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Diverzita kryptosporidií volně žijících psovitých a medvědovitých šelem
KELLNEROVÁ, Klára
The study was focused on study of diversity of Cryptosporidium spp. in wild canines and bears in the Czech Republic, Slovak Republic, Poland and Romania. A total of 359 faecal samples were collected from 179 red foxes (Vulpes vulpes), 83 grey wolves (Canis lupis), 63 brown bears (Ursus arctos) and 34 jackals (Canis aureus). Faecal samples were screened for Cryptosporidium by microscopy and PCR/sequencing. Phylogenetic analysis of small-subunit rRNA, actin and 60-kDa glycoprotein sequences revealed the presence of C. tyzzeri, C. andersoni in red foxes, C. canis and C. ubiquitum in gray wolves and C. galli in a brown bear and a red fox. Subtyping of C. ubiquitum and C. tyzzeri isolate by sequence analysis of the 60-kDa glycoprotein gene showed that isolates belonged to the XIId and IXa subtype family, respectively. Detection of host-non-specific cryptosporidia, except C. canic and C. ubiquitum, in wild canine and bears shows rather a food preference of screened carnivors than on an active infection.
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Losses of mitochondria and plastids in the evolution of eukaryotes
Trokšiar, David ; Hampl, Vladimír (advisor) ; Hadariová, Lucia (referee)
- 5 - Abstract: Mitochondria and plastids were acquired by endosymbiotic event, where prokaryotic organism was engulfed by ancestors of extant eukaryotes. There are more known endosymbiotic events in plastid evolution. In primary endosymbiosis cyanobacterium cell was engulfed by heterotrophic eukaryotic organism. In following secondary, tertiary and quaternary endosymbiotic events eukaryotic cell was engulfed by another eukaryote. Mitochondria originated by engulfment of α-proteobacteria. In the evolution of eukaryotes, reduction of mitochondria occurred in many lineages, making living under anaerobic conditions possible. The least reduced form is anaerobic mitochondria, which together with aerobic mitochondria and hydrogen producing mitochondria, possess genome. Hydrogenosomes and the most reduced form mitosomes, does not possess genome. Plastid reductions led to loss of photosynthetic ability. In last years, more examples of organisms that lost entirely their semi- autonomous organelle, are coming. Loss occur at two parasitic representatives of the Alveolata group, and one endobiotic oxymonad. Parasites Cryptosporidium parvum and Hematodinium lost nonphotosyntetic plastid, whereas Monocercomoides lost its mitochondria. Semi-autonomous organelles were dispensable, because all representatives have access to...
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Presence of specific DNA and coproantigen of Cryptosporidium as an indicator of ongoing infection
TOMANOVÁ, Vendula
Cryptosporidium is a genus of apicomplexan unicellular epicellular parasite with worldwide distribution causing watery diarrhea in humans and animals. The life cycle is completed in one host, where Cryptosporidium parasitizing epithelial cells of gastrointestinal tract and in birds can cause disease of respiratory or urogenital tract. Course of disease depends on condition of immune system. For immunodeficient individuals could be life threatening. One of problems especially in developing countries is early and correct diagnostic, particularly no effective treatment currently exist. The aim of this thesis was to compare efficiency of immunochromatographic tests in samples stored under different conditions. The comparison of sensitivity and specificity of these tests with molecular and microscopic techniques was also performed. Additionaly, suitability of immunochromatographic tests for detection of active infection during prepatent period was evaluated. The theoretic part includes general information about Cryptosporidium. Its taxonomy, cycle of evolution or transmission and course of disease. Using of immunochromatographic test is also mentioned. No differences in sensitivity of used immunochromatographic tests was observed in this thesis. The detection rate for most of tests was 200 oocyst per sample. The presence of coproantigen is depend upon presence of oocysts in a sample. False negative results of immunochromatographic assays was caused by i) low concentration of oocysts in a sample (sensitivity) or ii) antibodies in used test don´t react with antigen of Cryptosporidium spp. (specificity). Results of this thesis show that combination of immunochromatographic tests and other techniques is convenient. During prepatent period is not possible to detect specific DNA, antigen or oocysts of Cryptosporidium. The active infection could not be distinguish from passage of oocysts using of immunochromatographic assays even if PCR is also used.
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Card of Cryptosporidium infections by humans and animals
BLÁHOVEC, Ondřej
The representatives of the Cryptosporidium genus are one of the causes of gastrointestinal tract diseases in humans and animals. In animals the host may even be a respiratory apparatus. The disease is called cryptosproridiosis. In majority of cases this infection can bypass without any major problems, but in immunosuppressed individuals it can cause serious health problems. Cryptosporidium has a monoxenous development cycle, which basically means that the entire development takes place in a single host. Exogenous stage is represented by oocysts, which are in case of a gastrointestinal disease excreted in faeces. In case of a respiratory disease the oocysts make they way out via respiratory and nasal secretions. This leads to contamination of the environment or water. In general, it is expected that Cryptosporidium isolates, which are present in one class of vertebrates, are not infectious to a non-specific host from other classes. It is also expected that cryptosporidia have low host specificity. But this does not exclude that some kinds have gradually extended its specificity to more species. It is also apparent that cryptosporidium infections are common in animals that inhabit the external environment, so even a human can be endangered by this zoonosis, although the incidence in the Czech Republic is low. The reason for the low numbers may as well be that parasitological examination is not performed very often, so the estimated prevalence in the population is probably much higher. Therefore to reveal the originator of this disease it would be appropriate to perform a parasitological examination in persons who are in contact with animals, this way the cryptosporidium infection would be excluded or proven.
