OBJECTIVES: Studies on the pathogenesis and immune responses of Cryptosporidium infection and development of drugs and vaccines use mostly immunocompromised mouse models. In this study, we establish an immunocompetent mouse model of cryptosporidiosis with high intensity and long duration of infection. METHODS: We have obtained a Cryptosporidium tyzzeri isolate from laboratory mice, and infect adult C57BL/6 J mice experimentally with the isolate for determinations of infectivity, infection patterns, pathological changes, and transcriptomic responses. RESULTS: The isolate has an ID50 of 5.2 oocysts, with oocyst shedding lasting at high levels for >2 months. The oocyst shedding is boosted by immunosuppression of animals and suppressed by paromomycin treatment. The isolate induces strong inflammatory and acquired immune responses, but down-regulates the expression of α-defensins in epithelium. Comparative genomics analysis has revealed significant sequence differences from other isolates in subtelomeric genes. The down-regulation of the expression of α-defensins may be responsible for the high-intensity and long-lasting infection in this animal model. CONCLUSIONS: The immunocompetent mouse model of cryptosporidiosis developed has the advantages of high oocyst shedding intensity and long oocyst shedding duration. It provides an effective mechanism for the propagation of Cryptosporidium, evaluations of potential therapeutics, and studies of pathogen biology and immune responses.

• Klíčová slova
• Animal model, C57BL/6J mice, Cryptosporidiosis, Cryptosporidium, Immune responses, α-defensins,
• MeSH
• alfa-defensiny * MeSH
• Cryptosporidium parvum * MeSH
• Cryptosporidium * fyziologie   MeSH
• feces MeSH
• kryptosporidióza * patologie   MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• oocysty MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• alfa-defensiny * MeSH

Long-tailed chinchillas Chinchilla lanigera are popular rodent species kept both in households, where they are hand-raised as pets, and in zoological facilities. From January 2016 to February 2017, 13 juvenile chinchillas from five facilities in Japan were diagnosed with cryptosporidiosis at the animal hospital. Eight of the cases were fatal. All of the animals were imported from the Czech Republic by the same vendor. Histopathological and multilocus sequence analyses using 18S ribosomal RNA, actin, 70-kDa heat shock protein, and 60-kDa glycoprotein genes confirmed Cryptosporidium ubiquitum of subtype XIId as the etiological agent. Multilocus analysis demonstrated the presence of two new sequence types closely related to the C. ubiquitum Xlld strain isolated from a human in the USA. This study indicated that potentially zoonotic Cryptosporidium is widespread and may have caused a high number of deaths among imported juvenile chinchillas.

• Klíčová slova
• Cryptosporidiosis, Emerging infectious disease, Multilocus sequence analysis, Subtype XIId, Zoonotic infection,
• MeSH
• činčila parazitologie   MeSH
• Cryptosporidium genetika   izolace a purifikace   MeSH
• feces parazitologie   MeSH
• fylogeneze MeSH
• genotyp MeSH
• hospodářská zvířata parazitologie   MeSH
• importované infekce epidemiologie   mortalita   parazitologie   MeSH
• kryptosporidióza epidemiologie   mortalita   patologie   přenos   MeSH
• multilokusová sekvenční typizace MeSH
• objevující se infekční nemoci epidemiologie   parazitologie   MeSH
• protozoální DNA genetika   MeSH
• RNA ribozomální 18S genetika   MeSH
• zoonózy epidemiologie   parazitologie   přenos   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Japonsko epidemiologie   MeSH
• Názvy látek
• protozoální DNA MeSH
• RNA ribozomální 18S MeSH

Cryptosporidium parvum VF383 has been reported in humans, domesticated ruminants, and wild rats worldwide and described under several names including Cryptosporidium suis-like, based on its close phylogenetic relationship to C. suis. Unlike C. suis, however, it has never been detected in pigs. In the present work, C. parvum VF383 originating from wild brown rats was not infectious for piglets or calves but was infectious for laboratory brown rats, BALB/c mice, and Mongolian gerbils. The prepatent period was 4-5 days for all rodents. The patent period was longer for rats (>30 days) than other rodents (<20 days). None of the rodents developed clinical signs of infection. In all rodents, life cycle stages were detected in the colon by histology and electron microscopy. Oocysts were morphometrically similar to those of C. parvum and smaller than those of C. suis, measuring 5.20 × 4.94 μm. Phylogenetic analyses of 18S rRNA, actin, and HSP70 gene sequences revealed C. parvum VF383 to be genetically distinct from, C. suis, and other described species of Cryptosporidium. Morphological, genetic, and biological data support the establishment of C. parvum VF383 as a new species, and we propose the name Cryptosporidium occultus sp. n.

• Klíčová slova
• Histology, Molecular phylogeny, Morphometry, New species, Transmission studies,
• MeSH
• aktiny genetika   MeSH
• Cryptosporidium klasifikace   genetika   MeSH
• druhová specificita MeSH
• kolon parazitologie   MeSH
• kryptosporidióza parazitologie   patologie   MeSH
• krysa rodu Rattus MeSH
• proteiny tepelného šoku HSP70 genetika   MeSH
• RNA ribozomální 18S genetika   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aktiny MeSH
• proteiny tepelného šoku HSP70 MeSH
• RNA ribozomální 18S MeSH

Faecal samples from striped field mice (n = 72) and yellow-necked mice (n = 246) were screened for Cryptosporidium by microscopy and PCR/sequencing. Phylogenetic analysis of small-subunit rRNA, Cryptosporidium oocyst wall protein and actin gene sequences revealed the presence of C. parvum, C. hominis, C. muris and two new species, C. apodemi and C. ditrichi. Oocysts of C. apodemi are smaller than C. ditrichi and both are experimentally infectious for yellow-necked mice but not for common voles. Additionally, infection by C. ditrichi was established in one of three BALB/c mice. The prepatent period was 7-9 and 5-6 days post infection for C. apodemi and C. ditrichi, respectively. The patent period was greater than 30 days for both species. Infection intensity of C. ditrichi ranged from 4000-50,000 oocyst per gram of faeces and developmental stages of C. ditrichi were detected in the jejunum and ileum. In contrast, neither oocysts nor endogenous developmental stages of C. apodemi were detected in faecal or tissue samples, although C. apodemi DNA was detected in contents from the small and large intestine. Morphological, genetic, and biological data support the establishment of C. apodemi and C. ditrichi as a separate species of the genus Cryptosporidium.

• Klíčová slova
• Europe, Experimental infection, Molecular analyses, Oocyst size, Phylogeny, Rodentia,
• MeSH
• aktiny genetika   MeSH
• Cryptosporidium klasifikace   genetika   izolace a purifikace   MeSH
• druhová specificita MeSH
• feces parazitologie   MeSH
• fylogeneze * MeSH
• kryptosporidióza parazitologie   patologie   MeSH
• Murinae parazitologie   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• protozoální proteiny genetika   MeSH
• ribozomální DNA genetika   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aktiny MeSH
• oocyst wall protein, Cryptosporidium MeSH Prohlížeč
• protozoální proteiny MeSH
• ribozomální DNA MeSH

This study describes cryptosporidiosis in an overwintering group of 15 European hedgehogs (Erinaceus europaeus), comprising 3 adults and 12 juveniles. Four juvenile hedgehogs were hospitalised with anorexia, malodorous diarrhoea and dehydration. Immediate parasitological examinations revealed the presence of Cryptosporidium sp. in these animals and also in 5 other juveniles. All hedgehogs were coproscopically monitored for 4 months over the winter season. Shedding of Cryptosporidium oocysts persisted from 6 to 70 days. Repeated shedding of Cryptosporidium oocysts occurred in 3 animals after 4 months subsequent to the first outbreak. Clinical signs were observed only at the beginning of the outbreak (apathy, anorexia, general weakness, mild dehydration, and malodorous faeces with changed consistence - soft/diarrhoea) in the 4 hospitalised juveniles. Overall 11 hedgehogs were Cryptosporidium-positive, both microscopically and by PCR methods. Sequence analyses of SSU rRNA and gp60 genes revealed the presence of C. parvum IIdA18G1 subtype in all positive hedgehogs. Moreover, 3 hedgehogs had a mixed infection of the zoonotic C. parvum and C. erinacei XIIIaA19R13 subtype. Cryptosporidium infections can be rapidly spread among debilitated animals and the positive hedgehogs released back into the wild can be a source of the infection for individuals weakened after hibernation.

• Klíčová slova
• Cryptosporidiosis, Erinaceus europaeus, European hedgehog,
• MeSH
• Cryptosporidium parvum klasifikace   genetika   fyziologie   MeSH
• Cryptosporidium klasifikace   genetika   fyziologie   MeSH
• feces parazitologie   MeSH
• fylogeneze MeSH
• geny rRNA genetika   MeSH
• ježkovití parazitologie   MeSH
• kryptosporidióza parazitologie   patologie   MeSH
• nemocnice veterinární MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH

Piglets from 4 to 8 weeks of age originated from a Cryptosporidium-free research breed were orally inoculated with 1 × 10(6) infectious oocysts of Cryptosporidium scrofarum. The number of shed oocysts per gram of faeces served to describe the infection intensity and prepatent period. In addition, faecal samples collected daily and tissue samples of the small and large intestine collected at 30 days post-inoculation were examined for the C. scrofarum small subunit ribosomal RNA gene using PCR. The piglets inoculated at 4-weeks of age remained uninfected, whereas 5-week-old and older animals were fully susceptible with a prepatent period ranging from 4 to 8 days. Susceptible pigs shed oocysts intermittently, and shedding intensity, reaching a mean maximum of 6000 oocysts per gram, did not differ significantly among infected animals. This study demonstrates that pigs become susceptible to C. scrofarum infection as late as 5-weeks of age. The mechanisms of age related susceptibility remain unknown.

• Klíčová slova
• Cryptosporidium scrofarum, Infection, Molecular analyses, Pigs, Susceptibility, Transmission studies,
• MeSH
• Cryptosporidium genetika   MeSH
• feces parazitologie   MeSH
• geny rRNA genetika   MeSH
• kryptosporidióza imunologie   patologie   veterinární   MeSH
• nemoci prasat imunologie   patologie   MeSH
• počet parazitárních vajíček MeSH
• prasata MeSH
• věkové faktory MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

This study focuses on mapping the life cycle of Cryptosporidium muris in two laboratory rodents; BALB/c mice and the southern multimammate rat Mastomys coucha, differing in their prepatent and patent periods. Both rodents were simultaneously experimentally inoculated with viable oocysts of C. muris (strain TS03). Animals were dissected and screened for the presence of the parasite using a combined morphological approach and nested PCR (SSU rRNA) at different times after inoculation. The occurrence of first developmental stages of C. muris in stomach was detected at 2.5 days post-infection (dpi). The presence of Type II merogony, appearing 36 h later than Type I merogony, was confirmed in both rodents. Oocysts exhibiting different size and thickness of their wall were observed from 5 dpi onwards in stomachs of both host models. The early phase of parasitization in BALB/c mice progressed rapidly, with a prepatent period of 7.5-10 days; whereas in M. coucha, the developmental stages of C. muris were first observed 12 h later in comparison with BALB/c mice and prepatent period was longer (18-21 days). Similarly, the patent periods of BALB/c mice and M. coucha differed considerably, i.e. 10-15 days vs chronic infection throughout the life of the host, respectively.

• MeSH
• Cryptosporidium růst a vývoj   fyziologie   MeSH
• druhová specificita MeSH
• feces parazitologie   MeSH
• kryptosporidióza parazitologie   patologie   MeSH
• modely nemocí na zvířatech MeSH
• Murinae MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• oocysty MeSH
• stadia vývoje * MeSH
• trofozoiti MeSH
• žaludeční sliznice patologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

We describe the morphological, biological, and molecular characteristics of Cryptosporidium pig genotype II and propose the species name Cryptosporidium scrofarum n. sp. to reflect its prevalence in adult pigs worldwide. Oocysts of C. scrofarum are morphologically indistinguishable from C. parvum, measuring 4.81-5.96 μm (mean=5.16)×4.23-5.29 μm (mean=4.83) with a length to width ratio of 1.07±0.06 (n=400). Oocysts of C. scrofarum obtained from a naturally infected pig were infectious for 8-week-old pigs but not 4-week-old pigs. The prepatent period in 8-week-old Cryptosporidium-naive pigs was 4-6 days and the patent period was longer than 30 days. The infection intensity of C. scrofarum in pigs was generally low, in the range 250-4000 oocysts per gram of feces. Infected pigs showed no clinical signs of cryptosporidiosis and no pathology was detected. Cryptosporidium scrofarum was not infectious for adult SCID mice, adult BALB/c mice, Mongolian gerbils (Meriones unguiculatus), southern multimammate mice (Mastomys coucha), yellow-necked mice (Apodemus flavicollis), or guinea pigs (Cavia porcellus). Phylogenetic analyses based on small subunit rRNA, actin, and heat shock protein 70 gene sequences revealed that C. scrofarum is genetically distinct from all known Cryptosporidium species.

• MeSH
• Cryptosporidium klasifikace   cytologie   genetika   MeSH
• druhová specificita MeSH
• feces parazitologie   MeSH
• fylogeneze * MeSH
• genotyp MeSH
• Gerbillinae MeSH
• kryptosporidióza patologie   veterinární   MeSH
• morčata MeSH
• myši inbrední BALB C MeSH
• myši SCID MeSH
• myši MeSH
• nemoci prasat parazitologie   patologie   MeSH
• prasata MeSH
• protozoální geny genetika   MeSH
• zvířata MeSH
• Check Tag
• morčata MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

We report a case of severe human cryptosporidiosis caused by Cryptosporidium tyzzeri and C. parvum with an unusually high frequency of liquid stools. Wild mice were the most likely source of infection, demonstrating the potential for wild-mouse-borne Cryptosporidium to infect humans and highlighting the health risks associated with synantropic rodents.

• MeSH
• Cryptosporidium klasifikace   genetika   izolace a purifikace   MeSH
• divoká zvířata MeSH
• dospělí MeSH
• kryptosporidióza diagnóza   parazitologie   patologie   přenos   MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• myši MeSH
• protozoální DNA chemie   genetika   MeSH
• sekvenční analýza DNA MeSH
• zoonózy parazitologie   přenos   MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• protozoální DNA MeSH

The infectivity of Cryptosporidium muris and Cryptosporidium andersoni in various species of voles was studied using experimental infections. None of the experimental voles inoculated with 1 × 10(5) oocysts of Cryptosporidium spp. shed any oocysts during 40 DPI, except Brandt's vole (Lasiopodomys brandtii), which was susceptible to C. muris infection. Experiments confirmed the resistance of voles of the genus Microtus sensu stricto to infection with mammalian gastric cryptosporidia, which provides a new study model with prospects to more fully understand the processes involved in the phenomenon of host specificity of this group of protists.

• MeSH
• Arvicolinae parazitologie   MeSH
• Cryptosporidium patogenita   fyziologie   MeSH
• hostitelská specificita MeSH
• kryptosporidióza parazitologie   patologie   MeSH
• modely nemocí na zvířatech MeSH
• odolnost vůči nemocem MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH