The analyses of molecular data represent an effective tool for increasing the credibility of taxonomy and facilitate the description of species. Nevertheless, in haemoprotozoa, the growing amount of available sequential data is not matched by the still limited number of well-defined species. We identified four protistan haemoparasites in North Iranian reptiles: two Hepatozoon and two Schellackia species. Hepatozoon colubri and Hepatozoon ophisauri were morphologically identified in their type hosts, their partial 18S rDNA was analyzed, and thorough literature data were included in their redescription. The scarce data on the detected Schellackia spp. did not allow for their formal species description. Using an integrative approach, including morphological and geographical features, host specificity, molecular data, and the data published thus far, we face the following main difficulties hindering reliable diagnosis. (1) The lack of molecular data on well-described and named species. (2) The insufficiency of using only morphological and biological features, or only sequential data without morphology, to perform an absolutely reliable species diagnosis. (3) Typical morphological features are more substantial than metric means. (4) High risk of synonymy is present in taxonomy of blood Protista. (5) Artefacts caused by blood smear processing further complicate the correct morphological determination.
- MeSH
- druhová specificita MeSH
- Eucoccidiida klasifikace genetika MeSH
- kokcidióza parazitologie MeSH
- plazi parazitologie MeSH
- protozoální DNA genetika MeSH
- RNA ribozomální 18S genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Írán MeSH
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.
- MeSH
- Eucoccidiida * genetika izolace a purifikace MeSH
- fylogeneze MeSH
- játra parazitologie MeSH
- klíšťata parazitologie MeSH
- klíště parazitologie MeSH
- krev parazitologie MeSH
- medvědovití * parazitologie MeSH
- RNA ribozomální 18S genetika MeSH
- slezina parazitologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Turecko MeSH
Apicomplexan haemoparasites of the genera Schellackia Reichenow, 1919, and Karyolysus Labbé, 1894, seem to be common in lizards and widespread across the world. For decades, their identification has been based on morphological descriptions and life cycle patterns, with molecular characterizations, applied only recently. We used molecular characterization to confirm the identification of haemoparasites detected by microscopy in blood smears of Lacerta schreiberi Bedriaga, 1878, a lizard of the Iberian Peninsula. Since blood samples other than blood smears were not available from the studied lizards, 264 engorged ticks Ixodes ricinus (Linneaus, 1758) collected from them were used as an alternative non-invasive source of haemoparasite DNA for molecular genetic analyses. Of the 48 blood smears microscopically examined, 31 were positive for blood parasites (64.6% prevalence). We identified trophozoites and gamonts similar to Karyolysus lacazei (Labbé, 1894) (24/48; 50%) and Schellackia-like sporozoites (20/48; 41.7%). Mixed infections with both species occurred in 13 blood smears (27.1%). Sequence data were obtained for both parasites from engorged ticks. Phylogenetic analyses placed our unique haemogregarine sequence within the Karyolysus clade, nevertheless, within substantial polytomy. Thus, according to its morphology and effect on the host cell, we refer to this haemogregarine as Karyolysus cf. lacazei. Besides the Schellackia sequences being identical to a previously identified haplotype, we also obtained sequences of three new closely related haplotypes.
- MeSH
- Eucoccidiida klasifikace MeSH
- fylogeneze MeSH
- genetická variace * MeSH
- haplotypy MeSH
- ještěři parazitologie MeSH
- klíště parazitologie MeSH
- kokcidióza krev parazitologie veterinární MeSH
- protozoální DNA genetika MeSH
- sekvenční analýza DNA MeSH
- sporozoiti genetika MeSH
- trofozoiti genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Knowledge of blood parasites in Brazilian chelonians is limited, since they have been recorded in only six species. Mesoclemmys vanderhaegei (Bour) is a freshwater turtle with a wide geographic distribution in Brazil, but there is little information about its natural history. This paper reports on a study of the prevalence and infection intensity of a haemogregarine in two subpopulations of M. vanderhaegei. The study was conducted in two areas of Cerrado in the Upper Paraguay River basin in the state of Mato Grosso, Brazil, between November 2010 and August 2013. Ninety-five (53%) of the 179 turtles captured were positive for haemogregarine parasites. The parasitic forms observed were two morphotypes of intraerythrocytic gametocytes. The prevalence differed between size classes, increasing significantly according to the animals' body size. There was no significant difference between prevalence and sex, or between sampling periods. The mean parasite intensity was 9 parasites/2,000 erythrocytes (0.45%) and the parasite population presented an aggregated distribution, with an aggregation index of 19 and discrepancy of 0.772. This is the first record of a hemoparasite in the freshwater turtle M. vanderhaegei.
