Hepatozoon Species Research Articles

Genetic diversity of Hepatozoon (Apicomplexa) from domestic cats in South Africa, with a global reassessment of Hepatozoon felis diversity.

Genetic diversity within partial 18S rRNA sequences from Hepatozoon protozoan parasites from domestic cats in South Africa was assessed and compared against published data to assess global biogeographic patterns. Multiple distinct haplotypes of Hepatozoon felis were identified, as well as an unrelated Hepatozoon lineage. Hepatozoon felis genetic diversity globally is very high, indicating a likely complex of species. The recently described Hepatozoon apri from wild boars is closely related to some lineages of H. felis. Sarcocystis and Babesia parasites were also detected. Since Hepatozoon felis is apparently a species complex, potential differences between genetically distinct forms need to be assessed. The finding of an unrelated Hepatozoon indicates that felids can be infected by more species of Hepatozoon than currently known, and that trophic interactions may increase the number of Hepatozoon species found in carnivores. Genetic screening again is demonstrated to identify previously unrecognised parasites from vertebrate hosts.

First Report of a Novel Hepatozoon sp. in Giant Pandas (Ailuropoda melanoleuca).

The first report of giant pandas (Ailuropoda melanoleuca) infected with a novel Hepatozoon species is presented. An intraleukocytic parasite was detected via routine blood smear from a zoo-housed giant panda at the National Zoological Park. Ribosomal DNA sequences indicated a previously undescribed Hepatozoon species. Phylogenetic and distance analyses of the sequences placed it within its own branch, clustered with Old World species with carnivore (primarily ursid and mustelid) hosts. Retrospective and opportunistic testing of other individuals produced additional positive detections (17/23, 73.9%), demonstrating 100% prevalence (14/14) across five institutions. All animals were asymptomatic at time of sampling, and health implications for giant pandas remain unknown.

Molecular survey of Babesia vogeli and Hepatozoon species in dogs from urban area of Midwestern Brazil

In Brazil, the most important tickborne pathogens affecting dogs include Babesia vogeli, Ehrlichia canis, Anaplasma platys, Hepatozoon canis, and Mycoplasma haemocanis. Babesia spp. and Hepatozoon spp., transmitted by ixodid ticks, have been reported to naturally infect dogs and are widespread. The authors aimed to investigate the incidence of B. vogeli and Hepatozoon spp. infection using molecular methods to identify factors associated with the infection in dogs from urban areas of Cuiabá municipality, Midwestern Brazil. Polymerase chain reaction (PCR) assay revealed a prevalence of 9.36% (Confidence Interval-CI 95%; 2.72%; 6.79%) and 9.61% (CI 95%; 7.0%; 13.0%) among dogs for B. vogeli and Hepatozoon, respectively. DNA sequences obtained from 10 Hepatozoon PCR positive samples were sequenced and were identical to one another and, moreover, were 100% (541/541 base of pairs-bp) homologous to the corresponding 18S rDNA sequences of H. canis. Twenty-five dogs (6.02%) generated amplicons using PCR protocols for both organisms, indicating co-infection by these two protozoans. To the best of our knowledge, our study was the first molecular survey to consider the entire population of dogs from the study area. Moreover, young dogs (0-12 months of age), as well as animals living in walled houses?without access to the street?were more susceptible to infection with B. vogeli and H. canis, respectively.

Epidemiology of tick borne pathogens of dogs in Nigerian communities

Tick borne diseases (TBDs) have significant impact on the health and welfare of domestic animals and humans. There is extremely little data on the prevalence of tick species, TBDs or their impact in Nigeria. Nigeria’s scenario is further worsened by lack of basic diagnostic facilities and treatment, compared with the average person’s income. The multiple climate zones and animal husbandry practices in Nigeria also make it difficult to extrapolate studies from one zone to the other five geopolitical zones. TBDs reported in Nigerian dogs include Anaplasma species. (A. platys A. omatnenne), Babesia species (B. rossi, B. canis and B. gibsoni), Theileria species. (T. equi, T. sable), Ehrlichia sp. (E. canis, E. ruminantum), Hepatozoon species (H. canis) and Candidatus Neoehrlichia mikurensis. The prevalence of zoonotic pathogens of dogs such as Borrelia sp., in humans in West Africa also indicates that these are likely to present in dogs in Nigeria. This main study aim is to identify ticks taken from dogs in Nigeria using morphological and molecular methods, determine which host species the ticks have fed on, and identify pathogens that they are carrying. It also aims to compare molecular and point of care diagnostics for TBDs in blood from Nigerian dogs. The overall objective is to provide robust data on which tick species and TBDs are present in Nigerian dogs and the potential zoonotic or epizootic risk.

Assessing the natural circulation of canine vector-borne pathogens in foxes, ticks and fleas in protected areas of Argentine Patagonia with negligible dog participation.

We collected blood and/or ectoparasites from 49 South American grey foxes (Lycalopex griseus) and two Andean foxes (L. culpaeus) caught in two National Parks of southern Argentine Patagonia (Bosques Petrificados, BPNP; and Monte León, MLNP) where dogs are nearly absent (density < 0.01 dog/km2). Common ectoparasites were the flea Pulex irritans (88% prevalence) and the tick Amblyomma tigrinum (29%). Conventional PCR and sequencing of 49 blood samples, 299 fleas analysed in 78 pools, and 21 ticks revealed the presence of DNA of the following canine vector-borne pathogens: in grey foxes, Rickettsia sp. (3%), hemoplasmas (8%), including Mycoplasma haemocanis, and Hepatozoon sp. (50%); in P. irritans, Bartonella spp. (72% of flea pools from 76% of foxes), mostly B. vinsonii subsp. berkhoffii but also B. rochalimae, Anaplasmataceae (Wolbachia sp.; 60% and 54%), and M. haemocanis/haemofelis (29% and 18%); and in A. tigrinum, Hepatozoon sp. (33% of ticks in 4 of 7 foxes). No piroplasmid DNA was detected in any sample. Andean foxes were negative for all tested pathogens. Two different Hepatozoon haplotypes were detected: the most prevalent was phylogenetically associated with H. felis, and the other with H. americanum and related sequences. Amblyomma tigrinum and Hepatozoon sp. were more abundant and/or prevalent in BPNP than in colder MLNP, 300 km southwards, perhaps located close to the limit for tick suitability. Bartonella v. berkhoffii was also significantly more prevalent in fleas of foxes in BPNP than in MLNP. This study provides novel information about natural host-pathogen associations in wildlife, markedly extends the distribution area in South America of arthropods and vector-borne pathogens of veterinary and public health interest, and contributes preliminary evidence about the potential role of A. tigrinum and P. irritans as vectors, respectively, for potentially new species of Hepatozoon from Lycalopex spp. and for M. haemocanis that should be further investigated.

A Molecular Study on Hepatozoon canis Infection in Dogs in Tehran (Iran).

Hepatozoonosis is a protozoal disease caused by various species of Hepatozoon. This parasite is transmitted from tick; the main vector of Hepatozoon canis is usually the brown dog tick (Rhipicephalus sanguineus). However, several species of ticks are disposed as the alternative vectors. Dogs are usually infected by eating the tick or a part of the tick organ infected by the mature oocysts containing infectious sporozoite. In the current study, a total of 145 blood samples were collected from the cephalic vein of pet, stray, and shelter dogs in Tehran. To conduct this study, first thin blood smears were prepared from all the samples and stained with the Giemsa method. Then, after extraction of DNA from the blood samples, in order to trace Hepatozoon canis, the 18S rRNA gene segment of the parasite was amplified using polymerase chain reaction (PCR). To confirm the PCR-positive results, five randomly selected PCR-positive samples were sequenced. According to the results, through direct observation of microscopic slides, no infection of H. canis parasite was observed, but according to the PCR results, 32 out of the 145 blood samples were found to be infected by H. canis. In this study, infection to H. canis in older dogs was higher than in young dogs, and more male dogs were found to be infected by the parasite compared to female dogs; but no significant difference was observed in this regard (P &amp;gt; 0.05). Moreover, stray dogs showed a significantly higher rate of infection, compared to the pet and shelter ones (P &amp;lt; 0.05).

