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Faktory ovlivňující excystaci žaludečních kryptosporidií hlodavců
SCHULZOVÁ, Tereza
The aim of this study was to determine the effect of host body temperature and en-vironmental pH on the excystation and preservation of the ability to infection of rodent gastric cryptosporidia oocysts. The infectivity of unexcysted oocysts was further examined. Immunodeficiency SCID mice (Mus musculus) were infected with C. proliferans oocysts, then their faeces were collected and individual experi-ments were performed. The effect of temperature on excystation, when oocysts were placed in water baths at the appropriate temperature (32-44 °C) and the effect of pH, when oocysts were exposed to solutions of different pH (2-7), was investi-gated. Cell separation was then performed using a flow cytometer and the values obtained were recorded in the tables. The most oocysts excysted at 40 °C and pH 7. An autopsy of infected mice was also performed to determine the infectivity of oo-cysts from various parts of the digestive system. All taken oocyst samples were infectious for the subsequent host. Infection experiments, which were further car-ried out revealed, that unexcysted oocysts exposed to hypochlorite and laboratory temperatures retain their ability to infect. In contrast, the oocyst sporozoites that excysted were susceptible to both hypochlorite and room temperature, thus losing the ability infect the subsequent host.
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Diverzita endoparazitů lišek polárních
PAVLOVIČOVÁ, Markéta
The foxes represent a reservoir of zoonotic infections therefore it is important to be familiar with an occurrence of possible diseases. Arctic fox (Vulpes lagopus) faeces were collected between 2003 and 2011 from various locations on Bylot Island, Canada. These samples were consequently treated with microscopic and molecular methods to prove the presence of microsporidia, cryptosporidia and giardia. The presence of a specific DNA of Cryptosporidium was detected in two samples and specific DNA of Microsporidia was detected in 34 samples for genus Encephalitozoon and in five samples for genus Enterocytozoon. None of the animals showed any symptoms of a diarrhoea. Subsequent analyses showed the presence of the Cryptosporidium canis fox genotype, Encephalitozoon cuniculi II and III genotypes and Encephalitozoon hellem genotype 1A. None of the examined faeces samples were positive for the presence of specific G. intestinalis DNA. Results of the thesis show that Artic foxes are mainly infected by microsporidia of the genus Encephalitozoon, while the occurrence of other observed parasites was rare.
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OCCURRENCE AND GENOTYPISATION OF CRYPTOSPORIDIES IN OSTRICHES IN FARM BREED
HEJZLAROVÁ, Adéla
Avian cryptosporidiosis is a common parasitic disease caused by unicellular parasites belonging to the genus Cryptosporidium spp. These parasites have been described as a cause of various respiratory and gastrointestinal diseases of humans, farm and wild animals. Total of 204 faecal samples of common ostriches (Struthio camelus) were collected on four ostrich farms in the Czech Republic. Samples were screened for Cryptosporidium presence by microscopy and PCR/sequencing. Overall, 2.5 % (5/204) and 5.9 % (12/204) of animals were positive for Cryptosporidium by microscopy and PCR, respectively. Phylogenetic analysis of small subunit rRNA, actin and gp60 gene sequences showed the presence of Cryptosporidium avian genotype II (n=7) and C. ubiquitum 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 6.13 × 5.15 m, which is indistinguishable from C. baileyi and C. avium. Cryptosporidium avian genotype II was experimentally infectious for geese (Anser anser f. domestica), chickens (Gallus gallus f. domestica) and cockatiels (Nymphicus hollandicus), with a prepatent period of four, seven and eight days post infection, respectively and not infectious for mice (Mus musculus) and rats (Rattus norvegicus). The infection intensity ranged from 1,000 to 16,000 oocysts per gram. None of the naturally or experimentally infected birds developed clinical signs in the present study.
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Kryptosporidiové infekce veverek
ŠVAJLENOVÁ, Kamila
We investigated the Cryptosporidium spp. infecting red squirrels (Sciurus vulgaris) in 27 areas in the Czech Republic and Slovakia. Examination of 157 faecal samples revealed oocysts of Cryptosporidium sp. in 14 animals (21.98%). Phylogenetic analysis of small subunit rRNA, actin and gp60 gene sequences showed the presence of Cryptosporidium infection in 14 animals. Sequence analysis revealed the presence of Cryptosporidium ferret genotype in all positive samples. Three gp60 subtypes, VIIIb, VIIIc and novel subtype VIIIe were detected within screed population of squirrels. The occurrence of Cryptosporidium infection did not differ between the age or sex of animals. Only juvenile animals shed microscopically detectable amount of oocysts. The infection intensity ranged from 100000 to 250000 oocysts per gram. None of the naturally infected squirrels developed clinical signs in the present study. Cryptosporidium chipmunk genotype I obtained from naturally infected Eastern gray squirrel (Sciurus carolinensis) in Italy and used for experiments was infectious for mice (Mus musculus; strains SCID, C57BL/6J, CD4-/- and CD8-/-), ferret (Mustela putorius furo) and red squirrels, with a prepatent period of 10, 4, 4, 4, 4 and 11 days post- infection, respectively. Oocysts of the Cryptosporidium chipmunk genotype I measured 5,64 (5,50-5,89) × 5,37 (4,86-5,60) m (shape index 1,05 (1,01-1,14)). All mice strains except SCID clear infection within 8-18 days post-infection. Clinical cryptosporidiosis was observed in SCID mice, ferret and squirrels, but only squirrels suffer severe diarrhoea and the infection was lethal for them. SCID mice represent the suitable laboratory model host for maintenance of Cryptosporidium chipmunk genotype I.
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Biologie a diverzita kryptosporidií infikujících myši domácí (Mus musculus)
PRANTLOVÁ, Veronika
Cryptosporidium spp. are globally distributed unicellular parasitic protozoa of the phylum Apicomplexa that infect a wide range of vertebrates, including humans. Their exogenous developmental stages are resistant to most disinfectants and no effective drugs have been developed to date. Cryptosporidium infections of humans and many livestock have been well studied over the past 30 years, but our knowledge of Cryptosporidium spp. in small mammals has been inadequate. Recently, detailed studies of cryptosporidia have been carried out in a number of rodents, especially mice, voles, rats or tree and ground squirrels. This thesis aims to add to our mosaic of knowledge data on the prevalence, diversity and biological characteristics of cryptosporidia in the house mouse (Mus musculus), the most common unwelcome visitor to human settlements.
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Kryptosporidiové infekce ryb
HAVELKA, Michal
Cryptosporidium spp. is able to infect a wide range of hosts including wild and domestic animals, and humans. Although it is well known that fish could be host several Cryptosporidium species and genotypes, little information is available concerning the prevalence of Cryptosporidium in fresh water fish in Europe. Additionally, the infectivity of C. parvum for fish, the species with wide host range, remains poorly studied. In this study we collected 283 specimens of wild fresh water fish from eight localities, 41 specimens of ornamental fish collected from eight pet shops in the Czech Republic. The occurrence and genetic diversity of Cryptosporidium spp. in studied fish was examined by microscopy and polymerase chain reaction (PCR)/sequencing targeting the small subunit rRNA. The infectivity of C. parvum for pike (Esox lucius) and crucian (Carrasius auratus) were experimentally verified. Microscopy and molecular analysis did not revealed presence of oocysts and specific Cryptosporidium DNA in wild fresh water fish. While microscopy analyses of faecal samples of ornamental fish were negative for presence of Cryptosporidium spp. oocysts, molecular analyses shown presence Cryptosporidium avium, Cryptosporidium baileyi, Cryptosporidium muris, Cryptosporidium parvum, Cryptosporidium tyzzeri and Cryptosporidium varanii. Histology did not show the presence of any developmental stages in any pikes and crucians inoculate with C. parvum or inhabit an environment with C. parvum oocysts. The presence of C. parvum DNA was detected only in intestinal contains of pikes and crucians who inhabit an environment with C. parvum oocysts. Presence of C. avium, C. baileyi, C. muris, C. parvum, C. tyzzeri and C. varanii in ornamental fish is probably resulting of the contamination of the water in the tank by caretaker and detected species almost certainly originated from the pet rodents and birds bred in the same shop. The result of the experimental study supports the hypothesis that cryptosporidia of mammals, birds and reptiles are not infectious for fish and their presence in the fish is result of the environmental contamination.
