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: Avian cryptosporidiosis is a common parasitic disease that is caused by five species, which are well characterised at the molecular and biological level, and more than 18 genotypes for which we have limited information. In this study, we determined the occurrence and molecular characteristics of Cryptosporidium spp. in farmed ostriches in the Czech Republic. METHODS: The occurrence and genetic identity of Cryptosporidium spp. were analysed by microscopy and PCR/sequencing of the small subunit rRNA, actin, HSP70 and gp60 genes. Cryptosporidium avian genotype II was examined from naturally and experimentally infected hosts and measured using differential interference contrast. The localisation of the life-cycle stages was studied by electron microscopy and histologically. Infectivity of Cryptosporidium avian genotype II for cockatiels (Nymphicus hollandicus (Kerr)), chickens (Gallus gallus f. domestica (L.)), geese (Anser anser f. domestica (L.)), SCID and BALB/c mice (Mus musculus L.) was verified. RESULTS: A total of 204 individual faecal samples were examined for Cryptosporidium spp. using differential staining and PCR/sequencing. Phylogenetic analysis of small subunit rRNA, actin, HSP70 and gp60 gene sequences showed the presence of Cryptosporidium avian genotype II (n = 7) and C. ubiquitum Fayer, Santín & Macarisin, 2010 IXa (n = 5). Only ostriches infected with Cryptosporidium avian genotype II shed oocysts that were detectable by microscopy. Oocysts were purified from a pooled sample of four birds, characterised morphometrically and used in experimental infections to determine biological characteristics. Oocysts of Cryptosporidium avian genotype II measure on average 6.13 × 5.15 μm, and are indistinguishable by size from C. baileyi Current, Upton & Haynes, 1986 and C. avium Holubová, Sak, Horčičková, Hlásková, Květoňová, Menchaca, McEvoy & Kváč, 2016. Cryptosporidium avian genotype II was experimentally infectious for geese, chickens and cockatiels, with a prepatent period of four, seven and eight days post-infection, respectively. The infection intensity ranged from 1000 to 16,000 oocysts per gram. None of the naturally or experimentally infected birds developed clinical signs in the present study. CONCLUSIONS: The molecular and biological characteristics of Cryptosporidium avian genotype II, described here, support the establishment of a new species, Cryptosporidium ornithophilus n. sp.

• Klíčová slova
• C. ubiquitum, Cryptosporidium avian genotype II, Cryptosporidium ornithophilus n. sp., Experimental infections, Occurrence, Oocyst size, PCR,
• MeSH
• Cryptosporidium klasifikace   genetika   ultrastruktura   MeSH
• fylogeneze MeSH
• hospodářská zvířata parazitologie   MeSH
• hostitelská specificita MeSH
• klasifikace MeSH
• kryptosporidióza parazitologie   MeSH
• nemoci ptáků parazitologie   MeSH
• protozoální geny genetika   MeSH
• ptáci parazitologie   MeSH
• stadia vývoje MeSH
• Struthioniformes parazitologie   MeSH
• taxonomické DNA čárové kódování veterinární   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články 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 muris strain TS03 are described, and the species name Cryptosporidium proliferans n. sp. is proposed. Cryptosporidium proliferans obtained from a naturally infected East African mole rat (Tachyoryctes splendens) in Kenya was propagated under laboratory conditions in rodents (SCID mice and southern multimammate mice, Mastomys coucha) and used in experiments to examine oocyst morphology and transmission. DNA from the propagated C. proliferans isolate, and C. proliferans DNA isolated from the feces of an African buffalo (Syncerus caffer) in Central African Republic, a donkey (Equus africanus) in Algeria, and a domestic horse (Equus caballus) in the Czech Republic were used for phylogenetic analyses. Oocysts of C. proliferans are morphologically distinguishable from C. parvum and C. muris HZ206, measuring 6.8-8.8 (mean = 7.7 μm) × 4.8-6.2 μm (mean = 5.3) with a length to width ratio of 1.48 (n = 100). Experimental studies using an isolate originated from T. splendens have shown that the course of C. proliferans infection in rodent hosts differs from that of C. muris and C. andersoni. The prepatent period of 18-21 days post infection (DPI) for C. proliferans in southern multimammate mice (Mastomys coucha) was similar to that of C. andersoni and longer than the 6-8 DPI prepatent period for C. muris RN66 and HZ206 in the same host. Histopatologicaly, stomach glands of southern multimammate mice infected with C. proliferans were markedly dilated and filled with necrotic material, mucus, and numerous Cryptosporidium developmental stages. Epithelial cells of infected glands were atrophic, exhibited cuboidal or squamous metaplasia, and significantly proliferated into the lumen of the stomach, forming papillary structures. The epithelial height and stomach weight were six-fold greater than in non-infected controls. Phylogenetic analyses based on small subunit rRNA, Cryptosporidium oocyst wall protein, thrombospondin-related adhesive protein of Cryptosporidium-1, heat shock protein 70, actin, heat shock protein 90 (MS2), MS1, MS3, and M16 gene sequences revealed that C. proliferans is genetically distinct from C. muris and other previously described Cryptosporidium species.

• MeSH
• Cryptosporidium klasifikace   genetika   MeSH
• fylogeneze MeSH
• kryptosporidióza parazitologie   MeSH
• mikroftalmičtí podzemní hlodavci MeSH
• myši SCID MeSH
• myši MeSH
• oocysty metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

From 2011 to 2012, to identify Cryptosporidium spp. occurrence in Eurasian wild boars (Sus scrofa) 29 randomly selected localities (both forest areas and enclosures) across the Central European countries of Austria, the Czech Republic, Poland, and the Slovak Republic were investigated. Cryptosporidium oocysts were microscopicaly detected in 11 out of 460 faecal samples examined using aniline-carbol-methyl violet staining. Sixty-one Cryptosporidium infections, including the 11 infections that were detected by microscopy, were detected using genus- or species-specific nested PCR amplification of SSU rDNA. This represents a 5.5 fold greater sensitivity for PCR relative to microscopy. Combining genus- and species-specific PCR tools significantly changes the perspective on the occurrence of Cryptosporidium spp. in wild boars. While RFLP and direct sequencing of genus specific PCR-amplified products revealed 56 C. suis (20) and C. scrofarum (36) monoinfections and only 5 mixed infections of these species, species-specific molecular tools showed 44 monoinfections and 17 mixed infections with these species. PCR analysis of the gp60 gene did not reveal any other Cryptosporidium infections. Similar to domestic pigs, C. scrofarum was detected as a dominant species infecting adult Eurasian wild boars (Sus scrofa). Cryptosporidium infected wild boars did not show signs of clinical disease. This report is perhaps the most comprehensive survey of cryptosporidial infection in wild boars.

• Klíčová slova
• Central Europe, Cryptosporidium scrofarum, Cryptosporidium suis, Eurasian wild boar, PCR, SSU,
• MeSH
• Cryptosporidium klasifikace   MeSH
• druhová specificita MeSH
• feces parazitologie   MeSH
• kryptosporidióza epidemiologie   veterinární   MeSH
• nemoci prasat epidemiologie   parazitologie   MeSH
• prasata MeSH
• Sus scrofa parazitologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Geografické názvy
• Evropa epidemiologie   MeSH

From 2009 to 2011, the occurrence of Cryptosporidium spp. was investigated on 22 farms in the Czech Republic. A total of 1,620 individual faecal samples of pigs of all age categories (pre-weaned, starters, pre-growers, growers, and sows) were evaluated for presence of Cryptosporidium spp. by standard microscopy and molecular tools. Genotyping was done through PCR amplification and characterization of the SSU rRNA (species-specific protocols) and GP60 loci. Cryptosporidium spp. was found on 16 of 22 farms with a range 0.9-71.4 %. Overall, 194 (12 %) specimens were positive by microscopy and 353 (21.8 %) by PCR. While RFLP and direct sequencing of the PCR-amplified products showed presence of Cryptosporidium suis (142), Cryptosporidium scrofarum (195), Cryptosporidium muris (3) and 13 samples had mixed infections with C. suis and C. scrofarum, species-specific molecular tools identified C. suis (224), C. scrofarum (208), Cryptosporidium parvum subtype IIa A16G1R1b (1), and C. muris (3). In addition, a total of 82 pigs had concurrent infections with C. suis and C. scrofarum. The analysis by age showed that C. suis was primarily detected among pre-weaned, whereas C. scrofarum was mostly detected among starters, especially those weaned at a younger age. Moreover, C. scrofarum never has been detected in animals younger than 6 weeks of age. Also, piglets weaned at 3 weeks of age were twice more likely to be infected with C. scrofarum than piglets weaned at an older age. Pigs raised on straw bedding were more likely to have Cryptosporidium than pigs raised on slats/slurry systems. The infections with different species were not associated with loose faeces or intensity of oocyst shedding, even when comparing different age groups.

• MeSH
• chov zvířat metody   MeSH
• Cryptosporidium klasifikace   cytologie   genetika   izolace a purifikace   MeSH
• feces parazitologie   MeSH
• genotyp MeSH
• kryptosporidióza epidemiologie   parazitologie   veterinární   MeSH
• mikroskopie MeSH
• nemoci prasat epidemiologie   parazitologie   MeSH
• polymerázová řetězová reakce MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• prasata MeSH
• prevalence MeSH
• věkové faktory MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika epidemiologie   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