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The incidence of potential agents of parasitic zoonoses in Svalbard
HONSOVÁ, Lucie
In my thesis I have studied the occurrence of selected parasitic zoonoses in terrestrial vertebrates on Svalbard. Parasitic examination was focused on ascertaining of the presence of cryptosporidia, microsporidia and giardia. There were 87 samples of excrements of 12 animal species. In Svalbard, only 3 species of terrestrial mammals occure. Originally there were only the svalbard reindeer (Rangifer tarandus platyrhynchus) and the arctic fox (Alopex lagopus). In the first half of the 20th century east european vole (Microtus levis) has been introduced there probably from Russia ? region of Petersburg. In this archipelago there can be found different birds species in summer and the only one who winter there is the rock ptarmigan (Lagopus muta). The samples were analyzed by microscopy of stained smears and by molecular methods. For microscopic examination the staining by Miláček ? Vítovec was used to find out the presence of oocysts of cryptosporidia in animal excrements. The molecular diagnostics of cryptosporidia, microsporidia and giardia was done using the polymerase chain reaction (PCR). The positive detection was followed by the sequential analyses which proved the presence of cryptosporidia: C. parvum in Anser brachyrhynchus, Cryptosporidium goose genotype II in Branta leucopsis and C. muris TS 03 in Rangifer tarandus platyrhynchus. Also the presence of Encephalitozoon. cuniculi genoype II in three samples of Rangifer tarandus platyrhynchus, one sample of Alopex lagopus and one sample of Branta leucopsis was proved. Another recognized kind of microsporidia was Enterocytozoon bieneusi in Rangifer tarandus platyrhynchus and Anser brachyrhynchus. Both findings present new genotypes of microsporidia. These findings proved that the extreme conditions of high Arctic on Svalbard enable spreading of intestinal unicellular parasites, cryptosporidia and microsoridia.
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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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Detection of cryptosporidia by means of molecular methods in clinical samples:infection or the transit of oocyst through the host gastrointestinal tract?
MUNZAROVÁ, Lucie
Representatives of the Cryptosporidium family are, from a medical and veterinary aspect, designated as important unicellular parasitic protozoa infecting all classes of vertebrates including humans. Their development cycle is monoxene, which means that its life cycle is completed uniquely in one host. Their exogenous stages of infection development ? oocysts ? are mostly secreted in faeces causing a parasitic disease called Cryptosporidiosis. Generally, it is assumed that isolated cryptosporidia from one class of vertebrates are not contagious for other hosts from different classes, and also, that the majority of cryptosporidia types and genotypes have low host specifity. However, there are many cases where the presence of oocysts was proved, or the presence of specific cryptosporidium DNA in faeces of atypical hosts. With regard to the fact that the majority of genotypes and their potential hosts were described only on the basis of the presence of the specific cryptosporidium DNA in faeces, the question arises as to whether the aforementioned hosts are in all cases perceptive to the given infection, or it was only contamination in a sample or the transit of oocysts through the host gastrointestinal tract. This issue is also characterised as being the main objective of this thesis. The study gives an appropriate answer to this question by a series of experiments. Different isolations of cryptosporidia were used for experimental infection of rodents which were per orally infected and put down after reaching the specified patent period. By means of the combination of standard parasitological and molecular methods, the presence of oocysts and specific DNA of cryptosporidia in faeces and in mucous membrane of the gastrointestinal tract of infected hosts was observed. The results of this study prove that the transit of oocysts through the gastrointestinal tract can be detected in the interval between 6 and 24 hours after infection, yet not in every case. On the basis of this finding, the first hypothesis which stated that it is not possible to qualify explicitly by detection of a specific cryptosporidium DNA in faeces whether the infection has begun in the host or it is the transit of oocysts through the host gastrointestinal tract, was thus partially proved. On the other hand the second hypothesis stating that the specific cryptosporidium DNA in clinical samples of faeces can be detected only in cases of the host´s successful infection, was rejected.
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The occurrence of Coccidiosis in the intestine of sucking pigs before and after weaning
KOTILOVÁ, Jiřina
In two years of observation, (spring 2006, autumn 2006, spring 2007 and autumn 2007) were being screened for parasites in total 495 faecal samples coming out of three farms from Ceske Budejovice (285 samples of sucking pigs not older then 28 days and 174 samples of piglets not older then 8 weeks). The method used to examine those samples was a flotation-concentrating method (Sheather{\crq}s carbohydrate fusion) and in the year of 2007 was also used a specific aniline-carbol-methyl violet staining method to detect the Cryptosporidium spp. followed by positive molecular characterized (direct sequencing of partial SSU rRNA partial genes and PCR-RFLP at the SSU rRNA). In screened samples were mainly detected parasites named Cryptosporidium spp., found in 4,1% of faecal samples in 2006 and in 2007 in 32,8% faecal samples, out of which 14,4% was found in pre-weaned piglets samples and 26,4% in post-weaned piglets. Based on genotyping provided on positive samples out of the year 2007, using method of sequensing analysis SSU rRNA, was the occurence of C. suis, Cryptosporidium pig genotype II aC. Muris described. High prevalence of Isosporou suis 13,9 % (64/459) was also detected with its appearance, in particular, in pre-weaned piglets 21,4 % (61/285). Further on, some of other identified group was Eimerie spp. 5,7 % (26/459) infecting, in the main, post-weaned piglets 10,9% (19/174) and Giardia intestinalis 2,4 % (10/459). Most of the samples mentioned occured in conditioned faeces and there is no seasonal relationship to the parasital occurance.
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Cryptosporidial infections of pigs
FORNBAUMOVÁ, Iva
Recent genetic and biological characterization studies have identified 2 distinct apparently host-adapted genotypes of Cryptosporidium in pigs, that is, Cryptosporidium suis and the Cryptosporidium pig genotype II. The infection of both the above mentioned Cryptosporidium appears to occur a much less severe disease in pigs than it does Cryptosporidium parvum. Sporadic cases of C. suis in human have been reported, but the risk of infection for human is uncertain.
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Cryptosporidial and microsporidial infection on the pig farms, in the Czech Republic
HANZLÍKOVÁ, Dagmar
The cryptosporidia and microsporidia are worldwide spread opportunistic parasites. With regard to their wide host spectra and zoonotic potential there is a significant risk for immunocompromised humans, especially AIDS patients. Natural Cryptosporidium infections in pigs are widespread but generally apathogenic. A total of 411 faecal samples, 39 of sows, 127 of pre-weaned, 122 of post-weaned piglets and 123 of pre-growers from three farms were screened for Cryptosporidium spp. infection using the aniline-carbol-methyl violet staining method and positive molecular characterized (direct sequencing of partial SSU rRNA partial genes and PCR-RFLP at the SSU rRNA). Overall, infections were detected in 84 out of 411 animals (20 %), with highest infection rates among post-weaners (27 %). All positive samples were genotyped based on SSU rRNA sequence analysis. C. suis, Cryptosporidium pig genotype II, and C. muris were detected. The microsporidial infections are very frequent in breeds of pigs. Pigs have been frequently reported to be infected with E. bieneusi. Spores were detected in 258 samples of 342 (75 %). The high prevalence was determined in pre-growers (86 %). Genotyping based on the ITS regions of the rRNA gene identified that most pigs were infected with the species-specific genotype F, while two animals had the zoonotic genotype D and two had genotype Peru 9.
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The role of rotifers by filtration of \kur{Cryptosporidium} in water column
BRŮČKOVÁ, Petra
Filtration abilities of Cryptosporidium parvum oocysts by rotifers (Brachionus calyciflorus) were studied. The filtration rate of rotifers is quick: the passage of oocysts through the digestive system is few minutes only. During the first 10 minutes 24300 of whole amount one million oocysts passed through the digestive system of 100 rotifers. Moreover, 93% of oocysts were death after passage. All oocysts expelled were unable to infect suckling BALB/c mice. Oocysts were degraded in mastax, stomach and intestine. Rotifers can be use for detection of cryptosporidia oocysts in water and they effectively decrease their numbers.
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