Hepatozoon milleri sp. nov. (Adeleorina: Hepatozoidae) in Akodon montensis (Rodentia: Cricetidae: Sigmodontinae) from southeastern Brazil.

Members of the genus Hepatozoon (Miller, 1908) are blood parasites found in a wide range of host species, including wild rodents; however, information about the life cycle, distribution and Hepatozoon species diversity infecting these mammals are lacking. We studied the parasite stages and DNA sequences of Hepatozoon sp. of 11 naturally infected Akodon montensis. Thin blood smears, tissue samples and whole blood were obtained for morphology, morphometry and molecular analyses. Seven of the 11 rodents had gamonts on the blood smears. Biological and morphological features of the parasite such as tissue tropism, gamonts and meronts size and morphology, as well as the DNA sequence comparison and phylogenetic analysis, indicated that the Hepatozoon sp. detected in this study is distinct from those species previously reported in small rodents. Herein, we propose a new species, named Hepatozoon milleri sp. nov. This is the first description of a new Hepatozoon species from wild small rodents in Brazil, based on morphological and molecular characteristics.

Molecular characterization and identification of Hepatozoon species Miller, 1908 (Apicomplexa: Adeleina: Hepatozoidae) in captive snakes from Brazil.

Species of Hepatozoon are parasites frequently recorded in snakes. The species identification of this genus was based mostly on the gametocyte morphology and morphometric calculations. For more reliable results, molecular characterization, an initial step for the correct identification of Hepatozoon species, has been used. This study aimed to determine the prevalence and identification of Hepatozoon species in captive snakes from Brazil. To that end, morphological, morphometric, and molecular data were obtained. A total of 157 snakes; 128 venomous (Crotalus durissus) and 29 non-venomous (Epicrates crassus and Boa constrictor) were screened for Hepatozoon blood parasites. Using light microscopy, 20 (12.78%) snakes were found positive for Hepatozoon spp., of which 6/29 (20.7%) were non-venomous and 14/128 (10.9%) were venomous; all with low parasitemia. Polymerase chain reaction (PCR), performed with the primers HepF300/Hep900, confirmed all 20 (100%) samples positive for hemogregarines. Species of Hepatozoon were identified from eight sequenced samples. Two previously described species, Hepatozoon cuestensis and Hepatozoon musa, were identified. The present study is the first to report H. musa within the snake hosts E. crassus and C. durrisus. In addition, a potentially new Hepatozoon species from B. constrictor was identified.

Fatal infection with emerging apicomplexan parasite Hepatozoon silvestris in a domestic cat

BackgroundHepatozoon silvestris is an emerging apicomplexan parasite discovered in European wild cats from Bosnia and Herzegovina and blood samples of a domestic cat from Southern Italy in 2017. It has also been identified in Ixodes ricinus collected from a domestic cat in Wales, UK, in 2018. The clinical relevance, pathogenesis and epidemiology of this novel Hepatozoon species are not yet understood. Thus, the objective of this paper was to report and describe the first fatal case of an H. silvestris infection in a domestic cat.ResultsThe cat, which originated from Switzerland, died shortly after presenting clinical signs of lethargy, weakness and anorexia. At necropsy, no specific lesions were observed. Histopathology of the heart revealed a severe lympho-plasmacytic and histiocytic myocarditis. Mature and developing protozoal meronts morphologically compatible with Hepatozoon species were observed associated with the myocardial inflammation. No other lesions were present in any other organ evaluated, and the cat tested negative for retroviral and other immunosuppressive infectious agents. Polymerase chain reaction from the myocardium resulted in a specific amplicon of the Hepatozoon 18S rRNA gene. Sequencing and BLAST analysis revealed 100% sequence identity with H. silvestris.ConclusionsThe severity of the infection with fatal outcome in an otherwise healthy animal suggests a high virulence of H. silvestris for domestic cats. The presence of this emerging parasite in a domestic cat in Switzerland with no travel history provides further evidence for a geographical distribution throughout Europe.

Molecular characterisation of Hepatozoon aegypti Bashtar, Boulos & Mehlhorn, 1984 parasitising the blood of Spalerosophis diadema (Serpentes: Colubridae).

Hepatozoon aegypti Bashtar, Boulos & Mehlhorn, 1984 was first described from the blood of the diadem snake (Spalerosophis diadema) in Egypt. During an investigation of the diversity of reptilian haemogregarines in Saudi Arabia, seven diadem snakes (100% of the sample) were found to be highly parasitised by H. aegypti, with an average parasitaemia of 37% per 500 counted erythrocytes. A complete characterisation of this species with morphometrics and 18S rDNA sequence data is therefore presented here. The infection was found to be restricted to the erythrocytes with, frequently, single and, sometimes, double infections. Mature gamonts were sausage-shaped with round posterior and anterior extremities and measured 14 (13-17) × 3.5 (3-5) μm. The infected erythrocytes were hypertrophied with a faintly stained cytoplasm and longitudinally stretched nuclei. The merogonic stages occurred only in the endothelial cells of the snakes' lungs, and no stages were found in other organs. Mature meronts were round in shape, measured 18 (17-21) μm in diameter and were estimated to produce between 9 and 15 merozoites. Phylogenetic analysis based on the partial 18S small subunit ribosomal DNA sequences indicates that Hepatozoon aegypti cluster within a mixed clade of Hepatozoon species parasitising snakes, geckos and rodents from various geographic areas. Our results might reinforce the theory of prey-predator transmission in respect to the relationships of snake-host Hepatozoon species.

Two new species of Hepatozoon (Apicomplexa: Hepatozoidae) parasitising species of Philothamnus (Ophidia: Colubridae) from South Africa.

To date, only a few species of Hepatozoon Miller, 1908 have been described from amphibians and reptiles of South Africa, including two species from anuran hosts, three from saurians, one from chelonians, and two from ophidians. Hepatozoon bitis (Fantham, 1925) and Hepatozoon refringens (Sambon et Seligmann, 1907), parasitising Bitis arientans (Merrem) and Pseudoaspis cana (Linnaeus), respectively, were described in the early 1900s and since then there have been no further species of Hepatozoon described from snakes in South Africa. Blood smears, used in peripheral blood haemogregarine stage morphometrics, and whole blood used in molecular characterisation of haemogregarines were collected from the caudal vein of six snakes of three species, namely Philothamnus hoplogaster (Günther), Philothamnus semivariegatus (Smith) and Philothamnus natalensis natalensis (Smith). For comparison, a comprehensive table summarising available information on species of Hepatozoon from African snakes is presented. Haemogregarines found infecting the snakes from the present study were morphologically and molecularly different from any previously described from Africa and are thus here described as Hepatozoon angeladaviesae sp. n. and Hepatozoon cecilhoarei sp. n. Both haemogregarine species were observed to cause considerable dehaemoglobinisation of the host cell, in case of infection with H. angeladaviesae resulting in a characteristic peripheral undulation of the host cell membrane and karyorrhexis. To the authors' knowledge, these are the first haemogregarines parasitising snakes of the genus Philothamnus Smith described using both morphological and molecular characteristics in Africa.