- MeSH
- Eucoccidiida fyziologie MeSH
- kokcidióza epidemiologie parazitologie veterinární MeSH
- pastviny MeSH
- prevalence MeSH
- želvy * MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Brazílie MeSH
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.
- MeSH
- Colubridae parazitologie MeSH
- Eucoccidiida klasifikace cytologie fyziologie MeSH
- interakce hostitele a parazita * MeSH
- kokcidióza parazitologie patofyziologie veterinární MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Jihoafrická republika MeSH
Vector-borne protists of the genus Hepatozoon belong to the apicomplexan suborder Adeleorina. The taxonomy of Hepatozoon is unsettled and different phylogenetic clades probably represent evolutionary units deserving the status of separate genera. Throughout our review, we focus on the monophyletic assemblage of Hepatozoon spp. from carnivores, classified as Hepatozoon sensu stricto that includes important pathogens of domestic and free-ranging canine and feline hosts. We provide an overview of diagnostic methods and approaches from classical detection in biological materials, through serological tests to nucleic acid amplification tests (NAATs). Critical review of used primers for the 18S rDNA is provided, together with information on individual primer pairs. Extension of used NAATs target to cover also mitochondrial genes is suggested as a key step in understanding the diversity and molecular epidemiology of Hepatozoon infections in mammals.
- MeSH
- Carnivora parazitologie MeSH
- členovci - vektory parazitologie MeSH
- Eucoccidiida genetika izolace a purifikace MeSH
- polymerázová řetězová reakce metody veterinární MeSH
- protozoální infekce zvířat diagnóza parazitologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Endemic Sicilian pond turtles Emys trinacris Fritz, Fattizzo, Guicking, Tripepi, Pennisi, Lenk, Joger et Wink were examined for the presence of haemogregarine parasites. The presence of haemogregarines, occurring mainly in the microgametocyte stage (13.2 ± 0.12 μm in length and 6.4 ± 0.52 μm in width), was observed in approximately 9% of the sampled E. trinacris. Based on the observed morphology and on the sequencing of nuclear 18S rDNA, we identified the parasite as Haemogregarina stepanowi Danilewsky, 1885. Morphometric study of uninfected and infected red blood cells has shown that H. stepanowi induces different changes in erythrocyte shape depending on the infective stage. The differential count of leukocytes in specimens infected with H. stepanowi showed no significant difference compared with healthy specimens. However, considering the health problems which might be induced by H. stepanowi in the closely related European pond turtle Emys orbicularis (Linneaus), monitoring of the health status of the infected Sicilian populations of E. trinacris is desirable. The restricted distribution of populations of Emys infected with haemogregarines in Sicily is quite puzzling and the possible human-mediated introduction of the parasite in Sicily is briefly discussed.
- MeSH
- Eucoccidiida klasifikace genetika izolace a purifikace fyziologie MeSH
- fylogeneze MeSH
- kokcidióza epidemiologie parazitologie veterinární MeSH
- pijavice fyziologie MeSH
- RNA ribozomální 18S genetika MeSH
- želvy parazitologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Sicilie MeSH
Blood samples from 21 red foxes (Vulpes vulpes) and 8 hunting dogs from the same locality in the Czech Republic were examined for presence of Hepatozoon canis/Hepatozoon sp. The dogs were selected based on their close contact with foxes during fox bolting and because they had not traveled into known endemic areas. Using diagnostic PCR amplifying partial 18S rDNA fragment, Hepatozoon DNA was detected in 20 red foxes (95 %) and 4 dogs (50 %). From 8 positive foxes and 2 positive dogs, we obtained nearly complete 18S rDNA sequences. Phylogenetic analyses of these sequences revealed very low variability. Buffy coat smears from positive dogs were prepared and examined. No Hepatozoon gamonts were found. This study provides the first report of autochthonous infection of H. canis/Hepatozoon in dogs and foxes from the Czech Republic. Our study indirectly demonstrates cross infection between red foxes and dogs and confirms autochthonous infection of Hepatozoon canis in dogs living in a geographic area well outside the range of Rhipicephalus sanguineus sensu lato, which is so far the only known vector of H. canis in Europe.