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Cryptosporidium Diversity of Selected Bat Species (Chiroptera)
HOŘICKÁ, Anna
Total 262 samples of different Chiropterans originating from 27 localities in the Czech Republic were collected. Occurrence and prevalence of Cryptosporidium infection in bats were screened using staining method and molecular tools. While no Cryptosporidium oocyst was detected by microscopy, PCR analyses revealed presence of three positive specimens from three common pipistrelle (Pipistrellus pipistrellus). Zoonotic C. parvum was detected in one case and novel Cryptosporidium bat genotype III was found in other two samples.
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Prevalence and diversity of Cryptosporidium infecting fur animals
KELLNEROVÁ, Klára
The object of this thesis was evaluation of occurrence and prevalence of Cryptosporidium infection in fur animals, mainly American mink, fox and chinchilla. A total 370 individual specimens originated from mink (n = 340), fox (n = 18) and chinchilla (n = 12) were collected. While microscopy examination did not proved any presence of Cryptosporidium oocyst in fecal samples, molecular tolls based on amplification of small ribosomal subunit and 60 kDa glycoprotein of Cryptosporidium revealed three positive samples in minks. Following phylogeny analyses of both loci showed presence of C. ubiquitum of the XIIa1 family subtype in all positive samples. The XIIa1 family subtype was detected in Carnivores for the first time. No correlation between Cryptosporidium infection and presence of diarrhea was observed in this thesis.
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Biology and diversity of Cryptosporidium infecting squirrels (Sciuridae)
BARVÍŘ, Pavel
In total 399 samples from three species of squirrels (Sciurus vulgaris, Sciurus carolinensis and Callosciurus finlaysonii) were collected from 2010 to 2013. All samples were examined for the presence of Cryptosporidium using by molecular methods, which demonstrated by presence of Cryptosporidium specific DNA in 18 samples (4.5%). Out of them 12 samples were positive onCryptosporidium ferret genotype, 3 on Cryptosporidium skunk genotype, 1 on Cryptosporidium chipmunk genotype I and 2 samples onCryptosporidium ubiquitum. Statistical analyses did not reveal any influence of gender on the occurrence of Cryptosporidium. Juvenile individuals of squirrels from the family Sciuridae are more often infected by Cryptosporidium than the adult animals.
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Cryptosporidial infection in hedgehog
HUCLOVÁ, Kristýna
This study involves the morphological, biological, and molecular characteristics of Cryptosporidium spp. in hedgehogs. Course of infection based on 405 isolates from 15 hedgehogs obtained during a few months was observed. Morfological and molecular analyses were conducted to detect oocysts of Cryptosporidium spp. Altogether 69 (17.01%) samples from 11 hedgehogs were morfologically positive and 81 (19.9%) samples were molecular positive. Only 4 individuals remained negative, the cumulative prevalence represented 73.3 % of total samples. Oocysts from monitored hedgehogs measuring 4.94,7 m (mean = 4.8 m) × 4.0-3.8 m (mean = 3.9 m) with a length to width ratio of 1.22 (1.261.20) (n = 50) in native were indistinguishable from those of C. erinacei and C. parvum. Molecular analyses based on small subunit rRNA, 60 kDa glycoprotein and Cryptosporidium oocyst wall protein revealed presence of C. parvum and C. erinacei. Cryptosporidium parvum IIdA18G1 was detected in 11 hedgehogs. Mixed infection with C. parvum with C. erinacei was observed in 3 animals. The C. parvum IIdA18G1 genotype has never been described in hedgehog before. It belongs to zoonotic IId subtype family frequently found in goats and lambs. This subtype was identified in lambs from Great Britain and Spain and also in children from Kuwait. Cryptosporidium erinacei is adapted to hedgehogs and it does not appear to cause clinical disease in hedgehogs. All positive individuals were juvenile and wild hedgehogs.
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Diversity of Cryptosporidium spp. infecting rodents from subfamily Arvicolinae in the Czech Republic
HÁJKOVÁ, Ivana
Abstract In order to examine the prevalence of Cryptosporidium in wild Arvicolinae in the Czech Republic and understand the role that wild rodents play in the transmission of this parasite to humans and livestock, 152 faecal samples from 129 common voles (Microtus arvalis) and 23 bank voles (Clethrionomys glareolus) were collected on 9 localities in 2012. All samples were examined for presence of Cryptosporidium sp. using both the aniline-carbol-methyl violet staining method and molecular tools. The age, sex and faecal consistency were noted at the time of sampling. Microscopical examination revealed the presence Cryptosporidium sp. oocysts in 2 samples originated from common voles and 2 samples from bank voles. Genotyping was done through PCR amplification and characterization of the SSU rRNA and actin loci. Cryptosporidium specific DNA was detected in 10 samples (4 from common voles and 6 from bank voles) including those microscopically positive. Cryptosporidium infection was not linked to diarrhoea. Sequence and following phylogeny analyses revealed two new Cryptosporidium genotypes originated from bank voles and two new genotype from common vole, phylogeneticaly distinct from known species and genotypes. The host specificity needs to be verified by experimental infection in the future.
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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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