Hemolivia species infecting Central American wood turtles (Rhinoclemmys pulcherrima manni) and problems with differential diagnosis within the genus Hemolivia.

Blood parasites of the genus Hemolivia Petit, Landau, Baccam and Lainson, 1990 (Adeleorina: Karyolysidae) are hemogregarines of ectothermic vertebrates, such as lizards, chelonians, and toads. Only five species of Hemolivia from vertebrate hosts and one from their tick vector have been described so far. In the present study, Central American wood turtles (Rhinoclemmys pulcherrima manni) originating from Southern Nicaragua were screened for the presence of hemogregarines. Ten out of 30 specimens (33.3%) were positive for Hemolivia using both approaches - microscopy and PCR-based analyses. Phylogenetic analyses based on the 18S rRNA gene revealed the presence of two haplotypes, both placed as sister taxa in the Hemolivia clade. Their phylogenetic position was supported by high bootstrap values and high posterior probabilities, suggesting that there are at least two new distinct haplotypes corresponding to two distinct species. However, the specimens of each haplotype were microscopically indistinguishable from each other based on the gamont morphology, therefore, only a single species could be described and named, as Hemolivia pulcherrima n. sp. We consider that the uniform morphology of the most common blood stages of species of the genus Hemolivia complicates their differential diagnosis. Sequence divergence and different host spectra, therefore, remain the only differentiating tools.

Title: Espèces d'Hemolivia infectant les tortues peintes d'Amérique centrale (Rhinoclemmys pulcherrima manni) et problèmes de diagnostic différentiel au sein du genre Hemolivia. Abstract: Les parasites sanguins du genre Hemolivia Petit, Landau, Baccam et Lainson, 1990 (Adeleorina : Karyolysidae) sont des hémogrégarines de vertébrés ectothermes, tels que les lézards, les tortues et les crapauds. Seules cinq espèces d'Hemolivia provenant d'hôtes vertébrés et une de leur tique vectrice ont été décrites jusqu'à présent. Dans cette étude, des tortues peintes d'Amérique centrale (Rhinoclemmys pulcherrima manni) originaires du sud du Nicaragua ont été examinées pour détecter la présence d'hémogrégarines. Dix tortues sur 30 (33,3 %) étaient positives pour Hemolivia en utilisant les deux approches de microscopie et d'analyse de PCR. Les analyses phylogénétiques basées sur le gène de l'ARNr 18S ont révélé la présence de deux haplotypes, tous deux placés comme taxons frères dans le clade Hemolivia. Leur position phylogénétique était étayée par des valeurs de bootstrap et des probabilités postérieures élevées, suggérant qu'il existe au moins deux nouveaux haplotypes distincts correspondant à deux espèces distinctes. Cependant, les spécimens de chaque haplotype étaient impossibles à distinguer les uns des autres au microscope sur la base de la morphologie des gamontes. Par conséquent, une seule espèce a pu être décrite et nommée, comme Hemolivia pulcherrima n. sp. Nous considérons que l'uniformité de la morphologie des stades sanguins les plus courants des espèces du genre Hemolivia complique leur diagnostic différentiel. Les divergences de séquences et les différents spectres d'hôtes restent donc les seuls outils de différenciation.

European ground squirrels Spermophilus citellus (Linnaeus) do not share identical Cryptosporidium spp. with North American ground squirrels.

Cryptosporidium Tyzzer, 1910 is one of the most common protistan parasites of vertebrates. The results of this study provide the first data on Cryptosporidium diversity in the European ground squirrel Spermophilus citellus (Linnaeus). A total of 128 faecal samples of European ground squirrels from 39 localities in the Czech Republic were analysed for the presence of Cryptosporidium spp. by microscopy and PCR/sequence analysis of small subunit ribosomal RNA (SSU) and the actin gene. While the microscopical examination did not reveal the presence of any Cryptosporidium oocysts, eight samples from six localities were PCR-positive. Phylogenetic analyses revealed the presence of five different Cryptosporidium spp. isolates. Four isolates, designated as Cryptosporidium sp. isolate Sc01-04, detected in wild populations and never recorded before, clustered closely to Cryptosporidium genotypes that have previously been found in North American ground squirrels' species. Cryptosporidium sciurinum Prediger, Jezková, Holubová, Sak, Konecný, Rost, McEvoy, Rajský et Kvác, 2021 was found in an animal sanctuary. Because C. sciurinum had previously been detected in Eurasian red squirrels Sciurus vulgaris Linnaeus at the same facility, it can be concluded that this Cryptosporidium was transmitted from tree squirrels to ground squirrels within the animal sanctuary. The results indicate that populations of European and North American ground squirrels are parasitised by different Cryptosporidium spp. At the same time, this is the first description of the occurrence of C. sciurinum in ground squirrels.