- MeSH
- arachnida jako vektory parazitologie MeSH
- Eucoccidiida klasifikace genetika izolace a purifikace MeSH
- fylogeneze MeSH
- kokcidióza epidemiologie parazitologie přenos veterinární MeSH
- lišky parazitologie MeSH
- nemoci psů epidemiologie parazitologie přenos MeSH
- psi MeSH
- Rhipicephalus sanguineus parazitologie MeSH
- ribozomální DNA genetika MeSH
- sekvenční analýza DNA veterinární MeSH
- sekvenční seřazení veterinární MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- psi MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika epidemiologie MeSH
By amplification and sequencing of 18S rRNA gene fragments, Hepatozoon spp. DNA was detected in 0.08 % (4/5057) and 0.04 % (1/2473) of questing Ixodes ricinus ticks from Slovakia and Czech Republic, respectively. Hepatozoon spp. DNA was also detected in spleen and/or lungs of 4.45 % (27/606) of rodents from Slovakia. Prevalence of infection was significantly higher in Myodes glareolus (11.45 %) than in Apodemus spp. (0.28 %) (P < 0.001). Sequencing of 18S rRNA Hepatozoon spp. gene amplicons from I. ricinus showed 100 % identity with Hepatozoon canis isolates from red foxes or dogs in Europe. Phylogenetic analysis showed that at least two H. canis 18S rRNA genotypes exist in Slovakia of which one was identified also in the Czech Republic. The finding of H. canis in questing I. ricinus suggests the geographical spread of the parasite and a potential role of other ticks as its vectors in areas where Rhipicephalus sanguineus is not endemic. Sequencing of 18S rRNA gene amplicons from M. glareolus revealed the presence of two closely related genetic variants, Hepatozoon sp. SK1 and Hepatozoon sp. SK2, showing 99-100 % identity with isolates from M. glareolus from other European countries. Phylogenetic analysis demonstrates that 18S rRNA variants SK1 and SK2 correspond to previously described genotypes UR1 and UR2 of H. erhardovae, respectively. The isolate from Apodemus flavicollis (Hepatozoon sp. SK3b) was 99 % identical with isolates from reptiles in Africa and Asia. Further studies are necessary to identify the taxonomic status of Hepatozoon spp. parasitizing rodents in Europe and the host-parasite interactions in natural foci.
- MeSH
- arachnida jako vektory parazitologie MeSH
- Arvicolinae parazitologie MeSH
- Eucoccidiida klasifikace genetika izolace a purifikace MeSH
- fylogeneze MeSH
- klíště parazitologie MeSH
- kokcidióza epidemiologie parazitologie MeSH
- lidé MeSH
- Murinae parazitologie MeSH
- ribozomální DNA chemie genetika MeSH
- sekvenční analýza DNA MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika epidemiologie MeSH
- Slovenská republika epidemiologie MeSH
The majority of Haemogregarina species have been based on the morphology of their erythrocytic stages and supposed strict host specificity. The quantity of species with a limited number of overlapping diagnostic traits has led to a considerable mess in haemogregarine taxonomy and significant synonymy. We analysed host specificity, intra- and interspecific variability, evolutionary relationships, and the distribution of the type species of the genus Haemogregarina--H. stepanowi. The morphology of blood stages and 18S rDNA sequences of this haemogregarine from four western Palaearctic hard-shelled freshwater turtles (Emys orbicularis, Mauremys caspica, Mauremys leprosa and Mauremys rivulata) were compared with Haemogregarina balli. Additional sequences of 18S rDNA of Haemogregarina-like isolates collected from three species of African hinged terrapins (genus Pelusios) were used to enlarge the dataset for phylogenetic analyses. Thirteen sequences (1085 bp) of Haemogregarina representing all four western Palaearctic turtle species were identical, corresponding to H. stepanowi, which is closely related to the Nearctic species H. balli. In our analyses, Haemogregarina spp. constituted a monophyletic clade sister to the genus Hepatozoon. Haemogregarina stepanowi possesses a wide distribution range from the Maghreb, through Europe, Turkey and the Middle East to Iran. We consider that the genus Haemogregarina has a low host specificity crossing the family level of its vertebrate hosts and that its distribution is likely to be linked to the vector and definitive host--the leech.
- MeSH
- Eucoccidiida klasifikace cytologie genetika izolace a purifikace MeSH
- fylogeneze MeSH
- hostitelská specificita MeSH
- infekce přenášené vektorem MeSH
- interakce hostitele a parazita * MeSH
- kokcidióza parazitologie veterinární MeSH
- molekulární sekvence - údaje MeSH
- pijavice parazitologie MeSH
- ribozomální DNA chemie genetika MeSH
- RNA ribozomální 18S genetika MeSH
- sekvence nukleotidů MeSH
- sekvenční analýza DNA veterinární MeSH
- sladká voda MeSH
- želvy parazitologie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH