BACKGROUND: Cryptosporidium spp. are globally distributed parasites that infect epithelial cells in the microvillus border of the gastrointestinal tract of all classes of vertebrates. Cryptosporidium chipmunk genotype I is a common parasite in North American tree squirrels. It was introduced into Europe with eastern gray squirrels and poses an infection risk to native European squirrel species, for which infection is fatal. In this study, the biology and genetic variability of different isolates of chipmunk genotype I were investigated. METHODS: The genetic diversity of Cryptosporidium chipmunk genotype I was analyzed by PCR/sequencing of the SSU rRNA, actin, HSP70, COWP, TRAP-C1 and gp60 genes. The biology of chipmunk genotype I, including oocyst size, localization of the life cycle stages and pathology, was examined by light and electron microscopy and histology. Infectivity to Eurasian red squirrels and eastern gray squirrels was verified experimentally. RESULTS: Phylogenic analyses at studied genes revealed that chipmunk genotype I is genetically distinct from other Cryptosporidium spp. No detectable infection occurred in chickens and guinea pigs experimentally inoculated with chipmunk genotype I, while in laboratory mice, ferrets, gerbils, Eurasian red squirrels and eastern gray squirrels, oocyst shedding began between 4 and 11 days post infection. While infection in mice, gerbils, ferrets and eastern gray squirrels was asymptomatic or had mild clinical signs, Eurasian red squirrels developed severe cryptosporidiosis that resulted in host death. The rapid onset of clinical signs characterized by severe diarrhea, apathy, loss of appetite and subsequent death of the individual may explain the sporadic occurrence of this Cryptosporidium in field studies and its concurrent spread in the population of native European squirrels. Oocysts obtained from a naturally infected human, the original inoculum, were 5.64 × 5.37 μm and did not differ in size from oocysts obtained from experimentally infected hosts. Cryptosporidium chipmunk genotype I infection was localized exclusively in the cecum and anterior part of the colon. CONCLUSIONS: Based on these differences in genetics, host specificity and pathogenicity, we propose the name Cryptosporidium mortiferum n. sp. for this parasite previously known as Cryptosporidium chipmunk genotype I.

• Klíčová slova
• Biology, Course of infection, Cryptosporidiosis, Genetic diversity, Mortality, Oocyst size, Phylogeny,
• MeSH
• Cryptosporidiidae * MeSH
• Cryptosporidium * MeSH
• feces parazitologie   MeSH
• fretky MeSH
• fylogeneze MeSH
• genotyp MeSH
• Gerbillinae MeSH
• kryptosporidióza * parazitologie   MeSH
• kur domácí MeSH
• lidé MeSH
• morčata MeSH
• myši MeSH
• oocysty MeSH
• Sciuridae parazitologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• morčata MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

After invading the host organism, a battle occurs between the parasitic protists and the host's immune system, the result of which determines not only whether and how well the host survives and recovers, but also the fate of the parasite itself. The exact weaponry of this battle depends, among others, on the parasite localisation. While some parasitic protists do not invade the host cell at all (extracellular parasites), others have developed successful intracellular lifestyles (intracellular parasites) or attack only the surface of the host cell (epicellular parasites). Epicellular and intracellular protist parasites have developed various mechanisms to hijack host cell functions to escape cellular defences and immune responses, and, finally, to gain access to host nutrients. They use various evasion tactics to secure the tight contact with the host cell and the direct nutrient supply. This review focuses on the adaptations and evasion strategies of parasitic protists on the example of two very successful parasites of medical significance, Cryptosporidium and Leishmania, while discussing different localisation (epicellular vs. intracellular) with respect to the host cell.

• Klíčová slova
• Cryptosporidium, Leishmania, adaptation to parasitism, epicellular, evasion strategies, extracellular, host defence, intracellular, parasitic protist, parasitophorous sac, parasitophorous vacuole, unicellular parasite,
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Apicomplexa are unicellular eukaryotes that parasitise a wide spectrum of invertebrates and vertebrates, including humans. In their hosts, they occupy a variety of niches, from extracellular cavities (intestine, coelom) to epicellular and intracellular locations, depending on the species and/or developmental stages. During their evolution, Apicomplexa thus developed an exceptionally wide range of unique features to reach these diversified parasitic niches and to survive there, at least long enough to ensure their own transmission or that of their progeny. This review summarises the current state of knowledge on the attachment/invasive and nutrient uptake strategies displayed by apicomplexan parasites, focusing on trophozoite stages of their so far poorly studied basal representatives, which mostly parasitise invertebrate hosts. We describe their most important morphofunctional features, and where applicable, discuss existing major similarities and/or differences in the corresponding mechanisms, incomparably better described at the molecular level in the more advanced Apicomplexa species, of medical and veterinary significance, which mainly occupy intracellular niches in vertebrate hosts.

• Klíčová slova
• apical complex, attachment, epimerite, feeder organelle, mucron, myzocytosis, nutrition, parasitophorous vacuole/sac, pores, trophozoite,
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

IntroductionThis paper reviews the current knowledge and understanding of Cryptosporidium spp. and Giardia spp. in humans, animals and the environment in 10 countries in the eastern part of Europe: Bosnia and Herzegovina, Croatia, Czech Republic, Estonia, Hungary, Latvia, Poland, Romania, Serbia and Slovenia. Methods: Published scientific papers and conference proceedings from the international and local literature, official national health service reports, national databases and doctoral theses in local languages were reviewed to provide an extensive overview on the epidemiology, diagnostics and research on these pathogens, as well as analyse knowledge gaps and areas for further research. Results:Cryptosporidium spp. and Giardia spp. were found to be common in eastern Europe, but the results from different countries are difficult to compare because of variations in reporting practices and detection methodologies used. Conclusion: Upgrading and making the diagnosis/detection procedures more uniform is recommended throughout the region. Public health authorities should actively work towards increasing reporting and standardising reporting practices as these prerequisites for the reported data to be valid and therefore necessary for appropriate control plans.

• Klíčová slova
• One Health, cryptosporidiosis, giardiasis, zoonosis,
• MeSH
• Cryptosporidium genetika   izolace a purifikace   MeSH
• feces parazitologie   MeSH
• Giardia genetika   izolace a purifikace   MeSH
• giardiáza epidemiologie   parazitologie   MeSH
• kryptosporidióza epidemiologie   parazitologie   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé MeSH
• nemoci přenášené potravou epidemiologie   parazitologie   MeSH
• prevalence MeSH
• veřejné zdravotnictví * MeSH
• životní prostředí MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Geografické názvy
• východní Evropa epidemiologie   MeSH

The morphological, biological, and molecular characteristics of Cryptosporidium avian genotype V are described, and the species name Cryptosporidium avium is proposed to reflect its specificity for birds under natural and experimental conditions. Oocysts of C. avium measured 5.30-6.90 μm (mean = 6.26 μm) × 4.30-5.50 μm (mean = 4.86 μm) with a length to width ratio of 1.29 (1.14-1.47). Oocysts of C. avium obtained from four naturally infected red-crowned parakeets (Cyanoramphus novaezealandiae) were infectious for 6-month-old budgerigars (Melopsittacus undulatus) and hens (Gallus gallus f. domestica). The prepatent periods in both susceptible bird species was 11 days postinfection (DPI). The infection intensity of C. avium in budgerigars and hens was low, with a maximum intensity of 5000 oocysts per gram of feces. Oocysts of C. avium were microscopically detected at only 12-16 DPI in hens and 12 DPI in budgerigars, while PCR analyses revealed the presence of specific DNA in fecal samples from 11 to 30 DPI (the conclusion of the experiment). Cryptosporidium avium was not infectious for 8-week-old SCID and BALB/c mice (Mus musculus). Naturally or experimentally infected birds showed no clinical signs of cryptosporidiosis, and no pathology was detected. Developmental stages of C. avium were detected in the ileum and cecum using scanning electron microscopy. Phylogenetic analyses based on small subunit rRNA, actin, and heat shock protein 70 gene sequences revealed that C. avium is genetically distinct from previously described Cryptosporidium species.

• Klíčová slova
• Cryptosporidium avian genotype V, Cryptosporidium avium, Molecular analyses, Morphology, Transmission studies,
• MeSH
• cékum parazitologie   MeSH
• Cryptosporidium klasifikace   genetika   izolace a purifikace   MeSH
• feces parazitologie   MeSH
• fylogeneze MeSH
• ileum parazitologie   MeSH
• kryptosporidióza parazitologie   MeSH
• kur domácí parazitologie   MeSH
• Melopsittacus parazitologie   MeSH
• myši inbrední BALB C MeSH
• myši SCID MeSH
• myši MeSH
• nemoci drůbeže parazitologie   MeSH
• oocysty MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

This study aimed to evaluate and document the excystation process of Cryptosporidium muris oocysts in various incubation media, and to monitor the behaviour of excysting and freshly excysted sporozoites. A test of oocyst viability, using fluorescent double staining with fluorescein diacetate and propidium iodide, was performed prior to each experimental assay. Light microscope observations confirmed that relatively often only three sporozoites were released; the fourth one either left the oocyst later together with a residual body or remained trapped within the oocyst wall. These results suggest that successful oocyst excystation is not limited by the viability of all four sporozoites. Darkening of oocysts to opaque and their specific movement (the so-called "oocyst dancing") preceded the final excystation and liberation of sporozoites, while the dormant oocysts appeared refractive. The process of excystation in C. muris is not gradual as generally described in cryptosporidia but very rapid in an eruptive manner. Experiments were performed using oocysts stored at 4 °C for various time periods, as well as oocysts freshly shed from host rodents (Mastomys coucha) of different ages. The most suitable medium supporting high excystation rate (76 %) and prolonged motility of sporozoites was RPMI 1640, enriched with 5 % bovine serum albumin (BSA). Our results emphasize that to reliably evaluate the success of in vitro excystation of cryptosporidia, not only the number of released sporozoites in a set time period should be taken into consideration but also their subsequent activity (motility), as it is expected to be essential for the invasion of host cells.

• Klíčová slova
• Cryptosporidium muris, Excystation rate, Motility, Oocyst, Sporozoite, Viability test,
• MeSH
• Cryptosporidium účinky léků   fyziologie   MeSH
• krysa rodu Rattus MeSH
• kultivační média farmakologie   MeSH
• mikrobiální viabilita účinky léků   MeSH
• oocysty fyziologie   MeSH
• propidium MeSH
• sporozoiti účinky léků   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kultivační média MeSH
• propidium MeSH

• MeSH
• antigeny protozoální chemie   genetika   metabolismus   MeSH
• Apicomplexa imunologie   fyziologie   MeSH
• apikoplasty imunologie   fyziologie   MeSH
• biologická adaptace MeSH
• biologická evoluce MeSH
• druhová specificita MeSH
• extracelulární prostor imunologie   metabolismus   parazitologie   MeSH
• interakce hostitele a parazita * MeSH
• lidé MeSH
• membránové proteiny chemie   genetika   metabolismus   MeSH
• protozoální infekce zvířat imunologie   metabolismus   parazitologie   MeSH
• protozoální infekce imunologie   metabolismus   parazitologie   MeSH
• protozoální proteiny chemie   genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• antigeny protozoální MeSH
• membránové proteiny MeSH
• protozoální proteiny MeSH

This study focused on the attachment strategy, cell structure and the host-parasite interactions of the protococcidian Eleutheroschizon duboscqi, parasitising the polychaete Scoloplos armiger. The attached trophozoites and gamonts of E. duboscqi were detected at different development stages. The parasite develops epicellularly, covered by a host cell-derived, two-membrane parasitophorous sac forming a caudal tipped appendage. Staining with Evans blue suggests that this tail is protein-rich, supported by the presence of a fibrous substance in this area. Despite the ultrastructural evidence for long filaments in the tail, it stained only weakly for F-actin, while spectrin seemed to accumulate in this area. The attachment apparatus consists of lobes arranged in one (trophozoites) or two (gamonts) circles, crowned by a ring of filamentous fascicles. During trophozoite maturation, the internal space between the parasitophorous sac and parasite turns translucent, the parasite trilaminar pellicle seems to reorganise and is covered by a dense fibrous glycocalyx. The parasite surface is organised in broad folds with grooves in between. Micropores are situated at the bottom of the grooves. A layer of filaments organised in bands, underlying the folds and ending above the attachment fascicles, was detected just beneath the pellicle. Confocal microscopy, along with the application of cytoskeletal drugs (jasplakinolide, cytochalasin D, oryzalin) confirmed the presence of actin and tubulin polymerised forms in both the parasitophorous sac and the parasite, while myosin labelling was restricted to the sac. Despite positive tubulin labelling, no microtubules were detected in mature stages. The attachment strategy of E. duboscqi shares features with that of cryptosporidia and gregarines, i.e. the parasite itself conspicuously resembles an epicellularly located gregarine, while the parasitophorous sac develops in a similar manner to that in cryptosporidia. This study provides a re-evaluation of epicellular development in other apicomplexans and directly compares their niche with that of E. duboscqi.

• MeSH
• aktiny ultrastruktura   MeSH
• Apicomplexa klasifikace   fyziologie   ultrastruktura   MeSH
• interakce hostitele a parazita MeSH
• Polychaeta parazitologie   MeSH
• protozoální proteiny ultrastruktura   MeSH
• trofozoiti fyziologie   MeSH
• tubulin ultrastruktura   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aktiny MeSH
• protozoální proteiny MeSH
• tubulin MeSH

BACKGROUND: Gregarines represent a very diverse group of early emerging apicomplexans, parasitising numerous invertebrates and urochordates, and are considered of little practical significance. Recently, they have gained more attention since some analyses showed that cryptosporidia are more closely related to the gregarines than to coccidia. METHODOLOGY/PRINCIPAL FINDINGS: Using a combined microscopic approach, this study points out the spectacular strategy of Gregarina cuneata for attachment to host tissue and nutrient acquisition while parasitising the intestine of yellow mealworm larvae, and reveals the unusual dynamics of cellular interactions between the host epithelium and parasite feeding stages. Trophozoites of G. cuneata develop epicellularly, attached to the luminal side of the host epithelial cell by an epimerite exhibiting a high degree of morphological variability. The presence of contractile elements in the apical region of feeding stages indicates that trophozoite detachment from host tissue is an active process self-regulated by the parasite. A detailed discussion is provided on the possibility of reversible retraction and protraction of the eugregarine apical end, facilitating eventual reattachment to another host cell in better physiological conditions. The gamonts, found in contact with host tissue via a modified protomerite top, indicate further adaptation of parasite for nutrient acquisition via epicellular parasitism while keeping their host healthy. The presence of eugregarines in mealworm larvae even seems to increase the host growth rate and to reduce the death rate despite often heavy parasitisation. CONCLUSIONS/SIGNIFICANCE: Improved knowledge about the formation of host-parasite interactions in deep-branching apicomplexans, including gregarines, would offer significant insights into the fascinating biology and evolutionary strategy of Apicomplexa. Gregarines exhibit an enormous diversity in cell architecture and dimensions, depending on their parasitic strategy and the surrounding environment. They seem to be a perfect example of a coevolution between a group of parasites and their hosts.

• MeSH
• aktiny metabolismus   MeSH
• Apicomplexa cytologie   fyziologie   ultrastruktura   MeSH
• epitel metabolismus   parazitologie   ultrastruktura   MeSH
• epitelové buňky metabolismus   parazitologie   ultrastruktura   MeSH
• fyziologická adaptace * MeSH
• interakce hostitele a patogenu MeSH
• myosiny metabolismus   MeSH
• symbióza * MeSH
• Tenebrio parazitologie   MeSH
• transport proteinů MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aktiny MeSH
• myosiny MeSH