Cryptosporidium spp. are common protozoan pathogens in mammals. The diversity and biology of Cryptosporidium in tree squirrels are not well studied. A total of 258 Eurasian red squirrels (Sciurus vulgaris) from 25 and 15 locations in the Czech Republic and Slovakia, respectively, were examined for Cryptosporidium spp. oocysts and specific DNA at the SSU, actin, HSP70, TRAP-C1, COWP, and gp60 loci. Out of 26 positive animals, only juveniles (9/12) were microscopically positive (18,000 to 72,000 OPG), and molecular analyses revealed the presence of Cryptosporidium sp. ferret genotype in all specimens. Oocysts obtained from naturally-infected squirrels measured 5.54-5.22 μm and were not infectious for laboratory mice (BALB/c and SCID), Mongolian gerbils, Guinea pigs, Southern multimammate mice, chickens, or budgerigars. None of naturally infected squirrels showed clinical signs of disease. The frequency of occurrence of the ferret genotype in squirrels did not vary statistically based on host age, gender or country of capture. Phylogenetic analysis of sequences from six loci revealed that Cryptosporidium sp. ferret genotype is genetically distinct from the currently accepted Cryptosporidium species. Morphological and biological data from this and previous studies support the establishment of Cryptosporidium sp. ferret genotype as a new species, Cryptosporidium sciurinum n. sp.

• Keywords
• Cryptosporidium sp. ferret genotype, biology, course of infection, infectivity, occurrence, oocyst size, phylogeny,
• Publication type
• Journal Article 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.

• Keywords
• Cryptosporidium ratti, infectivity, morphometric analysis, phylogeny, prevalence,
• MeSH
• Actins genetics   MeSH
• Cryptosporidium * classification   genetics   isolation & purification   MeSH
• Animals, Wild parasitology   MeSH
• Feces parasitology   MeSH
• Phylogeny MeSH
• Genetic Variation MeSH
• Classification MeSH
• Rats parasitology   MeSH
• Mice MeSH
• Prevalence MeSH
• HSP70 Heat-Shock Proteins genetics   MeSH
• DNA, Protozoan MeSH
• DNA, Ribosomal genetics   MeSH
• Animals MeSH
• Check Tag
• Rats parasitology   MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Actins MeSH
• HSP70 Heat-Shock Proteins MeSH
• DNA, Protozoan MeSH
• DNA, Ribosomal MeSH

The genetic diversity of Cryptosporidium spp. in Apodemus spp. (striped field mouse, yellow-necked mouse and wood mouse) from 16 European countries was examined by PCR/sequencing of isolates from 437 animals. Overall, 13.7% (60/437) of animals were positive for Cryptosporidium by PCR. Phylogenetic analysis of small-subunit rRNA, Cryptosporidium oocyst wall protein and actin gene sequences showed the presence of Cryptosporidium ditrichi (22/60), Cryptosporidium apodemi (13/60), Cryptosporidium apodemus genotype I (8/60), Cryptosporidium apodemus genotype II (9/60), Cryptosporidium parvum (2/60), Cryptosporidium microti (2/60), Cryptosporidium muris (2/60) and Cryptosporidium tyzzeri (2/60). At the gp60 locus, novel gp60 families XVIIa and XVIIIa were identified in Cryptosporidium apodemus genotype I and II, respectively, subtype IIaA16G1R1b was identified in C. parvum, and subtypes IXaA8 and IXcA6 in C. tyzzeri. Only animals infected with C. ditrichi, C. apodemi, and Cryptosporidium apodemus genotypes shed oocysts that were detectable by microscopy, with the infection intensity ranging from 2000 to 52,000 oocysts per gram of faeces. None of the faecal samples was diarrheic in the time of the sampling.

• Keywords
• Epidemiology, Molecular analyses, Phylogeny, Rodentia,
• MeSH
• Cryptosporidium genetics   MeSH
• Genetic Variation * MeSH
• Genotype MeSH
• Cryptosporidiosis parasitology   MeSH
• Murinae microbiology   MeSH
• Mice MeSH
• RNA, Ribosomal, 18S genetics   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Europe MeSH
• Names of Substances
• RNA, Ribosomal, 18S 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 classification   genetics   MeSH
• Phylogeny MeSH
• Cryptosporidiosis parasitology   MeSH
• Mole Rats MeSH
• Mice, SCID MeSH
• Mice MeSH
• Oocysts metabolism   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't 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.

• Keywords
• Central Europe, Cryptosporidium scrofarum, Cryptosporidium suis, Eurasian wild boar, PCR, SSU,
• MeSH
• Cryptosporidium classification   MeSH
• Species Specificity MeSH
• Feces parasitology   MeSH
• Cryptosporidiosis epidemiology   veterinary   MeSH
• Swine Diseases epidemiology   parasitology   MeSH
• Swine MeSH
• Sus scrofa parasitology   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH
• Geographicals
• Europe epidemiology   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
• Animal Husbandry methods   MeSH
• Cryptosporidium classification   cytology   genetics   isolation & purification   MeSH
• Feces parasitology   MeSH
• Genotype MeSH
• Cryptosporidiosis epidemiology   parasitology   veterinary   MeSH
• Microscopy MeSH
• Swine Diseases epidemiology   parasitology   MeSH
• Polymerase Chain Reaction MeSH
• Polymorphism, Restriction Fragment Length MeSH
• Swine MeSH
• Prevalence MeSH
• Age Factors MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Czech Republic epidemiology   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 classification   cytology   genetics   MeSH
• Species Specificity MeSH
• Feces parasitology   MeSH
• Phylogeny * MeSH
• Genotype MeSH
• Gerbillinae MeSH
• Cryptosporidiosis pathology   veterinary   MeSH
• Guinea Pigs MeSH
• Mice, Inbred BALB C MeSH
• Mice, SCID MeSH
• Mice MeSH
• Swine Diseases parasitology   pathology   MeSH
• Swine MeSH
• Genes, Protozoan genetics   MeSH
• Animals MeSH
• Check Tag
• Guinea Pigs MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH