnan Hodžić
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Slaven Reljić
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Hein Sprong
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Relja Beck
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All authors read and approved the final manuscript.

Corresponding author

Correspondence to Mathilde Uiterwijk.

Ethics declarations

Ethics approval and consent to participate

All animals collected from Croatia, Austria and Bosnia and Herzegovina were killed legally during regular hunting events and under the respective hunting laws. Ethical approval number of the Ethical Committee of the Croatian Veterinary Institute: Z-VI-4-2543/14. The study was carried out according to national animal welfare regulations (the Netherlands and Belgium).

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Additional file 1: Table S1.

Overall prevalence of Hepatozoon and species in tested animal species and among countries.

Additional file 2: Table S2

. Hepatozoon ayorgbor isolates from Croatian small wild rodents compared to the python isolate EF157822. Table S3. Hepatozoon sp. isolates infecting Croatian small wild rodents compared to Spanish Myodes glareolus isolates AY600625 and AY600626.

Additional file 3: Table S4

. Distribution of H. canis in organs of gray wolves.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Uiterwijk, M., Vojta, L., Šprem, N. et al. Diversity of Hepatozoon species in wild mammals and ticks in Europe. Parasites Vectors 16, 27 (2023). https://doi.org/10.1186/s13071-022-05626-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s13071-022-05626-8

Keywords

  • Tick-borne diseases
  • Apicomplexa
  • Wildlife
  • Wild carnivores
  • Wild ungulates
  • Rodents
  • 18S ribosomal DNA

Parasites & Vectors

ISSN: 1756-3305

Contact us