Orthohantaviruses in Reservoir and Atypical Hosts in the Czech Republic: Spillover Infection and Indication of Virus-Specific Tissue Tropism.

Orthohantaviruses (genus Orthohantavirus) are a diverse group of viruses that are closely associated with their natural hosts (rodents, shrews, and moles). Several orthohantaviruses cause severe disease in humans. Central and western Europe are areas with emerging orthohantavirus occurrences. In our study, several orthohantaviruses, including the pathogenic Kurkino virus (KURV), were detected in their natural hosts trapped at several study sites in the Czech Republic. KURV was detected mainly in its typical host, the striped field mouse (Apodemus agrarius). Nevertheless, spillover infections were also detected in wood mice (Apodemus sylvaticus) and common voles (Microtus arvalis). Similarly, Tula virus (TULV) was found primarily in common voles, and events of spillover to rodents of other host species, including Apodemus spp., were recorded. In addition, unlike most previous studies, different tissues were sampled and compared to assess their suitability for orthohantavirus screening and possible tissue tropism. Our data suggest possible virus-specific tissue tropism in rodent hosts. TULV was most commonly detected in the lung tissue, whereas KURV was more common in the liver, spleen, and brain. Moreover, Seewis and Asikkala viruses were detected in randomly found common shrews (Sorex araneus). In conclusion, we have demonstrated the presence of human-pathogenic KURV and the potentially pathogenic TULV in their typical hosts as well as their spillover to atypical host species belonging to another family. Furthermore, we suggest the possibility of virus-specific tissue tropism of orthohantaviruses in their natural hosts. IMPORTANCE Orthohantaviruses (genus Orthohantavirus, family Hantaviridae) are a diverse group of globally distributed viruses that are closely associated with their natural hosts. Some orthohantaviruses are capable of infecting humans and causing severe disease. Orthohantaviruses are considered emerging pathogens due to their ever-increasing diversity and increasing numbers of disease cases. We report the detection of four different orthohantaviruses in rodents and shrews in the Czech Republic. Most viruses were found in their typical hosts, Kurkino virus (KURV) in striped field mice (Apodemus agrarius), Tula virus (TULV) in common voles (Microtus arvalis), and Seewis virus in common shrews (Sorex araneus). Nevertheless, spillover infections of atypical host species were also recorded for KURV, TULV, and another shrew-borne orthohantavirus, Asikkala virus. In addition, indications of virus-specific patterns of tissue tropism were observed. Our results highlight the circulation of several orthohantaviruses, including KURV, which is pathogenic to humans, among rodents and shrews in the Czech Republic.

Switch, disperse, repeat: host specificity is highly flexible in rodent-associated Eimeria.

Interplay between conserved host specificity and occasional host switches is an important process determining the evolution of host-parasite systems. Here, we address the dynamics of host switches at the population level in rodent-associated Eimeria. Focusing mainly on two ecologically similar host groups, Murinae and Arvicolinae, we show that the Eimeria infecting those hosts form a complex system of many genetic lineages with different host specificities. The broad geographic distribution of lineages indicates that they are well-established genetic forms which retained their host specificities while spreading across large geographic areas. We also demonstrate that genetic structure is only partially reflected by morphological traits.

Against growing synonymy: Identification pitfalls of Hepatozoon and Schellackia demonstrated on North Iranian reptiles.

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.

Eimeria sciurorum (Apicomplexa, Coccidia) From the Calabrian Black Squirrel (Sciurus meridionalis): An Example of Lower Host Specificity of Eimerians.

Host specificity plays one of the key roles in parasitism. It affects the evolution and diversification of both host and parasite, as well as it influences their geographical distribution, and epidemiological significance. For most of parasites, however, host specificity is unknown or misrepresented because it is difficult to be determined accurately. Here we provide the information about the lower host specificity of Eimeria sciurorum infecting squirrels, and its new host record for the Calabrian black squirrel Sciurus meridionalis, a southern Italian endemic species.

Generalist Eimeria species in rodents: Multilocus analyses indicate inadequate resolution of established markers.

Intracellular parasites of the genus Eimeria are described as tissue/host-specific. Phylogenetic classification of rodent Eimeria suggested that some species have a broader host range than previously assumed. We explore whether Eimeria spp. infecting house mice are misclassified by the most widely used molecular markers due to a lack of resolution, or whether, instead, these parasite species are indeed infecting multiple host species.With the commonly used markers (18S/COI), we recovered monophyletic clades of E. falciformis and E. vermiformis from Mus that included E. apionodes identified in other rodent host species (Apodemus spp., Myodes glareolus, and Microtus arvalis). A lack of internal resolution in these clades could suggest the existence of a species complex with a wide host range infecting murid and cricetid rodents. We question, however, the power of COI and 18S markers to provide adequate resolution for assessing host specificity. In addition to the rarely used marker ORF470 from the apicoplast genome, we present multilocus genotyping as an alternative approach. Phylogenetic analysis of 35 nuclear markers differentiated E. falciformis from house mice from isolates from Apodemus hosts. Isolates of E. vermiformis from Mus are still found in clusters interspersed with non-Mus isolates, even with this high-resolution data.In conclusion, we show that species-level resolution should not be assumed for COI and 18S markers in coccidia. Host-parasite cospeciation at shallow phylogenetic nodes, as well as contemporary coccidian host ranges more generally, is still open questions that need to be addressed using novel genetic markers with higher resolution.

Eimeria melogale n. sp. (Apicomplexa: Eimeriidae) in the Javan ferret-badger (Melogale orientalis).

The Javan ferret-badger Melogale orientalis (Carnivora: Mustelidae: Helictidinae) is a small carnivore endemic to Indonesia. In the family Mustelidae, 10 Eimeria, 12 Cystoisopora, one Isospora, and one Hammondia species are known, but no eimeriid coccidia has been yet described in the subfamily Helictinidae (ferret badgers). Coproscopic examination of Javan ferret-badgers imported into the Czech Republic revealed the presence of coccidian oocysts. Sporulated oocysts differ from other Eimeria known in the family Mustelidae by their small size (12.4-16.1 × 10.4-13.4 µm) and ovoidal shape. Morphological data and phylogenetic analyses of 18S rRNA and COI genes indicated a new species of Eimeria found in faecal samples of Javan ferret badgers. The species is described as E. melogale n. sp.

Gastrointestinal parasites of arctic foxes (Vulpes lagopus) and sibling voles (Microtus levis) in Spitsbergen, Svalbard.

The arctic fox (Vulpes lagopus), an apex predator with an omnipresent distribution in the Arctic, is a potential source of intestinal parasites that may endanger people and pet animals such as dogs, thus posing a health risk. Non-invasive methods, such as coprology, are often the only option when studying wildlife parasitic fauna. However, the detection and identification of parasites are significantly enhanced when used in combination with methods of molecular biology. Using both approaches, we identified unicellular and multicellular parasites in faeces of arctic foxes and carcasses of sibling voles (Microtus levis) in Svalbard, where molecular methods are used for the first time. Six new species were detected in the arctic fox in Svalbard, Eucoleus aerophilus, Uncinaria stenocephala, Toxocara canis, Trichuris vulpis, Eimeria spp., and Enterocytozoon bieneusi, the latter never found in the arctic fox species before. In addition, only one parasite was found in the sibling vole in Svalbard, the Cryptosporidium alticolis, which has never been detected in Svalbard before.

Isospora svecica sp. n. (Apicomplexa: Eimeriidae), a new species of coccidium from the white-spotted bluethroat Luscinia svecica cyanecula (Aves: Passeriformes: Muscicapidae).

