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.

• Klíčová slova
• Gastrointestinal parasites, Molecular analysis, Protected animals, Protozoan, Rodents, Zoonoses,
• Publikační typ
• časopisecké články MeSH

PURPOSE: Parasites of genus Encephalitozoon are well known pathogens of domestic animals however less attention was paid to its spread among wildlife that can play an important role of reservoir of infection. The aim of the study was to conduct molecular detection and genotype characterization of Encephalitozoon spp. in wild small mammals trapped in localities both near to and at a large distance from residential areas. METHODS: In total, 300 wild small mammals (274 Rodentia and 26 Eulipotyphla) were trapped in 41 localities of the Czech Republic and tested by nested PCR for Encephalitozoon spp. RESULTS: The DNA of Encephalitozoon spp. was proved in tissues (brain or liver) of 11% (32/300) of animals. There was a statistically significant difference (p < 0.001) in positivity among animal species with the most infected species Micromys minutus (50%, 4/8) and Myodes glareolus (17%, 9/53). There was also statistically significant difference (p < 0.001) between localities with the higher positivity (29%, 12/42) in localities near to residential areas, compared to localities with a large distance from residential areas (8%, 20/258). Sex and age of wild small mammals did not have effect on their positivity. Genotyping analysis revealed E. cuniculi genotype II in 22 samples and E. hellem genotype 1 A in one sample. CONCLUSION: This study brings new information on the molecular characterization of Encephalitozoon spp. isolated from wild small mammals trapped in two different areas (localities in near to residential areas and localities with a large distance from residential areas).

• Klíčová slova
• Encephalitozoonosis, Genotyping, Urban area, Wildlife, Zoonosis,
• MeSH
• divoká zvířata * parazitologie   MeSH
• Encephalitozoon cuniculi genetika   izolace a purifikace   klasifikace   MeSH
• Encephalitozoon * genetika   izolace a purifikace   klasifikace   MeSH
• encephalitozoonóza * veterinární   epidemiologie   parazitologie   mikrobiologie   MeSH
• genotyp * MeSH
• hlodavci * parazitologie   MeSH
• hmyzožravci parazitologie   MeSH
• játra parazitologie   MeSH
• mozek parazitologie   MeSH
• polymerázová řetězová reakce MeSH
• zdroje nemoci * parazitologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

We detected 24 Encephalitozoon cuniculi positive Ixodes ricinus ticks of 284 collected in the Czech Republic. Since the route of transmission of microsporidia is not fully understood, the presence of microsporidia in ticks raises the question of whether they may be involved in the transmission of these pathogens.

• Klíčová slova
• Encephalitozoon cuniculi, Ixodes ricinus, PCR, qPCR, vector,
• MeSH
• Encephalitozoon cuniculi izolace a purifikace   genetika   MeSH
• encephalitozoonóza přenos   mikrobiologie   MeSH
• klíště * mikrobiologie   MeSH
• lidé MeSH
• mikrosporidióza přenos   MeSH
• nemoci přenášené klíšťaty přenos   mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

We retrospectively analyzed of 211 frozen cerebrospinal fluid samples from immunocompetent persons in the Czech Republic and detected 6 Encephalitozoon cuniculi-positive samples. Microsporidiosis is generally underestimated and patients are not usually tested for microsporidia, but latent infection in immunodeficient and immunocompetent patients can cause serious complications if not detected and treated.

• Klíčová slova
• Czech Republic, Encephalitozoon cuniculi, PCR, cerebrospinal fluid, latent infection, parasites, qPCR, zoonoses,
• MeSH
• dospělí MeSH
• Encephalitozoon cuniculi * izolace a purifikace   genetika   MeSH
• encephalitozoonóza * mozkomíšní mok   mikrobiologie   epidemiologie   MeSH
• imunokompetence MeSH
• lidé středního věku MeSH
• lidé MeSH
• retrospektivní studie MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH

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

Cryptosporidium spp., Enterocytozoon bieneusi and Encephalitozoon spp. are the most common protistan parasites of vertebrates. The results show that pigeon populations in Central Europe are parasitised by different species of Cryptosporidium and genotypes of microsporidia of the genera Enterocytozoon and Encephalitozoon. A total of 634 and 306 faecal samples of captive and feral pigeons (Columba livia f. domestica) from 44 locations in the Czech Republic, Slovakia and Poland were analysed for the presence of parasites by microscopy and PCR/sequence analysis of small subunit ribosomal RNA (18S rDNA), 60 kDa glycoprotein (gp60) and internal transcribed spacer (ITS) of SSU rDNA. Phylogenetic analyses revealed the presence of C. meleagridis, C. baileyi, C. parvum, C. andersoni, C. muris, C. galli and C. ornithophilus, E. hellem genotype 1A and 2B, E. cuniculi genotype I and II and E. bieneusi genotype Peru 6, CHN-F1, D, Peru 8, Type IV, ZY37, E, CHN4, SCF2 and WR4. Captive pigeons were significantly more frequently parasitised with screened parasite than feral pigeons. Cryptosporidium meleagridis IIIa and a new subtype IIIl have been described, the oocysts of which are not infectious to immunodeficient mice, whereas chickens are susceptible. This investigation demonstrates that pigeons can be hosts to numerous species, genotypes and subtypes of the studied parasites. Consequently, they represent a potential source of infection for both livestock and humans.

• Klíčová slova
• Birds, Experimental infection, Genotyping, PCR,
• MeSH
• Columbidae MeSH
• Cryptosporidium * genetika   MeSH
• Encephalitozoon * genetika   MeSH
• Enterocytozoon * genetika   MeSH
• feces parazitologie   MeSH
• fylogeneze MeSH
• genetická variace MeSH
• genotyp MeSH
• kryptosporidióza * epidemiologie   parazitologie   MeSH
• kur domácí MeSH
• lidé MeSH
• mikrosporidióza * epidemiologie   veterinární   parazitologie   MeSH
• myši MeSH
• ribozomální DNA MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa epidemiologie   MeSH
• Názvy látek
• ribozomální DNA MeSH

Bats may carry various viruses and bacteria which can be harmful to humans, but little is known about their role as a parasitic source with zoonotic potential. The aim of this study was to test wild bats for the presence of selected parasites: Toxoplasma gondii, Neospora caninum and microsporidia Encephalitozoon spp. In total, brain and small intestine tissues of 100 bats (52 Myotis myotis, 43 Nyctalus noctula and 5 Vespertilio murinus) were used for the DNA isolation and PCR detection of the abovementioned agents. Toxoplasma gondii DNA was detected by real-time PCR in 1% of bats (in one male of M. myotis), while all bats were negative for N. caninum DNA. Encephalitozoon spp. DNA was detected by nested PCR in 25% of bats, including three species (twenty-two M. myotis, two N. noctula and one V. murinus). Positive samples were sequenced and showed homology with the genotypes Encephalitozoon cuniculi II and Encephalitozoon hellem 2C. This is the first study on wild vespertilionid bats from Central Europe and worldwide, with a relatively high positivity of Encephalitozoon spp. detected in bats.

• Klíčová slova
• insectivore, microsporidiosis, molecular methods, neosporosis, toxoplasmosis,
• MeSH
• Chiroptera * MeSH
• Encephalitozoon * genetika   MeSH
• kokcidióza * veterinární   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé MeSH
• Neospora * genetika   MeSH
• paraziti * genetika   MeSH
• Toxoplasma * genetika   MeSH
• toxoplazmóza zvířat * parazitologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa MeSH

Microsporidia are pathogenic organism related to fungi. They cause infections in a wide variety of mammals as well as in avian, amphibian, and reptilian hosts. Many microsporidia species play an important role in the development of serious diseases that have significant implications in human and veterinary medicine. While microsporidia were originally considered to be opportunistic pathogens in humans, it is now understood that infections also occur in immune competent humans. Encephalitozoon cuniculi, Encephalitozoon intestinalis, and Enterocytozoon bieneusi are primarily mammalian pathogens. However, many other species of microsporidia that have some other primary host that is not a mammal have been reported to cause sporadic mammalian infections. Experimental models and observations in natural infections have demonstrated that microsporidia can cause a latent infection in mammalian hosts. This chapter reviews the published studies on mammalian microsporidiosis and the data on chronic infections due to these enigmatic pathogens.

• Klíčová slova
• Epidemiology, Infection, Latency, Mammals, Microsporidia, Recurrent infection, Transmission,
• MeSH
• Enterocytozoon * MeSH
• feces mikrobiologie   MeSH
• lidé MeSH
• Microsporidia * genetika   MeSH
• perzistentní infekce MeSH
• savci MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Microsporidia of the genus Encephalitozoon are usually associated with severe infections in immunodeficient hosts while, in immunocompetent ones, microsporidiosis produces minimal clinically apparent disease. Despite their microscopic size, microsporidia are capable of causing systemic infection within a few days. However, the mechanisms by which microsporidia reach target tissues during acute infection remain unclear. Out of four genotypes of Encephalitozoon cuniculi, only three are available for experimental studies, with E. cuniculi genotype II being the best characterized. METHODS: In the present study, we tested the association between inflammation induction in immunocompetent and immunodeficient mice and the presence of spores of E. cuniculi genotypes I and III in selected organs using molecular methods and compared the results with previously published data on E. cuniculi genotype II. RESULTS: We reported the positive connection between inflammation induction and the significant increase of E. cuniculi genotypes I and III occurrence in inflammatory foci in both immunocompetent BALB/c and immunodeficient severe combined immunodeficient (SCID) mice in the acute phase of infection. The induction of inflammation resulted in increased concentration of E. cuniculi of both genotypes in the site of inflammation, as previously reported for E. cuniculi genotype II. Moreover, our study extended the spectrum of differences among E. cuniculi genotypes by the variations in dispersal rate within host bodies after experimentally induced inflammation. CONCLUSION: The results imply possible involvement of immune cells serving as vehicles transporting E. cuniculi towards inflammation foci. The elucidation of possible connection with pro-inflammatory immune responses represents an important challenge with implications for human health and the development of therapeutic strategies.

• Klíčová slova
• Encephalitozoon cuniculi genotype I, Encephalitozoon cuniculi genotype III, inflammation, targeted migration,
• Publikační typ
• časopisecké články MeSH

Objectives: Patients with inflammatory bowel disease (IBD) are susceptible to intestinal opportunistic infections due to both defective mucosal immunity and altered immune response resulting from immunosuppressive treatment. Microsporidia infecting the gastrointestinal tract and causing diarrhoea can potentially affect the course of IBD. Methods: Stool samples (90 IBD children and 121 healthy age-matched controls) were screened for Encephalitozoon spp. and Enterocytozoon bieneusi by microscopy and polymerase chain reaction followed by sequencing. Results: E. bieneusi genotype D was found in seven out of 90 (7.8%) IBD children. No children from the control group were infected, making the pathogen prevalence in the IBD group significant (P = 0.002). Furthermore, infection was confirmed only in patients receiving immunosuppressive treatment (P = 0.013). Conclusions: Children with IBD are at risk of intestinal E. bieneusi infection, especially when receiving immunosuppressive treatment. Therefore, microsporidia should be considered as a significant infectious agent in this group of patients.

• Klíčová slova
• Enterocytozoon bieneusi, children, immunosuppressive treatment, inflammatory bowel disease, molecular characterization,
• Publikační typ
• časopisecké články MeSH