Hepatozoon martis n. sp. (Adeleorina: Hepatozoidae): Morphological and pathological features of a Hepatozoon species infecting martens (family Mustelidae)

Species of the genus Hepatozoon (Adeleorina: Hepatozoidae) are arthropod-transmitted protozoan parasites that infect a wide range of vertebrate hosts. In the present study, we describe a new species of Hepatozoon primarily infecting martens and propose the name Hepatozoon martis n. sp., based on its unique morphological, molecular and pathogenic features. The overall prevalence of infection with H. martis n. sp. assessed by PCR in European pine martens (Martes martes) from Bosnia and Herzegovina and stone martens (Martes foina) from Croatia was 100% and 64%, respectively. Gamonts were found in neutrophils and monocytes, and various developmental stages were described in tissue cross-sections. Hepatozoon martis n. sp. shows a high predilection for muscle tissue, and the heart was the most frequently affected organ among the tissues tested by histopathology. Microscopically, pyogranulomatous lesions associated with the presence of the parasitic forms were observed in the cardiac and skeletal muscles of all positive animals examined. Furthermore, the possible existence of alternative, non-vectorial routes of transmission is discussed.

Endemic, exotic and novel apicomplexan parasites detected during a national study of ticks from companion animals in Australia

BackgroundApicomplexan tick-borne pathogens that cause disease in companion animals include species of Babesia Starcovici, 1893, Cytauxzoon Neitz & Thomas, 1948, Hepatozoon Miller, 1908 and Theileria Bettencourt, Franca & Borges, 1907. The only apicomplexan tick-borne disease of companion animals that is known to occur in Australia is babesiosis, caused by Babesia canis vogeli Reichenow, 1937 and Babesia gibsoni Patton, 1910. However, no molecular investigations have widely investigated members of Apicomplexa Levine, 1980 in Australian ticks that parasitise dogs, cats or horses, until this present investigation.ResultsTicks (n = 711) removed from dogs (n = 498), cats (n = 139) and horses (n = 74) throughout Australia were screened for piroplasms and Hepatozoon spp. using conventional PCR and Sanger sequencing. The tick-borne pathogen B. vogeli was identified in two Rhipicephalus sanguineus Latreille ticks from dogs residing in the Northern Territory and Queensland (QLD). Theileria orientalis Yakimov & Sudachenkov, 1931 genotype Ikeda was detected in three Haemaphysalis longicornis Neumann ticks from dogs in New South Wales. Unexpectedly, the exotic tick-borne pathogen Hepatozoon canis James, 1905 was identified in an Ixodes holocyclus Neumann tick from a dog in QLD. Eight novel piroplasm and Hepatozoon species were identified and described in native ticks and named as follows: Babesia lohae n. sp., Babesia mackerrasorum n. sp., Hepatozoon banethi n. sp., Hepatozoon ewingi n. sp., Theileria apogeana n. sp., Theileria palmeri n. sp., Theileria paparinii n. sp. and Theileria worthingtonorum n. sp. Additionally, a novel cf. Sarcocystidae sp. sequence was obtained from Ixodes tasmani Neumann but could not be confidently identified at the genus level.ConclusionsNovel species of parasites in ticks represent an unknown threat to the health of companion animals that are bitten by these native tick species. The vector potential of Australian ticks for the newly discovered apicomplexans needs to be assessed, and further clinical and molecular investigations of these parasites, particularly in blood samples from dogs, cats and horses, is required to determine their potential for pathogenicity.