Using a combination of morphological and molecular data, we describe a new apicomplexan parasite, Isospora svecica sp. n., from the white-spotted bluethroat, Luscinia svecica cyanecula, from the Czech Republic. Oocysts were found in its intestinal tract. Sporulation was exogenous and took 1-3 days. The oocysts were slightly ellipsoidal, of average size 26.17 × 20.33 µm, with a smooth bilayered wall. Micropyle, oocyst residuum, and polar granules were absent. Sporocysts were bottle-shaped, of an average size of 18.82 × 8.82 µm, with a thin, colourless wall. A conspicuous knob-like Stieda body was present. Substieda body was barely visible. Sporocyst residuum was present in the form of granules of various sizes. Sporozoites were banana-shaped and contained large anterior and small posterior refractile bodies. Partial DNA sequences of three genes were obtained from oocysts of Isospora svecica sp. n., being most closely related to other isosporans described from passerines. Little is known about the parasites of the avian family Muscicapidae, including coccidia, a highly prevalent parasitic protist group in all vertebrate classes. Only six species of the genus Isospora have so far been described in Muscicapidae, together with several "Isospora sp." that in fact most likely represent Isospora lacazei. The newly described Isospora svecica sp. n. differs morphologically from other coccidia reported from muscicapid birds, and represents the first coccidian species described from Luscinia svecica.

Detection and quantification of house mouse Eimeria at the species level - Challenges and solutions for the assessment of coccidia in wildlife.

Detection and quantification of coccidia in studies of wildlife can be challenging. Therefore, prevalence of coccidia is often not assessed at the parasite species level in non-livestock animals. Parasite species - specific prevalences are especially important when studying evolutionary questions in wild populations. We tested whether increased host population density increases prevalence of individual Eimeria species at the farm level, as predicted by epidemiological theory. We studied free-living commensal populations of the house mouse (Mus musculus) in Germany, and established a strategy to detect and quantify Eimeria infections. We show that a novel diagnostic primer targeting the apicoplast genome (Ap5) and coprological assessment after flotation provide complementary detection results increasing sensitivity. Genotyping PCRs confirm detection in a subset of samples and cross-validation of different PCR markers does not indicate bias towards a particular parasite species in genotyping. We were able to detect double infections and to determine the preferred niche of each parasite species along the distal-proximal axis of the intestine. Parasite genotyping from tissue samples provides additional indication for the absence of species bias in genotyping amplifications. Three Eimeria species were found infecting house mice at different prevalences: Eimeria ferrisi (16.7%; 95% CI 13.2-20.7), E. falciformis (4.2%; 95% CI 2.6-6.8) and E. vermiformis (1.9%; 95% CI 0.9-3.8). We also find that mice in dense populations are more likely to be infected with E. falciformis and E. ferrisi. We provide methods for the assessment of prevalences of coccidia at the species level in rodent systems. We show and discuss how such data can help to test hypotheses in ecology, evolution and epidemiology on a species level.

Isosporan Oocysts in the Faeces of Bank Voles (Myodes glareolus; Arvicolinae, Rodentia): Real Parasites, or Pseudoparasites?

Coccidia of the genus Isospora, their origin, taxonomy, and host specificity have been discussed for many years. The crucial point in question being the division of the genus, based on distinct evolutionary history and the presence/absence of the Stieda body, into the genera Isospora (Eimeriidae) parasitizing mainly birds and reptiles, and Cystoisospora (Sarcocystidae) parasitizing mammals. The description of the majority of Isospora species from rodents is based solely on the oocysts found in their faeces. Some of them have been described with the presence of the Stieda body, some without it, and, simultaneously, for all the described species the molecular data are entirely lacking. This study reveals the origin of isosporan oocysts found in faeces of bank voles based on morphological analyses, phylogenetic analyses, and experimental infections. Morphological analyses showed the presence of the Stieda body complex on sporocysts. Phylogenetic analyses demonstrated close phylogenetic relationships between Isospora from bank voles and avian isosporans. Experimental inoculations of bank voles with sporulated oocysts of Isospora did not result in the production of unsporulated oocysts. Hence, these organisms should be considered pseudoparasites of the bank voles/rodents (probably originating from avian Isospora species).

Intranuclear coccidiosis in tortoises - discovery of its causative agent and transmission.

