The Eurasian beaver (Castor fiber) is an expanding species in Europe in recent decades due to reintroductions and natural population growth. Beavers expanded rapidly in the second half of the 20th century, and their expansion was particularly rapid in the Danube basin. Nowadays, the majority of the continuous population located in the central and eastern parts of the continent and a large disjunct population in Norway and Sweden. Despite the increasing population size, the role of the beaver as a source of waterborne pathogens is not firmly established or is often inferred from circumstantial data. In order to extend knowledge about the composition of the parasite fauna of beavers occurring in Slovakia, 21 faecal samples taken near their burrows from three sites (located in the Topľa, Poprad and Danube river basin) were examined microscopically and by polymerase chain reaction (PCR). PCR-positive specimens were further examined by DNA sequencing. Parasites were detected in 21% of the examined beavers, specifically the protozoa Cryptosporidium spp. (n = 2), Blastocystis sp. (n = 1), and microsporidia Enterocytozoon bieneusi (n = 1) and Encephalitozoon spp. (n = 1). Using the sequence analysis, two variants of Cryptosporidium proliferans, a new subtype of Blastocystis sp., genotype D of E. bieneusi and Encephalitozoon intestinalis were identified. A putatively novel Blastocystis subtype (ST), originated from a site near the Danube river (southwestern Slovakia), was proposed based on high genetic divergence from the closest described subtype ST12 (11.9%) and unique phylogenetic position in a clade composed of ST's 35-38. The increased risk of zoonotic transmission or transmission to other animals was particularly evident in the site near the Topľa river (northeastern Slovakia), where fungal spores of zoonotic genotype D of E. bieneusi and E. intestinalis, together with oocysts of the potentially zoonotic C. proliferans, were found.
- Keywords
- Gastrointestinal parasites, Molecular analysis, Protected animals, Protozoan, Rodents, Zoonoses,
- Publication type
- Journal Article 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.
- Keywords
- Biology, Course of infection, Cryptosporidiosis, Genetic diversity, Mortality, Oocyst size, Phylogeny,
- MeSH
- Cryptosporidiidae * MeSH
- Cryptosporidium * MeSH
- Feces parasitology MeSH
- Ferrets MeSH
- Phylogeny MeSH
- Genotype MeSH
- Gerbillinae MeSH
- Cryptosporidiosis * parasitology MeSH
- Chickens MeSH
- Humans MeSH
- Guinea Pigs MeSH
- Mice MeSH
- Oocysts MeSH
- Sciuridae parasitology MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Guinea Pigs MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article 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.
- Keywords
- adaptation, biology, course of infection, infectivity, oocyst size, parasite, phylogeny, prevalence,
- 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
BACKGROUND: Cryptosporidium is an important gut microbe whose contributions towards infant and immunocompromise patient mortality rates are steadily increasing. Over the last decade, we have seen the development of various tools and methods for studying Cryptosporidium infection and its interactions with their hosts. One area that is sorely overlooked is the effect infection has on host metabolic processes. RESULTS: Using a 1H nuclear magnetic resonance approach to metabolomics, we have explored the nature of the mouse gut metabolome as well as providing the first insight into the metabolome of an infected cell line. Statistical analysis and predictive modelling demonstrated new understandings of the effects of a Cryptosporidium infection, while verifying the presence of known metabolic changes. Of note is the potential contribution of host derived taurine to the diarrhoeal aspects of the disease previously attributed to a solely parasite-based alteration of the gut environment, in addition to other metabolites involved with host cell catabolism. CONCLUSION: This approach will spearhead our understanding of the Cryptosporidium-host metabolic exchange and provide novel targets for tackling this deadly parasite.
- Keywords
- COLO-680N, Cryptosporidiosis, Metabolomics, NMR, Taurine,
- Publication type
- Journal Article MeSH
We undertook a study on Cryptosporidium spp. in wild cricetid rodents. Fecal samples were collected from meadow voles (Microtus pennsylvanicus), southern red-backed voles (Myodes gapperi), woodland voles (Microtus pinetorum), muskrats (Ondatra zibethicus) and Peromyscus spp. mice in North America, and from bank voles (Myodes glareolus) and common voles (Microtus arvalis) in Europe. Isolates were characterized by sequence and phylogenetic analyses of the small subunit ribosomal RNA (SSU) and actin genes. Overall, 33·2% (362/1089) of cricetids tested positive for Cryptosporidium, with a greater prevalence in cricetids from North America (50·7%; 302/596) than Europe (12·1%; 60/493). Principal Coordinate analysis separated SSU sequences into three major groups (G1-G3), each represented by sequences from North American and European cricetids. A maximum likelihood tree of SSU sequences had low bootstrap support and showed G1 to be more heterogeneous than G2 or G3. Actin and concatenated actin-SSU trees, which were better resolved and had higher bootstrap support than the SSU phylogeny, showed that closely related cricetid hosts in Europe and North America are infected with closely related Cryptosporidium genotypes. Cricetids were not major reservoirs of human pathogenic Cryptosporidium spp.
- Keywords
- Cryptosporidium, Cricetidae, biogeography, phylogenetics,
- MeSH
- Arvicolinae parasitology MeSH
- Cryptosporidium classification isolation & purification pathogenicity physiology MeSH
- Animals, Wild parasitology MeSH
- Feces parasitology MeSH
- Phylogeny MeSH
- Phylogeography MeSH
- Genotype MeSH
- Rodentia parasitology MeSH
- Cryptosporidiosis epidemiology parasitology MeSH
- Mice parasitology MeSH
- RNA, Ribosomal genetics MeSH
- Sequence Analysis, DNA MeSH
- Disease Reservoirs parasitology MeSH
- Animals MeSH
- Check Tag
- Mice parasitology MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Europe epidemiology MeSH
- North America epidemiology MeSH
- Names of Substances
- RNA, Ribosomal 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.
- Keywords
- One Health, cryptosporidiosis, giardiasis, zoonosis,
- MeSH
- Cryptosporidium genetics isolation & purification MeSH
- Feces parasitology MeSH
- Giardia genetics isolation & purification MeSH
- Giardiasis epidemiology parasitology MeSH
- Cryptosporidiosis epidemiology parasitology MeSH
- Real-Time Polymerase Chain Reaction MeSH
- Humans MeSH
- Foodborne Diseases epidemiology parasitology MeSH
- Prevalence MeSH
- Public Health * MeSH
- Environment MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Review MeSH
- Geographicals
- Europe, Eastern epidemiology MeSH
