Hepatozoon Species Research Articles

Molecular confirmation of high prevalence of species of Hepatozoon infection in free-ranging African wild dogs (Lycaon pictus) in the Kruger National Park, South Africa.

Reports in the literature indicate that species of Hepatozoon commonly occur in African wild dog (AWD) or painted wolf (Lycaon pictus) populations. These findings were based on examination of blood smears by microscopy, and specific identity of the Hepatozoon sp. gamonts seen could not be confirmed. We present the first in-depth molecular data on the prevalence of species of Hepatozoon in a free-ranging AWD population. In a general health survey of AWDs in the Kruger National Park, blood specimens (n = 74) collected from 54 individuals were examined for the presence of Hepatozoon spp. At first sampling, specimens from 42 of 54 individuals (77.7%) were positive, based on the primer set HepF300 and HepR900. Twenty individuals were resampled between 51 and 69 days after first sampling; one of these was resampled twice. Samples from six individuals that had tested negative previously now reacted positive. Assuming that all 54 individuals were still alive, the prevalence had therefore increased to 48 individuals infected, or 88.8%. Resultant 18S rDNA sequences isolated from these specimens share high similarity to other Hepatozoon canis genotypes. Phylogenetic analysis recovered the Hepatozoon sp. isolated from AWDs within the H. canis cluster, which includes species of Hepatozoon from other canid and tick hosts.

Description of a New Species Hepatozoon quagliattus sp. nov. (Apicomplexa: Adeleorina: Hepatozoidae), infecting the Sleep Snake, Dipsas mikanii (Squamata: Colubridae: Dipsadinae) from Goiás State, Brazil.

Species of Hepatozoon Miller, 1908 (Hepatozoidae) are blood protozoans with a cosmopolitan distribution and are reported to parasitize a range of vertebrate hosts including mammals, birds, reptiles, and amphibians. The present study aimed to describe a new species of Hepatozoon (Apicomplexa: Adeleorina: Hepatozoidae) found infecting the sleep snake Dipsas mikanii (Schlegel, 1837) (Squamata: Colubridae: Dipsadinae). The snake was collected in 2017 at the municipality of Britânia, Goiás State, Brazil. Blood smears were made in order to find blood gametocytes and PCR was performed targeting the 18S rRNA gene. Microscopy screening of blood smears revealed the presence of intraerythrocytic gamont stages of Hepatozoon sp. in the peripheral blood with a parasitemia of 0.25%. Furthermore, meronts and monozoic cysts were observed in histological sections of the liver from the infected individual. The interspecific divergence of 18S rRNA sequences fragments isolated from D. mikanii had differences (2.39-11.3%) as compared to other sequences of species of Hepatozoon from snakes. Based on morphological and molecular data, a new species of Hepatozoon infecting D. mikanii from Brazil is described.

New insights on the diversity of Brazilian anuran blood parasites: With the description of three new species of Hepatozoon (Apicomplexa: Hepatozoidae) from Leptodactylidae anurans.

The Amphibia are considered the most threatened vertebrate class globally, yet in Brazil they are also one of the more diverse and species rich groups. Although, in recent years there has been strong focus on amphibian related research, their parasites have not received the same attention. In Brazil, only a single species of Hepatozoon, namely H. leptodactyli (Lesage, 1908) Pessoa, 1970, has been described from anuran hosts. The present study aimed to describe three new species of Hepatozoon parasitising Leptodactylus labyrinthicus and Leptodactylus latrans from Mato Grosso State, Brazil. From 66 anurans screened for haemogregarines, four belonging to the Leptodactylidae were found positive for species of Hepatozoon. Based on the morphological analysis of peripheral blood gamonts and spleen and liver tissue meronts, three different morphotypes of Hepatozoon spp. were identified. Morphotype 1 (M1) and morphotype 2 (M2) in L. labyrinthicus and morphotype 3 (M3) in L. latrans. Molecular data based on partial 18S rDNA sequences revealed an interspecific divergence, between the species ranging from 0.43% to 1.16%. Phylogenetic analysis recovered isolates from the present study monophyletic with other isolates from Brazilian reptile and anuran hosts, sister to a clade comprising species isolated from African, North American and European reptile and anuran host species. Thus, using morphological and molecular analysis three new species infecting Brazilian Leptodactylidae anurans were identified and described. This study increases the knowledge of Brazilian anurans blood parasites and demonstrates the importance of using integrative approaches for diagnosis of hemoparasites.

Molecular and morphological description of the first Hepatozoon (Apicomplexa: Hepatozoidae) species infecting a neotropical turtle, with an approach to its phylogenetic relationships.

Haemogregarines (Adeleorina) have a high prevalence in turtles. Nevertheless, there is only one Hepatozoon species described that infects Testudines so far; it is Hepatozoon fitzsimonsi which infects the African tortoise Kinixys belliana. Colombia harbours a great diversity of chelonians; however, most of them are threatened. It is important to identify and characterize chelonian haemoparasite infections to improve the clinical assessments, treatments and the conservation and reintroduction programs of these animals. To evaluate such infections for the Colombian wood turtle Rhinoclemmys melanosterna, we analysed blood from 70 individuals. By using the morphological characteristics of blood stages as well as molecular information (18S rRNA sequences), here we report a new Hepatozoon species that represents the first report of a hepatozoid species infecting a semi-aquatic continental turtle in the world. Although the isolated lineage clusters within the phylogenetic clades that have morphological species of parasites already determined, their low nodal support makes their position within each group inconclusive. It is important to identify new molecular markers to improve parasite species identification. In-depth research on blood parasites infecting turtles is essential for increasing knowledge that could assess this potential unknown threat, to inform the conservation of turtles and for increasing the state of knowledge on parasites.

Haemoprotozoan surveillance in peri-urban native and introduced wildlife from Australia

Vector-borne haemoprotozoans comprise a diverse group of eukaryote single-celled organisms transmitted by haematophagous (blood-feeding) invertebrates. They can cause debilitating diseases that impact wildlife, livestock, companion animals and humans. Recent research has shown that Australian wildlife host a diverse range of haemoprotozoan species; however, to date this work has primarily been confined to a few host species or isolated populations in rural habitats. There has been little investigation into the presence of these blood parasites in wildlife inhabiting urban and peri-urban areas. In this study, blood and tissue samples and ticks were collected from wildlife in New South Wales and Western Australia. Extracted DNA samples were screened with pan-specific molecular assays to determine the presence of haemoprotozoans using amplicon metabarcoding and Sanger sequencing approaches. In addition, light microscopy was performed on blood films. Eight haemoprotozoans were identified in the present study, which included species of Babesia, Hepatozoon, Theileria and Trypanosoma. Blood samples were collected from 134 animals; 70 black rats (Rattus), 18 common brush-tailed possums (Trichosurus vulpecula), two bush rats (Rattus fuscipes), 22 chuditch (Dasyurus geoffroii), 20 long-nosed bandicoots (Perameles nasuta), one quenda (Isoodon fusciventer) and one swamp rat (Rattus lutreolus). Molecular screening of DNA extracted from blood samples identified 52.2% (95% CI: 43.8–60.5%) of individuals were positive for at least one haemoprotozoan species, with 19.4% (95% CI: 13.4–26.7%) positive for more than one species. The present study provides the first sequences of Theileria cf. peramelis from black rats and long-nosed bandicoots. Babesia lohae was identified from brush-tailed possums. Two Hepatozoon genotypes were identified from black rats and bush rats. Black rats showed the highest haemoprotozoan diversity, with five species identified. No known human pathogens that have been described in the northern hemisphere were identified in the present study, and future work is required to understand the zoonotic potential of these microbes in Australia. This work represents the first large-scale body of research using molecular tools to investigate haemoprotozoans in animals at the urban-wildland interface. Further research is needed to investigate potential consequences of infection in wildlife, particularly effects of pathogen spillover from invasive black rats to native wildlife.

Genetic diversity of Hepatozoon spp. in rodents from Chile.

This study aimed to investigate the genetic diversity of Hepatozoon spp. in rodents from Valdivia, Chile. A total of 74 rodents (synanthropic n=38; wild n=36) were trapped in Valdivia. We performed conventional PCR assays for Apicomplexa organisms targeting two overlapping 18S rDNA gene fragments (600 bp and 900 bp) followed by sequencing of selected amplicons. Hepatozoon spp. occurrence was 82.43% (61/74). Twelve sequences obtained from the 600 bp and ten from the 900 bp 18S rDNA fragments were identified as Hepatozoon sp. Six sequences obtained from 18S rDNA-based overlapping PCR protocols were used for concatenated (1,400 bp) phylogenetic, haplotype and distance analyses. Hepatozoon spp. 18S rDNA concatenated sequences from the present study were detected in Oligoryzomys longicaudatus, Rattus norvegicus, Mus musculus, and Abrothrix longipilis grouped with Hepatozoon species earlier described in rodents and reptiles from Chile and Brazil. Nucleotide polymorphism of the six 18S rDNA sequences (1,400 bp) from this study, and other Chilean sequences from rodents and rodent's ticks, showed high diversity with a total of nine Chilean haplotypes. Three haplotypes from Valdivia were identified for the first time in this study, suggesting the circulation of novel haplotypes in rodents from southern Chile.

Morphological and molecular data of Hepatozoon ursi in two brown bears (Ursus arctos) in Turkey.

Species of Hepatozoon Miller, 1908 are vector-borne parasites that infect domestic and wild animals worldwide. Hepatozoon ursi Kubo, Uni, Agatsuma, Nagataki, Panciera et al., 2008 was reported from bears (Ursidae) in Japan and India. The present study represents the first report of infection with H. ursi in Turkish brown bears (Ursus arctos Linnaeus) by microscopic and molecular analysis. Two dead brown bears were found in Uzundere and Pasinler districts of Erzurum. Blood and visceral organ (spleen and liver) samples were delivered to laboratory by the Nature Conservation and National Parks officers. Detected gamonts were evaluated based on morphological features and confirmed as gamonts of H. ursi. The size of gamonts and parasitemia were 8.2 × 3.5 μm (6.9-8.7 × 3.0-3.9 μm; n = 12) and 0.6% (6/1000 leukocytes), respectively. The blood and visceral organ samples were positive for species of Hepatozoon by PCR targeting partial sequence of 18S rDNA. Sequence analysis of newly obtained sequences of H. ursi showed 98.8-100% identity with previously sequenced isolates of H. ursi. Sequences of H. ursi from Erzurum were identical to each other and showed 100% identity with isolates of H. ursi from ticks Ixodes ricinus (Linnaeus), Rhipicephalus turanicus Pomerantzev and Hyalomma marginatum Koch collected from two brown bears in Turkey (GenBank accession numbers MN463021, MN463022, MN905023). Analysis of partial sequences of the 18S rRNA gene of H. ursi showed that Turkish isolates differ in NT substitutions found at three different positions [72 (A→G), 537 (A→G) and 570 (A→T)]. This study provides morphological and molecular data of H. ursi infection in brown bears from two districts of Erzurum, Turkey. Further studies are needed to elucidate whether brown bears have any eco-epidemiologic importance in the life cycle of H. ursi in wildlife.

Morphometric and molecular characterisation of Hepatozoon bashtari n. sp. in painted saw-scaled viper, Echis coloratus (Ophidia, Viperidae).

Hepatozoon species are the most widely known haemogregarines infecting a wide range of vertebrates, although predominately snakes. Herein, Hepatozoon bashtari n. sp., originally infecting the painted saw-scaled viper, Echis coloratus, in Saudi Arabia is described using both morphological features and molecular data from 18S rDNA sequences. The overall prevalence of infection was 60% (9/15) with parasitaemia ranging from 52 to 60%. Gamonts were entirely intraerythrocytic and were observed to cause considerable hypertrophy within the host cell. The mean size of mature gamonts was 15.4 × 3.3 μm. Merogonic stages were confined to the lung endothelial cells with monomorphic meronts. The average size of mature meronts was 32 × 12 μm and they were estimated to produce 13-16 merozoites each. The phylogenetic tree generated from SSU rDNA sequences revealed that Hepatozoon bashtari sp. n. clusters with the vast majority of other Hepatozoon species infecting snakes, lizards and geckos in various regions of the world, which would appear to support the hypothesis of prey-predator transmission of the genus Hepatozoon. Through a combination of morphological comparison with closely related Hepatozoon spp. and 18S rRNA gene sequence analysis, it is possible to confirm Hepatozoon bashtari sp. n. as a new species.

Prevalence and distribution of pathogen infection and permethrin resistance in tropical and temperate populations of Rhipicephalus sanguineus s.l. collected worldwide.

The brown dog tick, Rhipicephalus sanguineus sensu lato (s.l.) Latreille (Acari: Ixodidae), is a peridomestic, cosmopolitan parasite of dogs known to vector numerous pathogens of veterinary and medical importance. Recent phylogenetic analyses separate this tick into temperate and tropical lineages. Populations of Rh. sanguineus s.l. have been reported to exhibit sodium channel target site insensitivity to permethrin and etofenprox, which is likely due to the prolonged use of pyrethroids against many pests in and around the home. In this study, populations collected in the Caribbean, Africa, Asia, Europe and North America, were tested to identify the distribution of a known resistance mechanism, pathogen-vector interactions and phylogeny in relation to latitude. Using molecular assays, populations from 29 distinct locations were simultaneously geographically typed and screened for bacterial infection by Rickettsia, Ehrlichia, Babesia and Hepatozoon species, and for the presence of a sodium channel single nucleotide polymorphism known to confer permethrin resistance. Implications of these results on Rh. sanguineus s.l. management in association with geographical distribution will be discussed.

Molecular detection of Apicomplexan hemoparasites in anurans from Brazil

Amphibians are among the most threatened vertebrate groups in the world, and the main causes include climate change, habitat destruction, and emerging diseases. Herein, we investigated the occurrence and characterized molecularly Apicomplexa in anurans from southeastern Brazil. Forty individuals from seven anuran species were sampled in São Paulo state. In the molecular analyses, one Leptodactylus latrans and one Rhinella diptycha were positive in PCR assays for species of Hepatozoon. Two L. latrans were also positive for coccidian infections (Lankesterella sp. and an unidentified coccidian species). Phylogenetic analysis based on 18S rDNA clustered the sequences detected in anurans from the present study with Hepatozoon spp. detected in reptiles and other anurans from Brazil, albeit they were separate from Hepatozoon haplotypes detected in frogs from Africa and North America. Our study showed, for the first time, the molecular detection of Lankesterella sp. and another coccidian in L. latrans. Additionally, co-infection by different species of Hepatozoon haplotypes and an unidentified coccidian in anurans from Brazil was documented.

Molecular identification of ticks and tick borne pathogens in Nigerian Dogs: A public health threat

Global warming has changed the range of tick species and the tick borne diseases (TBDs) they carry; there is an increasing need to assess the risk and impact of TBDs. This study focused on TBDs of Dogs in Nigeria, where despite a large burden of ticks and TBDs there is little data on the prevalence of either for veterinarians and human health professionals to make informed decisions regarding treatment and control. Most existing studies on cattle have indicated that Anaplasma, Babesia, Theileria, Ehrlichia, Hepatozoon and Candidatus Neoehrlichia species are present in Nigeria. However, dogs kept in much closer contact with their owners, may represent a higher risk of zoonotic TBDs transmission. This study used a combination of morphological and molecular identification of ticks and tick borne pathogens, and IDEXX 4Dx kits for pathogen antibody detection from canine blood, we examined 93 dogs to date. Rhipicephalus sanguineus was the most abundant tick found on Nigerian dogs. Babesia species were the most abundant tick borne pathogen, followed by Ehrlichia species. This study serves as the first report of Babesia microti like species, Theileria parva, Babesia bovis, Anaplasma phagocytophilium and Ehrlichia chaffeensis in Nigerian dogs. There was no evidence of Borrelia Burgdorferi sensu lato nor Dirofilaria species in Nigerian dogs. The presence of Babesia microti and Anaplasma phagocytophilium is a direct public threat. Further testing of additional 200 dog samples and multivariate modelling of demographic risk factors, will enable us develop treatment and control guidelines for TBDs for Nigerian Veterinarians and Dog owners.

Molecular detection of Hepatozoon canis in dogs and ticks in Shaanxi province, China

Hepatozoon canis, transmitted by Rhipicephalus sanguineus, is a tick-borne pathogen and causes canine hepatozoonosis. Until now, only limited previous studies were conducted on the molecular detection and characterization of Hepatozoon sp. in dogs in China. Blood samples were collected from 93 sick dogs that were clinically diagnosed as babesiosis but tested negative for Babesia, and 103 apparently healthy dogs, as well as their infesting ticks in Xi’an and Hanzhong cities, Shaanxi province of China. PCR amplifying partial 18S rRNA gene was used to detect the DNA of Hepatozoon sp. Genetic and phylogenetic analysis were performed to determine the Hepatozoon species. Our results demonstrated that H. canis was identified from the sick dogs and the infested ticks in Hanzhong, with no significant differences of prevalence between both genders and ages. No positive blood or tick samples were found in Xi’an. Moreover, all the 18S rRNA gene sequences recovered from both dogs and the infested ticks showed a high genetic similarity with each other, and also presented a close relationship with other known sequences in and outside China. In conclusion, H. canis was identified in babesiosis-suspected dogs and ticks infesting them in Shaanxi, China, although the association between clinical signs and H. canis need further study.

Redescription of Hepatozoon ameivae (Carini and Rudolph, 1912) from the lizard Ameiva ameiva (Linnaeus, 1758).

Blood samples from 72 Ameiva ameiva lizards from Central Amazonian upland forests were collected, and thin smears of 40 (55.5%) animals were positive for gamonts of Hepatozoon with a mean level of intensity of infection of 14 parasites/2000 blood erythrocytes (0.73%). The gametocytes were found attached with host cells' nuclei, and their dimensions were 14.28 ± 1.05μm in length and 4.50 ± 0.80μm in width. Phylogenetic analyses of the 18S rRNA gene showed that the new sequences obtained from A. ameiva constitute a monophyletic sister clade to the Hepatozoon spp. from Brazilian snakes. Based on morphological features and new molecular data, we redescribe this hemogregarine as Hepatozoon ameivae. This study also provides the first molecular characterization of a Hepatozoon species from a Brazilian lizard.

Molecular characterisation and morphological description of two new species of Hepatozoon Miller, 1908 (Apicomplexa: Adeleorina: Hepatozoidae) infecting leukocytes of African leopards Panthera pardus pardus (L.)

BackgroundThe African leopard Panthera pardus pardus (L.) is currently listed as a vulnerable species on the IUCN (International Union for the Conservation of Nature) red list of threatened species due to ongoing population declines. This implies that leopard-specific parasites are also vulnerable to extinction. Intracellular apicomplexan haemoparasites from the genus Hepatozoon Miller, 1908 have been widely reported from wild carnivores in Africa, including non-specific reports from leopards. This paper describes two new haemogregarines in captive and wild leopards from South Africa and provides a tabular summary of these species in relation to species of Hepatozoon reported from mammalian carnivores.MethodsBlood was collected from nine captive and eight wild leopards at various localities throughout South Africa. Thin blood smears were Giemsa-stained and screened for intraleukocytic haemoparasites. Gamont stages were micrographed and morphometrically compared with existing literature pertaining to infections in felid hosts. Haemogregarine specific primer set 4558F and 2733R was used to target the 18S rRNA gene for molecular analysis. Resulting sequences were compared to each other and with other available representative mammalian carnivore Hepatozoon sequences from GenBank.ResultsTwo species of Hepatozoon were found in captive and wild leopards. Of the 17 leopards screened, eight were infected with one or both morphologically and genetically distinct haemogregarines. When compared with other species of Hepatozoon reported from felids, the two species from this study were morphometrically and molecularly distinct. Species of Hepatozoon from this study were observed to exclusively parasitize a particular type of leukocyte, with Hepatozoon luiperdjie n. sp. infecting neutrophils and Hepatozoon ingwe n. sp. infecting lymphocytes. Phylogenetic analysis showed that these haemogregarines are genetically distinct, with Hepatozoon luiperdjie n. sp. and Hepatozoon ingwe n. sp. falling in well supported separate clades.ConclusionsTo our knowledge, this is the first morphometric and molecular description of Hepatozoon in captive and wild African leopards in South Africa. This study highlights the value of using both morphometric and molecular characteristics when describing species of Hepatozoon from felid hosts.

BASELINE HEALTH AND DISEASE ASSESSMENT OF FOUNDER EASTERN QUOLLS (DASYURUS VIVERRINUS) DURING A CONSERVATION TRANSLOCATION TO MAINLAND AUSTRALIA.

We evaluated the health of 31 (eight males, 23 females) founder eastern quolls (Dasyurus viverrinus), translocated to a fenced reserve in the Australian Capital Territory between February 2016 and July 2017. Quolls were wild caught in Tasmania (16 animals) or captive bred at Mount Rothwell Biodiversity Interpretation Centre, Victoria (15 animals). Quolls were assessed for the presence of selected potential pathogens (Toxoplasma gondii, herpesviruses, Salmonella serovars, hemoprotozoa, and ectoparasites). We assessed the relationships among sex, provenance (captive or free ranging), T. gondii or herpesvirus infection, weight, and hematologic and biochemical variables. Six of 21 quolls (29%) tested were seropositive for antibodies to T. gondii. Seropositive quolls weighed significantly more and had significantly lower potassium levels, anion gaps, and urea and triglyceride levels than seronegative quolls had. Eighteen of 31 (58%) combined conjunctival-pharyngeal-cloacal swabs collected from quolls were PCR positive for a newly identified gammaherpesvirus, tentatively named dasyurid gammaherpesvirus 3. There were no significant differences among hematologic and biochemical variables or body weights from PCR-positive and PCR-negative quolls. Eighteen of 18 (100%) of rectal-swab samples were culture negative for Salmonella serovars. Three species of tick (Ixodes tasmani, Ixodes fecialis, and Ixodes holocyclus), two species of mite (Andreacus radfordi, one unidentified), and four species of flea (Pygiopsylla hoplia, Acanthopsylla rothschildi rothschildi, Uropsylla tasmanica, and Stephanocircus dasyuri), were detected on wild-caught quolls, whereas a fifth species of flea, Echidnophaga myremecobii, was detected only on captive-bred quolls. Five of 15 blood samples (33%) were positive for hemoprotozoan DNA via PCR, a novel Hepatozoon species, a novel Theileria species, Theileria paparinii, and Trypanosoma copemani were detected. Despite the presence of several potential pathogens known to be associated with disease in other marsupials, the quolls were considered to be in good general health, suitable for translocation, and a viable population was subsequently established.

Turkey tick news: A molecular investigation into the presence of tick-borne pathogens in host-seeking ticks in Anatolia; Initial evidence of putative vectors and pathogens, and footsteps of a secretly rising vector tick, Haemaphysalis parva

This Turkey-based study investigated the presence of various tick-borne microorganisms in a broad-range of host-seeking ticks (n = 1019) that exhibit both hunter and ambusher characteristics. All collected ticks were analyzed individually via PCR-sequencing, resulting in the identification of 18 different microorganisms: six Babesia spp., including one putative novel species (Ba. occultans, Ba. crassa, Ba. rossi, Babesia sp. tavsan1, Babesia sp. tavsan2, and Babesia sp. nov.); six SFG rickettsiae (Ri. aeschlimannii, Ri. s. mongolitimonae, Ri. slovaca, Ri. raoultii, Ri. monacensis, and Ri. hoogstraalii); two Borrelia burgdorferi sensu lato spp. (Bo. afzelii and Bo. lusitaniae); two unnamed Hepatozoon spp.; Theileria annulata; and Hemolivia mauritanica. This provided evidence for the natural transstadial survival of these tick-borne microorganisms in adult ticks (in addition a nymph) of Turkey. Surprisingly, this study determined the presence of five different microorganisms (Ba. crassa, Ba. rossi, Babesia sp. Ucbas, Hepatozoon sp., and Ri. hoogstraalii) in host-seeking Haemaphysalis parva adults, for which poor data exist on its vectorial competence. Therefore, this study provides important data indicating the potential vectorial capacity of Ha. parva. This study also revealed the presence of the close ecological and evolutionary relationships between two important vector ticks, Hyalomma marginatum and Hy. aegyptium and determined genetic variations (distinct phylogenetic divergences inside the main clades) in some pathogenic SFG rickettsiae that are found in these ticks. Additionally, the presence of two Babesia species described very recently in hares with unknown vectors, namely Babesia sp. tavsan1 and Babesia sp. tavsan2, were detected for the first time in ticks. Finally, two unnamed Hepatozoon spp. were detected in Haemaphysalis ticks and their phylogenetic positions were demonstrated. Consequently, this study provides important data on the diversity of tick-borne microorganisms in host-seeking ticks and on potentially novel microorganisms (Babesia and Hepatozoon species) and their possible vectors (Ha. parva, Ha. sulcata, Hy. aegyptium, Hy. marginatum, and Rh. turanicus).

Multilocus sequencing of Hepatozoon cf. griseisciuri infections in Ontario eastern gray squirrels (Sciurus carolinensis) uncovers two genotypically distinct sympatric parasite species.

Intra-leukocytic gamonts consistent with the description of Hepatozoon griseisciuri Clark, 1958 are reported for the first time in Canadian eastern gray squirrels (Sciurus carolinensis Gmelin, 1788). Polymerase chain reaction (PCR) amplification and direct Sanger sequencing identified a pair of distinct genotypes at both a nuclear and mitochondrial locus; two 18S ribosomal RNA gene sequences (rDNA; genotype A and genotype B: 1816 base pairs (bp); 98.8% pairwise identity) and 2 distinct complete mitochondrial genome sequences (genotype A: 6311bp; genotype B: 6114bp; 89.1% pairwise identity) were obtained from 3 H. griseisciuri-infected squirrels sampled in Guelph, Ontario. The genetic content of both circular-mapping mitochondrial genomes was conventional for apicomplexan protists; each encoded for 3 protein-coding genes (cytochrome c oxidase subunit I (COI); cytochrome c oxidase subunit III (COIII); and cytochrome B (CytB)), 14 fragmented large subunit rDNA, 10 fragmented small subunit rDNA, and 8 unassigned rDNA. These genotypes, based on sequences obtained from a pair of loci from two parasite genomes, confirm the presence of at least two Hepatozoon species infecting Ontario eastern gray squirrels, one of which is likely to be conspecific with H. griseisciuri.

Quba rayonunun dağ zonalarının Qafqaz kərtənkələsinin (Darevskia Caucasusica) qan parazitlərinin (Hepatozoon, Adeleida) ilk kəşfi

The genus Hepatozoon Miller, 1908 (Apicomplexa: Adeleorina) is composed of intracellular haemogregarine parasites that are widely distributed among all tetrapod groups. The present study is microscopic data on haemogregarine parasites from the Caucasian lizard (Darevskia caucasica) of the mountain zones of the Guba district. We examined blood smears for the presence of species of Hepatozoon from three Caucasian lizards. The investigation revealed the first finding of blood parasites (Hepatozoon, Adeleida) for the Caucasian lizard of Azerbaijan, while possible vectors and pathogenicity are still largely unknown.

Molecular identification and phylogenetic analysis of Cryptosporidium, Hepatozoon and Spirometra in snakes from central China.

Snakes are popular as food and traditional medicine in China. However, information about parasitic and bacterial infections in snakes from China is scarce. We investigated the prevalence of selected zoonotic agents including Cryptosporidium, Hepatozoon and Spirometra, in snakes in central China from June to October in 2018 by PCR amplification using parasite-specific primers. PCR amplification and DNA sequencing showed that 10.1% (15/149) of snakes were positive for Cryptosporidium spp., while 2.7% (4/149) were positive for Hepatozoon. Additionally, we found 36.9% (55/149) of snakes were infected with Spirometra erinaceieuropaei. The spargana burden per infected snake ranged from 1 to 26. BLAST and phylogenetic analysis of small subunit ribosomal RNA (SSU rRNA) gene and 60-kDa glycoprotein (gp60) gene showed that the parasites belonged to Cryptosporidium parvum genotype IIdA15G1, C. baileyi, C. serpentis and a Hepatozoon species. We conclude that intensively farmed snakes excrete C. parvum and C. baileyi oocysts due to ingestion of infected feeder animals, and that wild snakes in central China were commonly infected with S. erinaceieuropaei, suggesting that eating improperly cooked snakes could be risky to human health.

Genetic diversity of Hepatozoon spp. in rodents from Brazil

Hepatozoon spp. are Apicomplexan protozoa that parasitize a wide diversity of vertebrate hosts. In Brazil, few studies have reported the occurrence of Hepatozoon spp. in rodent species. Additionally, an evaluation of the population structure and distribution of Hepatozoon species over several Brazilian biomes has not yet been performed. The present work aimed to investigate the genetic diversity of Hepatozoon spp. in rodents from 31 genera sampled in five Brazilian biomes. Samples were submitted to PCR assays for Hepatozoon spp. targeting two regions of the 18S rRNA gene. Infection by Hepatozoon spp. was detected in 195 (42.2%) rodents comprising 24 genera. Phylogenetic analyses of 18S rRNA sequences grouped all sequences in the clade of Hepatozoon spp. previously detected in rodents and reptiles, apart from those detected in domestic/wild carnivores. These data raise two non-exclusive hypotheses: (i) rodents play an important role as intermediate or paratenic hosts for Hepatozoon infections in reptiles; and (ii) rodents do not seem to participate in the epidemiology of Hepatozoon infections of domestic/wild canids and felids in Brazil. TCS analyses performed with available 18S rRNA Hepatozoon sequences detected in rodents from Brazil showed the occurrence of six haplotypes, which were distributed in two large groups: one from rodents inhabiting the coastal region of Brazil and Mato Grosso state, and another from rodents from the central region of the country. A wide survey of the South American territory will help to elucidate the evolutionary history of Hepatozoon spp. parasitizing Rodentia in the American continent.