Intranuclear coccidiosis of testudines (known as TINC) is an emerging disease in chelonians. Although endogenous stages were repeatedly detected in various tissues, attempts to find the oocysts in faeces failed, leaving the question of the transmission and classification of the causative agent of TINC unresolved. We recorded small spherical oocysts (∼6-7 µm in diameter) of an eimeriid coccidium in faeces of a leopard tortoise (Stigmochelys pardalis). Sporulated oocysts were used for the experimental oral inoculation of juvenile coccidia-free tortoises representing 5 species (S. pardalis, Testudo graeca, T. hermanni, T. horsfieldii, and Geochelone sulcata). The oocysts' association with TINC was confirmed based on clinical signs, histopathological findings of intranuclear endogenous stages of the coccidium in many organs (including intestine), and by the partial 18S rDNA sequence analysis of the DNA isolated from organs of the experimentally infected animals and from a single naturally infected as well as from all experimentally infected tortoises. Breeding colonies of chelonians should be screened for this pathogen in order to prevent its further spread and unwanted introduction into endangered free-ranging chelonian populations.

Two new species of Eimeria (Apicomplexa: Eimeriidae) in Philippine tarsier (Tarsius syrichta).

Philippine tarsier (Tarsius syrichta) is a small nocturnal primate from the Philippines. Little is known about tarsier parasites, including coccidia (Apicomplexa: Eimeriidae), a highly prevalent parasitic protist group in all vertebrate classes. Only 7 valid species of the genus Eimeria, seven species of Isospora and 5 species of Cyclospora have been described in Primates. This study extends the number of coccidia known in primates by two new species obtained from faeces of Philippine tarsiers from Bohol Island. The newly described Eimeria syrichta n. sp. and Eimeria boholensis n. sp. differ morphologically from each other as well as from other coccidia reported from primates. Partial DNA sequences of three genes were obtained from oocysts of E. syrichta n. sp. and E. boholensis n. sp., and formed clusters according to their host specificity; however, there are no other sequentional data of coccidia from primates, except for the genus Cyclospora, which clusters inside the chicken eimerians, and Cystoisospora belli, which is phylogenetically related to Sarcocystidae. More molecular data on coccidia infecting primates are needed for further discussion.

Mysteries of host switching: Diversification and host specificity in rodent-coccidia associations.

Recent studies show that host switching is much more frequent than originally believed and constitutes an important driver in evolution of host-parasite associations. However, its frequency and ecological mechanisms at the population level have been rarely investigated. We address this issue by analyzing phylogeny and population genetics of an extensive sample, from a broad geographic area, for commonly occurring parasites of the genus Eimeria within the abundant rodent genera Apodemus, Microtus and Myodes, using two molecular markers. At the most basal level, we demonstrate polyphyletic arrangement, i.e. multiple origin, of the rodent-specific clusters within the Eimeria phylogeny, and strong genetic/phylogenetic structure within these lineages determined at least partially by specificities to different host groups. However, a novel and the most important observation is a repeated occurrence of host switches among closely related genetic lineages which may become rapidly fixed. Within the studied model, this phenomenon applies particularly to the switches between the eimerians from Apodemus flavicollis/Apodemus sylvaticus and Apodemus agrarius groups. We show that genetic differentiation and isolation between A. flavicollis/A. sylvaticus and A. agrarius faunas is a secondary recent event and does not reflect host-parasite coevolutionary history. Rather, it provides an example of rapid ecology-based differentiation in the parasite population.

Phylogeny, Diversity, Distribution, and Host Specificity of Haemoproteus spp. (Apicomplexa: Haemosporida: Haemoproteidae) of Palaearctic Tortoises.

