The domestic chinchilla (Chinchilla lanigera) is kept as a pet and previous studies suggest that it may play an important role as a source of zoonotic parasites, including Giardia intestinalis, Cryptosporidium spp. and microsporidia. In this study, we examined the occurrence and genetic diversity of above mentioned parasites in pet chinchillas in the Czech Republic by PCR/sequencing of the 18S rRNA, TPI, and ITS genes. Of 149 chinchillas from 24 breeders, 91.3 % were positive for G. intestinalis, 8.1 % for Cryptosporidium spp., 2.0 % for Encephalitozoon spp., and 5.4 % for E. bieneusi. Molecular analyses revealed presence of G. intestinalis assemblage B, C. ubiquitum (XIIa family), E. bieneusi genotypes D, SCF2, and, CHN-F1, and E. intestinalis. The infection intensity of G. intestinalis determined by qRT-PCR reached up to 53,978 CPG, C. ubiquitum up to 1409 OPG, E. intestinalis up to 1124 SPG, and E. bieneusi up to 1373 SPG. Only two chinchillas with C. ubiquitum and five with G. intestinalis had diarrhoea at the time of the screening. Three chinchillas in the long-term study were consistently positive for G. intestinalis, with intermittent excretion of C. ubiquitum, E. intestinalis, and E. bieneusi over 25 weeks. The findings indicate that chinchillas are frequently infected with zoonotic parasitic protists, but that these infections rarely show clinical signs. The lack of visible signs could reduce the vigilance of pet owners when handling their chinchillas, increasing the risk of transmission within breeding groups and possibly to humans.

• Klíčová slova
• PCR, Pets, QPCR, Zoonosis,
• MeSH
• činčila * parazitologie   MeSH
• Cryptosporidium * genetika   klasifikace   izolace a purifikace   MeSH
• domácí zvířata * MeSH
• Encephalitozoon * genetika   izolace a purifikace   klasifikace   MeSH
• encephalitozoonóza * veterinární   epidemiologie   mikrobiologie   MeSH
• Enterocytozoon * genetika   izolace a purifikace   MeSH
• feces parazitologie   mikrobiologie   MeSH
• Giardia lamblia * genetika   izolace a purifikace   klasifikace   MeSH
• giardiáza * veterinární   parazitologie   MeSH
• kryptosporidióza parazitologie   epidemiologie   MeSH
• mikrosporidióza * veterinární   epidemiologie   MeSH
• RNA ribozomální 18S genetika   MeSH
• zoonózy * parazitologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Názvy látek
• RNA ribozomální 18S 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

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

OBJECTIVE: In recent years new infectious diseases, i.e. emerging or re-emerging diseases, have been coming to the forefront. Currently, microsporidia, considered to be a major cause of emerging and opportunistic infections particularly in immunocompromised individuals, are also included in this group. Therefore, the aim of our study was to map the prevalence of Encephalitozoon intestinalis and Enterocytozoon bieneusi infection in a group of patients and to compare it with the occurrence of specific antigens in immunocompetent people. METHODS: Detection of spores of both pathogens in faecal samples was performed by an immunofluorescence test using species-specific monoclonal antibodies. RESULTS: Positivity to E. intestinalis in 91 examined immunosuppressed patients reached 33% (30/91), while only 4.3% (3/70) of the control group samples were found to be positive (relative risk 7.7, p < 0.001). In case of E. bieneusi 14.3% (13/91) of immunocompromised patients were positive, as were 5.7% (4/70) of people from the control group (relative risk 2.5, p = 0.095). CONCLUSION: In case of development of any opportunistic infection, the infection is detected and removed in most cases at an early stage. The incidence of clinically manifested microsporidiosis in patients with immunodeficiency is rare as they are under constant medical supervision. However, we must not forget about opportunistic infections, and in case of any non-specific symptoms it is necessary to exclude or confirm the diagnosis for immediate treatment.

• Klíčová slova
• Encephalitozoon intestinalis, Enterocytozoon bieneusi, immunocompromised patients, microsporidia, monoclonal antibodies,
• MeSH
• Encephalitozoon izolace a purifikace   MeSH
• encephalitozoonóza diagnóza   epidemiologie   MeSH
• Enterocytozoon izolace a purifikace   MeSH
• feces mikrobiologie   MeSH
• imunokompromitovaný pacient * MeSH
• lidé MeSH
• mikrosporidióza diagnóza   epidemiologie   MeSH
• oportunní infekce diagnóza   mikrobiologie   MeSH
• plošný screening * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Slovenská republika epidemiologie   MeSH

Microsporidia are emerging pathogens which cause an opportunistic infections in immunocompromised patients, especially those with AIDS. Intestinal microsporidiosis is the most recognized infection, whereas urinary tract infections caused by microsporidia are rarely paid attention to either due to their subclinical course or diagnostic difficulties. In this report dual microsporidial infection of urinary tract, caused by Enterocytozoon bieneusi and Encephalitozoon cuniculi was described in HIV/AIDS patients under cART therapy. Since microsporidiosis can cause severe complications or even death in immunosuppressed patients, our results suggest that microsporidial infection should be included in routine investigation of HIV-positive patients, even asymptomatic.

• MeSH
• Encephalitozoon cuniculi * fyziologie   MeSH
• encephalitozoonóza * komplikace   MeSH
• Enterocytozoon * fyziologie   MeSH
• HIV infekce komplikace   mikrobiologie   MeSH
• koinfekce MeSH
• lidé MeSH
• mikrosporidióza * komplikace   MeSH
• močové ústrojí * mikrobiologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

OBJECTIVES: Encephalitozoon spp. and Enterocytozoon bieneusi are intracellular parasitic fungi from the phylum Microsporidia, which initially localize to the intestine. As opportunistic pathogens, Encephalitozoon spp. in particular can disseminate to the respiratory tract, among other locations. Patients on life-long immunosuppression are at higher risk of such infections, mostly symptomatic. METHODS: Sputum samples and bronchial washings from 72 renal transplant recipients and 105 patients with various respiratory diseases were screened for Encephalitozoon spp. and E. bieneusi by microscopic examination and genus-specific nested PCR followed by genotyping. RESULTS: A total of 8.3% (6/72) of immunosuppressed renal transplant recipients and 1.9% (2/105) of patients with various respiratory diseases, both immunocompetent and immunosuppressed, were positive for respiratory microsporidial infection. All six transplant recipients were Encephalitozoon cuniculi-positive by PCR/sequencing and five of them suffered from respiratory symptoms. The presence of microsporidial spores was also confirmed microscopically in three of the transplant recipients. Of the two immunocompetent patients with various respiratory diseases, one had an E. cuniculi infection, while the second had an E. bieneusi infection. CONCLUSIONS: Life-long immunosuppression in renal transplant recipients increases the risk of respiratory infection by E. cuniculi. Microsporidia should be screened in respiratory samples of these patients, particularly when they have respiratory symptoms.

• Klíčová slova
• Encephalitozoon cuniculi, Enterocytozoon bieneusi, Life-long immunosuppression, Renal transplant recipients, Respiratory tract,
• MeSH
• dospělí MeSH
• Encephalitozoon cuniculi * genetika   izolace a purifikace   MeSH
• encephalitozoonóza mikrobiologie   MeSH
• Enterocytozoon genetika   izolace a purifikace   MeSH
• imunokompromitovaný pacient * MeSH
• infekce dýchací soustavy mikrobiologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• příjemce transplantátu MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• transplantace ledvin * MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi are common enteric pathogens in humans and animals. Data on the transmission of these pathogens are scarce from Guangdong, China, which has a subtropical monsoon climate and is the epicenter for many emerging infectious diseases. This study was conducted to better understand the prevalence and identity of the three pathogens in pre-weaned dairy calves in Guangdong. METHODS: The occurrence and genetic identity of three pathogens were analyzed by polymerase chain reaction. PCR-positive products were sequenced to determine the species and genotypes. A Chi-square test was used to compare the prevalence of pathogens among sampling dates, age groups, or clinical signs. RESULTS: The detection rates of Cryptosporidium spp., G. duodenalis and E. bieneusi were 24.0% (93/388), 74.2% (288/388) and 15.7% (61/388), respectively. Three Cryptosporidium species were detected, including C. bovis (n = 73), C. parvum (n = 12) and C. ryanae (n = 7); one animal had concurrence of C. bovis and C. parvum. C. parvum was the dominant species during the first two weeks of life, whereas C. bovis and C. ryanae were mostly seen at 3-9 weeks of age. Sequence analysis identified the C. parvum as subtype IIdA19G1. Assemblage E (n = 282), assemblage A (n = 1), and concurrence of A and E (n = 5) were identified among G. duodenalis-positive animals using multilocus genotyping (MLG). Altogether, 15, 10 and 17 subtypes of assemblage E were observed at the bg, gdh and tpi loci, respectively, forming 49 assemblage E MLGs. The highest detection rate of G. duodenalis was found in winter. Sequence analysis identified genotypes J (n = 57), D (n = 3) and one concurrence of J and D among E. bieneusi-positive animals. The detection rate of E. bieneusi was significantly higher in spring (38.0%; 41/108) than in summer (7.2%; 8/111) and winter (7.1%; 12/169). CONCLUSIONS: These results indicate a common occurrence of C. parvum subtype IIdA19G1, G. duodenalis assemblage E, and E. bieneusi genotype J in pre-weaned dairy calves in Guangdong. More studies are needed to understand the unique genetic characteristics and zoonotic potential of the three enteric pathogens in the province.

