Cryptosporidium spp., Giardia intestinalis and microsporidia are unicellular opportunistic pathogens that can cause gastrointestinal infections in both animals and humans. Since companion animals may serve as a source of infection, the aim of the present screening study was to analyse the prevalence of these intestinal protists in fecal samples collected from dogs living in 10 animal shelters in central Europe (101 dogs from Poland and 86 from the Czech Republic), combined with molecular subtyping of the detected organisms in order to assess their genetic diversity. Genus-specific polymerase chain reactions were performed to detect DNA of the tested species and to conduct molecular subtyping in collected samples, followed by statistical evaluation of the data obtained (using χ2 or Fisher's tests). The observed prevalence was 15.5, 10.2, 1 and 1% for G. intestinalis, Enterocytozoon bieneusi, Cryptosporidium spp. and Encephalitozoon cuniculi, respectively. Molecular evaluation has revealed the predominance of dog-specific genotypes (Cryptosporidium canis XXe1 subtype; G. intestinalis assemblages C and D; E. cuniculi genotype II; E. bieneusi genotypes D and PtEbIX), suggesting that shelter dogs do not pose a high risk of human transmission. Interestingly, the percentage distribution of the detected pathogens differed between both countries and individual shelters, suggesting that the risk of infection may be associated with conditions typical of a given location.

• Klíčová slova
• 60-kDa glycoprotein, PCR, glutamate dehydrogenase, internal transcribed spacer region of rRNA, intestinal protists, opportunistic pathogens, small ribosomal subunit rRNA, subtyping, triosephosphate isomerase, β-giardin,
• MeSH
• Cryptosporidium * genetika   izolace a purifikace   klasifikace   MeSH
• Enterocytozoon * genetika   izolace a purifikace   klasifikace   MeSH
• feces * parazitologie   mikrobiologie   MeSH
• genotyp MeSH
• Giardia lamblia genetika   izolace a purifikace   klasifikace   MeSH
• Giardia genetika   izolace a purifikace   klasifikace   MeSH
• giardiáza * veterinární   epidemiologie   parazitologie   MeSH
• hostitelská specificita MeSH
• kryptosporidióza * epidemiologie   parazitologie   MeSH
• mikrosporidióza * veterinární   epidemiologie   MeSH
• nemoci psů * parazitologie   epidemiologie   mikrobiologie   MeSH
• prevalence MeSH
• psi MeSH
• zvířata MeSH
• Check Tag
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Polsko epidemiologie   MeSH

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

Cryptosporidium spp. are common protozoan pathogens in mammals. With pet rodents being integrated into modern life, the potential roles of them in transmitting parasites to humans need assessments. In the present study, we examined the occurrence of Cryptosporidium spp. in pet rodents in Guangdong, south China. A total of 697 fecal samples were collected from 11 species of rodents in seven pet shops, one pet market and one farm. Cryptosporidium spp. were identified by PCR analysis of the small subunit rRNA gene. An overall infection rate of 36.9% (257/697) was obtained, with infection rates varying from 9.3% in chinchillas, 52.3% in guinea pigs, 57.1% in squirrels, to 68.4% in cricetid animals. Nine Cryptosporidium species and genotypes were identified, including C. wrairi (in 129 guinea pigs), C. andersoni (in 34 hamsters), C. homai (in 32 guinea pigs), Cryptosporidium hamster genotype (in 30 hamsters), C. ubiquitum (in 24 chinchillas and squirrels), C. parvum (in 2 chinchillas), Cryptosporidium ferret genotype (in 2 chipmunks), C. muris (in 1 hamster and 1 guinea pig), and Cryptosporidium chipmunk genotype V (in 1 chinchilla and 1 chipmunk). Sequence analysis of the 60 kDa glycoprotein gene identified three subtype families of C. ubiquitum, including family XIId in 15 chinchillas, XIIa in 5 chinchillas, and a new subtype family (XIIi) in 1 squirrel. The identification of C. parvum and C. ubiquitum in pet rodents suggests that these animals, especially chinchillas, could serve as reservoirs of human-pathogenic Cryptosporidium spp. Hygiene should be practiced in the rear and care of these animals, and One Health measures should be developed to reduce the occurrence of zoonotic Cryptosporidium infections due to contact with pet rodents.

• Klíčová slova
• Cryptosporidium, Molecular epidemiology, One health, Pet rodents, Zoonosis,
• Publikační typ
• časopisecké články MeSH

Cryptosporidium spp., common parasites of vertebrates, remain poorly studied in wildlife. This study describes the novel Cryptosporidium species adapted to nutrias (Myocastor coypus). A total of 150 faecal samples of feral nutria were collected from locations in the Czech Republic and Slovakia and examined for Cryptosporidium spp. oocysts and specific DNA at the SSU, actin, HSP70, and gp60 loci. Molecular analyses revealed the presence of C. parvum (n = 1), C. ubiquitum subtype family XIId (n = 5) and Cryptosporidium myocastoris n. sp. XXIIa (n = 2), and XXIIb (n = 3). Only nutrias positive for C. myocastoris shed microscopically detectable oocysts, which measured 4.8-5.2 × 4.7-5.0 µm, and oocysts were infectious for experimentally infected nutrias with a prepatent period of 5-6 days, although not for mice, gerbils, or chickens. The infection was localised in jejunum and ileum without observable macroscopic changes. The microvilli adjacent to attached stages responded by elongating. Clinical signs were not observed in naturally or experimentally infected nutrias. Phylogenetic analyses at SSU, actin, and HSP70 loci demonstrated that C. myocastoris n. sp. is distinct from other valid Cryptosporidium species.

• Klíčová slova
• adaptation, biology, course of infection, infectivity, oocyst size, parasite, phylogeny, prevalence,
• Publikační typ
• časopisecké články MeSH

The diversity and biology of Cryptosporidium that is specific for rats (Rattus spp.) are not well studied. We examined the occurrence and genetic diversity of Cryptosporidium spp. in wild brown rats (Rattus norvegicus) by microscopy and polymerase chain reaction (PCR)/sequencing targeting the small subunit rDNA (SSU), actin and HSP70 genes. Out of 343 faecal samples tested, none were positive by microscopy and 55 were positive by PCR. Sequence analysis of SSU gene revealed the presence of Cryptosporidium muris (n = 4), C. andersoni (n = 3), C. ryanae (n = 1), C. occultus (n = 3), Cryptosporidium rat genotype I (n = 23), Cryptosporidium rat genotype IV (n = 16) and novel Cryptosporidium rat genotype V (n = 5). Spherical oocysts of Cryptosporidium rat genotype I obtained from naturally-infected rats, measuring 4.4-5.4 μm × 4.3-5.1 μm, were infectious to the laboratory rats, but not to the BALB/c mice (Mus musculus) nor Mongolian gerbils (Meriones unguiculatus). The prepatent period was 3 days post infection and the patent period was longer than 30 days. Naturally- and experimentally-infected rats showed no clinical signs of disease. Percentage of nucleotide similarities at the SSU, actin, HSP70 loci between C. ratti n. sp. and the rat derived C. occultus and Cryptosporidium rat genotype II, III, IV, and V ranged from 91.0 to 98.1%. These genetic variations were similar or greater than that observed between closely related species, i.e. C. parvum and C. erinacei (93.2-99.5%). Our morphological, genetic and biological data support the establishment of Cryptosporidium rat genotype I as a new species, Cryptosporidium ratti n. sp.

• Klíčová slova
• Cryptosporidium ratti, infectivity, morphometric analysis, phylogeny, prevalence,
• MeSH
• aktiny genetika   MeSH
• Cryptosporidium * klasifikace   genetika   izolace a purifikace   MeSH
• divoká zvířata parazitologie   MeSH
• feces parazitologie   MeSH
• fylogeneze MeSH
• genetická variace MeSH
• klasifikace MeSH
• krysa rodu Rattus parazitologie   MeSH
• myši MeSH
• prevalence MeSH
• proteiny tepelného šoku HSP70 genetika   MeSH
• protozoální DNA MeSH
• ribozomální DNA genetika   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus parazitologie   MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aktiny MeSH
• proteiny tepelného šoku HSP70 MeSH
• protozoální DNA MeSH
• ribozomální DNA MeSH