A complex wide-range study on the haemoproteid parasites of chelonians was carried out for the first time. Altogether, 811 samples from four tortoise species from an extensive area between western Morocco and eastern Afghanistan and between Romania and southern Syria were studied by a combination of microscopic and molecular-genetic methods. Altogether 160 Haemoproteus-positive samples were gathered in the area between central Anatolia and eastern Afghanistan. According to variability in the cytochrome b gene, two monophyletic evolutionary lineages were distinguished; by means of microscopic analysis it was revealed that they corresponded to two previously described species-Haemoproteus anatolicum and Haemoproteus caucasica. Their distribution areas overlap only in a narrow strip along the Zagros Mts. range in Iran. This fact suggests the involvement of two different vector species with separated distribution. Nevertheless, no vectors were confirmed. According to phylogenetic analyses, H. caucasica represented a sister group to H. anatolicum, and both of them were most closely related to H. pacayae and H. peltocephali, described from South American river turtles. Four unique haplotypes were revealed in the population of H. caucasica, compared with seven haplotypes in H. anatolicum. Furthermore, H. caucasica was detected in two tortoise species, Testudo graeca and Testudo horsfieldii, providing evidence that Haemoproteus is not strictly host-specific to the tortoise host species.

Haemogregarines of freshwater turtles from Southeast Asia with a description of Haemogregarina sacaliae sp. n. and a redescription of Haemogregarina pellegrini Laveran and Pettit, 1910.

The uniform morphology of the developmental stages of Haemogregarina species and the insufficient information supplied by the simplistic descriptions of previous authors complicates their differential diagnosis and proper species identification. In this study, we detected Haemogregarina spp. in 6 out of 22 (27.2%) examined turtles originating from Southeast Asia, Malayemys subtrijuga (n = 4), Sacalia quadriocellata (n = 1) and Platysternon megacephalum (n = 1), and compared them with the available literature data. Microscopic analysis of our isolates distinguished 2 morphological species, Haemogregarina pellegrini and one new species, being described in this paper as Haemogregarina sacaliae sp. n. Phylogenetic analyses based on 1210 bp long fragment of 18S rDNA sequences placed both haemogregarines firmly within the monophyletic Haemogregarina clade. Isolates of H. pellegrini from 2 distantly related turtle hosts, M. subtrijuga and P. megacephalum, were genetically identical. Despite the fact that numerous Haemogregarina species of turtles have been described, the incompleteness of the morphological data and relatively low host specificity provides the space for large synonymy within this taxon. Therefore, a complex approach combining microscopic analyses together with molecular-genetic methods should represent the basic standard for all taxonomic studies.

Hemolivia and hepatozoon: haemogregarines with tangled evolutionary relationships.

The generic name Hemolivia has been used for haemogregarines characterized by morphological and biological features. The few molecular studies, focused on other haemogregarine genera but involving Hemolivia samples, indicated its close relationship to the genus Hepatozoon. Here we analyze molecular data for Hemolivia from a broad geographic area and host spectrum and provide detailed morphological documentation of the included samples. Based on molecular analyses in context of other haemogregarines, we demonstrate that several sequences deposited in GenBank from isolates described as Hepatozoon belong to the Hemolivia cluster. This illustrates the overall difficulty with recognizing Hemolivia and Hepatozoon without sufficient morphological and molecular information. The close proximity of both genera is also reflected in uncertainty about their precise phylogeny when using 18S rDNA. They cluster with almost identical likelihood either as two sister taxa or as monophyletic Hemolivia within paraphyletic Hepatozoon. However, regardless of these difficulties, the results presented here provide a reliable background for the unequivocal placement of new samples into the Hemolivia/ Hepatozoon complex.

Haemogregarines from western Palaearctic freshwater turtles (genera Emys, Mauremys) are conspecific with Haemogregarina stepanowi Danilewsky, 1885.

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.

Evolutionary plasticity in coccidia - striking morphological similarity of unrelated coccidia (apicomplexa) from related hosts: Eimeria spp. from African and Asian Pangolins (Mammalia: Pholidota).

Two morphologically similar, but phylogenetically unrelated Eimeria species from ancient mammals, African Tree Pangolin Phataginus tricuspis and Sunda Pangolin Manis javanica (Pholidota: Manidae), from two distant biogeographic realms (Afrotropical and Oriental), are characterized and compared morphologically and molecularly. Phylogenetic analyses produced an unstable topology. However, while precise position of the two Eimeria species from pangolins could not be firmly established due to the lack of related taxa, it is evident that they are not closely related and do not fall into any of the so far recognized eimerian lineages. Moreover, an eimerian found in P. tricuspis is described as a new species Eimeria nkaka n. sp., based on morphology of oocysts, endogenous developmental stages and sequence data.