Canine hepatozoonosis: an update

Canine hepatozoonosis is caused by two different Hepatozoon species, responsible for distinct clinical syndromes. Hepatozoonosis beyond the United States is caused by Hepatozoon canis, transmitted by the tick Rhipicephalus sanguineus, whereas North American hepatozoonosis is caused by H. americanum, transmitted by Amblyomma maculatum. The former appears to be well adapted to its canine host, causing mostly subclinical or a mild clinical disease. Anorexia, depression, lethargy, fever, lymphadenopathy and body-weight loss are the most common clinical manifestations, which may be precipitated by concurrent infectious diseases or drug-induced immunosuppression. H. canis infection is mosdy diagnosed by the detection of intraneutrophilic gamonts in Giemsa-stained blood smears. Occasionally, a high level parasitemia may be observed (100% of circulating neutrophils), though moderate parasitemia (1-5%) is more common. Imidocarb dipropionate, with or without doxycycline, usually leads in rapid clinical remission and elimination of parasitemia, although relapses are not uncommon. In marked contrast, H. americanum, a recendy emerged pathogen, apparendy poorly adapted to dogs, is responsible for a severe chronic debilitating and often fatal disease. Fever, generalized pain, muscle atrophy and reluctance to move are prominent clinical symptoms attributable mainly to severe pyogranulomatous myositis and periosteal proliferative lesions. Since parasitemia is rarely found in H. americanum infection, a solid diagnosis should be based on muscle histopathology and/or serology. The combination of trimethoprime-sulphadiazine, clindamycin and pyrimethamine (induction phase), followed by long-term administration of the anticoccidial decoquinate (maintenance phase), achieves clinical remission and prolongs survival time, although relapses may frequently occur.

Monophyly of the species of Hepatozoon (Adeleorina: Hepatozoidae) parasitizing (African) anurans, with the description of three new species from hyperoliid frogs in South Africa.

Haemogregarines (Apicomplexa: Adeleiorina) are a diverse group of haemoparasites reported from almost all vertebrate classes. The most commonly recorded haemogregarines to parasitize anurans are species of Hepatozoon Miller, 1908. To date 16 Hepatozoon species have been described from anurans in Africa, with only a single species, Hepatozoon hyperolli (Hoare, 1932), infecting a member of the Hyperoliidae. Furthermore, only two Hepatozoon species are known from South African anurans, namely Hepatozoon theileri (Laveran, 1905) and Hepatozoon ixoxo Netherlands, Cook and Smit, 2014, from Amietia delalandii (syn. Amietia quecketti) and three Sclerophrys species, respectively. Blood samples were collected from a total of 225 individuals representing nine hyperoliid species from several localities throughout northern KwaZulu-Natal, South Africa. Twenty frogs from three species were found positive for haemogregarines, namely Afrixalus fornasinii (6/14), Hyperolius argus (2/39), and Hyperolius marmoratus (12/74). Based on morphological characteristics, morphometrics and molecular findings three new haemogregarine species, Hepatozoon involucrum Netherlands, Cook and Smit n. sp., Hepatozoon tenuis Netherlands, Cook and Smit n. sp. and Hepatozoon thori Netherlands, Cook and Smit n. sp., are described from hyperoliid hosts. Furthermore, molecular analyses show anuran Hepatozoon species to be a separate monophyletic group, with species isolated from African hosts forming a monophyletic clade within this cluster.

Genetic diversity of Hepatozoon spp. in Hydrochoerus hydrochaeris and Pecari tajacu from eastern Amazon

This study aimed to identify and characterize genetically species of the genus Hepatozoon detected in Hydrochoerus hydrochaeris (capybaras) and Pecari tajacu (collared peccaries) from two localities from the Eastern Amazon. Blood samples from 196 free-living H. hydrochaeris from Marajó Island and 109 P. tajacu kept in captivity in Belém, Pará, were collected and analyzed for the presence of Hepatozoon spp. Partial sequences of the 18S rRNA gene were obtained and analyzed in comparison to others available in the NCBI database. Our results demonstrated a high prevalence of Hepatozoon canis in both mammals and the existence of four haplotypes of Hepatozoon spp., three of Hepatozoon canis and one of Hepatozoon cuestensis, found only in H. hydrochaeris. In addition, these data increase the genetic diversity of H. canis from the Eastern Amazon, as well as reporting, for the first time, the infection of mammals by H. cuestensis and P. tajacu by H. canis.

Molecular confirmation of Hepatozoon canis in Mauritius

In this study, Hepatozoon species was molecularly identified and characterized for the first time on the Indian Ocean island of Mauritius. Partial sequences of the 18S rRNA gene of the Hepatozoon isolates were analysed from three naturally infected dogs. The sequences of H. canis were similar to the 18S rRNA partial sequences (JX112783, AB365071 99%) from dog blood samples from West Indies and Nigeria. Our sequences were deposited in the GenBank database.

Hepatozoon caimani in Caiman crocodilus yacare (Crocodylia, Alligatoridae) from North Pantanal, Brazil.

Hepatozoon species are the most common intracellular hemoparasite found in reptiles. Hepatozoon caimani, whose vectors are Culex mosquitoes, has been detected in a high prevalence among caimans in Brazil by blood smears examinations. The present work aimed to detect and characterize the Hepatozoon spp. found in 33 caimans (24 free-ranging and 9 captive; 28 males and 5 females) (Caiman crocodilus yacare) sampled at Poconé, North Pantanal, state of Mato Grosso, Brazil, using blood smears examinations and molecular techniques. Hepatozoon spp.-gametocytes were found in 70.8% (17/24) and 88.8% (8/9) of blood smears from free-ranging and captive caimans, respectively. Hepatozoon spp. 18S rRNA DNA was found in 79.2% (19/24) and 88.8% (8/9) of free-ranging and captive caimans, respectively. Comparative analysis of parasitized and non-parasitized erythrocytes showed that all analyzed features were significantly different (P<0.05) for both linear and area dimensions. Phylogenetic analysis based on 18S rRNA sequences grouped the Hepatozoon spp. sequences detected in the present study together with H. caimani, recently detected in caimans in southern Pantanal.

Didelphis albiventris naturally infected with Hepatozoon canis in southeastern Brazil

Hepatozoon species are vector-borne pathogens that infect domestic and wild animals. Marsupials of the species Didelphis albiventris are adapted to urban and peri-urban areas and act as reservoir hosts for several parasites. The present study evaluated the occurrence of infection by Hepatozoon species in synantropic D. albiventris from Botucatu, São Paulo, Brazil. Blood samples and ectoparasites from 19 D. albiventris were collected from urban and peri-urban areas. Hepatozoon spp. detection was performed by microscopy and molecular analysis. One opossum was positive for Hepatozoon spp. in microscopy analysis and PCR, while another animal was positive only in PCR. The obtained sequences were 100% identical to Hepatozoon canis. Six species of ticks and two species of fleas were detected on D. albiventris. This is the first report of H. canis in synantropic D. albiventris. In Brazil, H. canis transmission among dog populations is not well established, which highlights the importance of investigating the role that opossums might play in the epidemiology of this protozoan.

Babesia, Theileria, and Hepatozoon species in ticks infesting animal hosts in Romania.

Babesia spp., Theileria spp., and Hepatozoon spp. are tick-transmitted apicomplexan parasites that cause several important diseases in animals. To increase current knowledge about the diversity of tick-transmitted pathogens in Romania, we investigated the occurrence of Babesia spp., Theileria spp., and Hepatozoon spp. in a wide range of tick species infesting animal hosts. We collected 852 ticks from 10 different animal species from 20 counties in Romania. The assessment was based on detection of parasite DNA by PCR. Five different apicomplexan parasite species were detected; among them three different species of Babesia: B. canis, B. microti, and B. ovis. Hepatozoon canis was the most frequently detected parasite, found predominately in Ixodes ricinus ticks collected from domestic dogs. It was also detected in I. ricinus collected from goat, fox, and cat. Furthermore, H. canis was found in Haemaphysalis punctata and Haemaphysalis concinna ticks. In addition, Theileria buffeli was detected in Rhipicephalus bursa ticks collected from cattle.