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Susceptibility of pigs to various Cryptosporidium species and genotypes
KESTŘÁNOVÁ, Michaela
Three-four and 8 week old pigs (three animals per isolate) were inoculated with Cryptosporidium muris C. tyzerri, C. suis and Cryptosporidium pig genotype II at a dose of 1 × 107 oocysts per animal. Pigs inoculated with C. muris or C. tyzerri showed no detectable infection and no clinical symptoms of cryptosporidiosis during 30 days post infection (DPI), and no macroscopic changes were detected in the digestive tract after necropsy. Any developmental stages were detected in gastrointestinal tract tissue neither by histology nor PCR throughout the duration of the experiment. The infectivity of these isolates was verified on SCID mice, in which oocysts shedding started from 4 to 8 DPI. Experimental infection revealed susceptibility of both 4 and 8 week old pigs to C. suis. While parasitological, molecular and histology examination confirmed susceptibility of 8 week old pigs to Cryptosporidium pig genotype II, 4 week old pigs were not being infected with this genotype. Both C. suis and Cryptosporidium pig genotype II were detected in small and large intestine. Based on our findings, it can be concluded that pigs are not susceptible to C. muris or C. tyzzeri infection, C. suis does not have age specificity and Cryptosporidium pig genotype II is not infectious for pre-weaned pigs
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Cryptosporidial infections of pets
MAROUŠOVÁ, Michaela
This thesis was focused on the incidence and prevalence of Cryptosporidium spp. in selected zoo boutiques and companion animals in České Budějovice. A total of 104 fecal samples of companion animals (Mus musculus, Rattus norvegicus, Cricetinae ssp., Cavia aperea porcellus, Oryctolagus cuniculus) were examined during 2011 to 2012. Samples were investigated by microscopic methods; positive samples were investigated by PCR. Cryptosporidium infections were observed only in mice (Mus musculus) and rats (Rattus norvegicus). There were detected two species of zoonotic cryptosporidia, C. tyzzeri and C. muris. The results show that animals bred in hobby breeding are infected by cryptosporidia and they should represent health risk for human.
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Endoparasitosis of feathered game
BARVÍŘ, Pavel
A total of 180 samples, 119 from pheasant (Phasianus colchicus) and 61mallard (Anas platyrhynchos) were collected during two consecutive years (from 2011 to 2012). All samples were examined for the presence endoparasite using the flotation method according Sheather. In samples from pheasants were detected these types of parasites: Capillaria spp., Eimeria spp., Heterakis spp., and Trichostrongylus spp. In addition, Syngamus spp. and Cryptosporidium spp. were detected in the duck samples. Microscopical examination of aniline-carbol-methyl violet stained fecal smears revealed to Cryptosporidium spp. 12 positive samples originating from Anas platyrhynchos. DNA was extracted from Cryptosporidium positive samples and all microscopically negative samples. Nested PCR was performed to amplify the partial SSU rRNA gene of Cryptosporidium. The sequence analyses of PCR-positive specimens identified 10 samples as Cryptosporidium avian genotype III and 4 samples as C. muris. No clinical signs were detected in any Cryptosporidium positive animals. This is the first report of Cryptosporidium avian genotype III in the Czech Republic and also in Anas platyrhynchos.
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