BACKGROUND: Bamboo rats are widely farmed in southern China for meat, but their potential in transmitting pathogens to humans and other farm animals remains unclear. METHODS: To understand the transmission of Cryptosporidium spp. in these animals, 709 fecal samples were collected in this study from Chinese bamboo rats (Rhizomys sinensis) on nine farms in Jiangxi, Guangxi and Hainan provinces, China. They were analyzed for Cryptosporidium spp. using PCR and sequence analyses of the small subunit rRNA gene. Cryptosporidium parvum, C. parvum-like and C. ubiquitum-like genotypes identified were subtyped by sequence analysis of the 60 kDa glycoprotein (gp60) gene. RESULTS: Altogether, Cryptosporidium spp. were detected in 209 (29.5%) samples. The detection rate in samples from animals under two months of age (70.0%,105/150) was significantly higher than in samples from animals above 2 months (18.6%, 104/559; χ2 = 150.27, df = 1, P < 0.0001). Four Cryptosporidium species/genotypes were identified: C. parvum (n = 78); C. occultus (n = 1); a new genotype that is genetically related to C. ubiquitum (n = 85); and another new genotype that is genetically related to C. parvum (n = 44). Among them, C. parvum (27,610 ± 71,911 oocysts/gram of feces) and the C. parvum-like genotype (38,679 ± 82,811 oocysts/gram of feces) had higher oocyst shedding intensity than the C. ubiquitum-like genotype (2470 ± 7017 oocysts/gram of feces) and the C. occultus (1012 oocysts/gram of feces). The C. parvum identified belonged to three subtypes in two rare subtype families, including IIpA9 (n = 43), IIpA6 (n = 6) and IIoA15G1 (n = 9), while the C. parvum-like and C. ubiquitum-like genotypes generated very divergent gp60 sequences. CONCLUSIONS: Results of the present study suggest that bamboo rats on the study farms were infected with diverse Cryptosporidium species and divergent C. parvum subtypes, which probably had originated from their native habitats. As similar C. parvum subtypes have been recently detected in humans and farmed macaques, attentions should be paid to the potential role of these new farm animals in the transmission of zoonotic pathogens.

• Klíčová slova
• Bamboo rat, Cryptosporidium, Molecular epidemiology, Subtype, Zoonotic,
• MeSH
• Cryptosporidium parvum genetika   izolace a purifikace   MeSH
• Cryptosporidium genetika   izolace a purifikace   MeSH
• farmy * MeSH
• feces parazitologie   MeSH
• genotyp MeSH
• geny rRNA MeSH
• kryptosporidióza epidemiologie   parazitologie   MeSH
• lidé MeSH
• molekulární epidemiologie MeSH
• Muridae parazitologie   MeSH
• oocysty MeSH
• polymerázová řetězová reakce veterinární   MeSH
• protozoální DNA MeSH
• zoonózy diagnóza   parazitologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Čína epidemiologie   MeSH
• Názvy látek
• protozoální DNA MeSH

Fecal samples from wild-caught common voles (n = 328) from 16 locations in the Czech Republic were screened for Cryptosporidium by microscopy and PCR/sequencing at loci coding small-subunit rRNA, Cryptosporidium oocyst wall protein, actin and 70 kDa heat shock protein. Cryptosporidium infections were detected in 74 voles (22.6%). Rates of infection did not differ between males and females nor between juveniles and adults. Phylogenetic analysis revealed the presence of eight Cryptosporidium species/genotypes including two new species, C. alticolis and C. microti. These species from wild-caught common voles were able to infect common and meadow voles under experimental conditions, with a prepatent period of 3-5 days post-infection (DPI), but they were not infectious for various other rodents or chickens. Meadow voles lost infection earlier than common voles (11-14 vs 13-16 DPI) and had significantly lower infection intensity. Cryptosporidium alticolis infects the anterior small intestine and has larger oocysts (5.4 × 4.9 µm), whereas C. microti infects the large intestine and has smaller oocysts (4.3 × 4.1 µm). None of the rodents developed clinical signs of infection. Genetic and biological data support the establishment of C. alticolis and C. microti as separate species of the genus Cryptosporidium.

• Klíčová slova
• Experimental infection, Rodentia, molecular analyses, oocyst size, phylogeny, voles,
• MeSH
• Arvicolinae parazitologie   MeSH
• Cryptosporidium klasifikace   genetika   ultrastruktura   MeSH
• feces parazitologie   MeSH
• fluorescenční mikroskopie MeSH
• fylogeneze MeSH
• gastrointestinální trakt parazitologie   patologie   ultrastruktura   MeSH
• genetická variace MeSH
• interferenční mikroskopie MeSH
• kryptosporidióza epidemiologie   parazitologie   přenos   MeSH
• krysa rodu Rattus MeSH
• kur domácí MeSH
• mikroskopie elektronová rastrovací MeSH
• Murinae MeSH
• myši inbrední BALB C MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nemoci hlodavců epidemiologie   parazitologie   přenos   MeSH
• polymerázová řetězová reakce MeSH
• prevalence MeSH
• protozoální DNA chemie   genetika   izolace a purifikace   MeSH
• RNA ribozomální genetika   MeSH
• sekvence nukleotidů MeSH
• sekvenční seřazení veterinární   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• protozoální DNA MeSH
• RNA ribozomální MeSH