• Klíčová slova
• Cryptosporidium, Enterocytozoon bieneusi, Giardia duodenalis, Molecular epidemiology,
• MeSH
• Cryptosporidium genetika   izolace a purifikace   MeSH
• Enterocytozoon genetika   imunologie   MeSH
• genotyp MeSH
• Giardia lamblia genetika   izolace a purifikace   MeSH
• giardiáza epidemiologie   parazitologie   veterinární   MeSH
• kryptosporidióza epidemiologie   parazitologie   MeSH
• mikrosporidióza epidemiologie   parazitologie   veterinární   MeSH
• molekulární epidemiologie MeSH
• nemoci skotu epidemiologie   MeSH
• novorozená zvířata MeSH
• prevalence MeSH
• průřezové studie MeSH
• skot MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Čína epidemiologie   MeSH

A 23-year-old, HIV-negative woman who had undergone a hematopoietic stem cell transplantation was admitted to the hospital with respiratory failure and symptoms of bronchiolitis obliterans. A chest computed tomography scan revealed diffuse ground-glass opacification and fibrous plugs. Due to worsening respiratory failure despite treatment, ventilation was provided through a tracheostomy tube. Molecular examination of bronchoalveolar lavage and urine revealed Enterocytozoon bieneusi infection. After treatment with albendazole the patient gradually improved, but the pathogen was not eradicated and reappeared on follow-up examination. E. bieneusi belongs to the most clinically important microsporidial species infecting humans, mostly those who are immunocompromised. This fungus tends to infect enterocytes of the intestine, and there are limited studies concerning its extraintestinal location. This is the first report of a case of disseminated respiratory and urinary E. bieneusi infection in a transplant recipient.

• Klíčová slova
• Enterocytozoon bieneusi, Hematopoietic stem cell transplant, Respiratory tract infection,
• MeSH
• albendazol terapeutické užití   MeSH
• bronchoalveolární laváž MeSH
• Enterocytozoon účinky léků   izolace a purifikace   MeSH
• HIV infekce MeSH
• imunokompromitovaný pacient účinky léků   MeSH
• infekce močového ústrojí diagnóza   farmakoterapie   MeSH
• levofloxacin terapeutické užití   MeSH
• lidé MeSH
• mikrosporidióza diagnóza   farmakoterapie   MeSH
• mladý dospělý MeSH
• rizikové faktory MeSH
• transplantace hematopoetických kmenových buněk * MeSH
• Check Tag
• lidé MeSH
• mladý dospělý MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• Názvy látek
• albendazol MeSH
• levofloxacin MeSH

Microsporidia are obligate intracellular parasitic protozoa infecting the wide variety of hosts and are commonly known as a cause of chronic diarrhea particularly in immunocompromised individuals. Molecular-based tests have high sensitivity and specificity in disease diagnosis. However, these tests' performance relies on the isolation of DNA in a good concentration. The standard procedures of commercial DNA extraction kits are usually insufficient for this purpose due to the tough walls of spores. This study aimed to test the significance of pretreatments by glass beads and freeze-thawing processes in DNA isolation from microsporidia spores. The parasite was cultured in growing Vero cells and seven serial dilutions were prepared from the collected spores. DNA purification was performed according to different tissue kits and stool kit procedures with and without any pretreatment. Concentration of isolated DNA samples were evaluated by real-time PCR. As a result of this study, the detectable amount of spores is minimum 10 spores in each 100 μ! sample according to the different tissue kits' standard protocols. However, according to the DNA stool mini kit, the detectable amount of spores was found to be 1,000 spores/100 μl of stool sample when pretreated with both the freeze-thawing and glass beads methods.In conclusion, the current study demonstrated that further pretreatments are an essential process for DNA extraction from the stool specimens in order to avoid possible false negativity in the diagnosis of microsporidiosis.

• Klíčová slova
• DNA extraction, Microsporidia, freeze-thawing, glass beads, pretreatment,
• MeSH
• Cercopithecus aethiops MeSH
• DNA fungální izolace a purifikace   MeSH
• Enterocytozoon genetika   MeSH
• feces parazitologie   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• Microsporidia genetika   MeSH
• mikrosporidióza diagnóza   mikrobiologie   MeSH
• molekulární biologie přístrojové vybavení   metody   MeSH
• reagenční diagnostické soupravy * MeSH
• senzitivita a specificita MeSH
• spory hub izolace a purifikace   MeSH
• Vero buňky MeSH
• zmrazování MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• Názvy látek
• DNA fungální MeSH
• reagenční diagnostické soupravy * MeSH