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Poultry cryptosporidiosis
KURAL, Vladimír
A total of 270 samples of domestic hen (Gallus gallus f. domestica) from 20 farms were collected during two consecutive years (from 2011 to 2012). Microscopical examination of aniline-carbol-methyl violet stained fecal smears revealed 5 positive samples originating from one farm. 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 8 samples as a novel genotype, working titled Cryptosporidium hen genotype. The sequences identified as hen genotype matched most closely with Cryptosporidium bovis which was originally reported from cattle in the most cases.
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Cryptic diversity of free-living trichomonads and their phylogenetic position within Parabasalia
Céza, Vít ; Čepička, Ivan (advisor) ; Hampl, Vladimír (referee)
Trichomonads (Parabasalia) are anaerobic microeukaryotes classified in the supergroup Excavata. Inclusion of parabasalids within Excavata is exclusively based on the molecular- phylogenetic evidence. Over 400 species of parabasalids have been described so far, and the vast majority of them are endobiotic. In contrast, only few species of free-living parabasalids forming four independent lineages have been described (Pseudotrichomonas keilini, Ditrichomonas honigbergii, Monotrichomonas carabina, Honigbergiella sp., Tetratrichomonas undula, and Lacusteria cypriaca). Lacusteria cypriaca is a new species and genus described in our recent paper. In this paper we published the first two sequences of SSU rDNA from Pseudotrichomonas keilini as well. All of these lineages are likely secondarily free-living, and they developed from endobiotic ancestors. In addition to the already published Lacusteria cypriaca and Pseudotrichomonas keilini strains, we have recently obtained seven another isolates of free-living trichomonads (LAGOS2D, E2NT, CK, LAGOS2M, GR8, GOU23 LIVADIAN, and VAV1A1); from all of these isolates we sequenced SSU rDNA and performed phylogenetic analyses. These isolates split into four independent evolutionary lineages, which indicate that free-living parabasalids are more diversed and...
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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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Variabilita polymorfizmu v genu FABP3 a jeho asociace s ukazateli kvality masa u prasat
Boleslavová, Štěpánka
The aim of the study was to study the variability of the gene in the population FABP3 pig breed Czech large white. Analyze their association with indicators of pork production. Gene FABP3 belonging to multigene families FABP (fatty acid binding proteins affecting the transport of long chain fatty acids). Analyzed polymorphism was at position X98558:g.1321G>C in the 5'UTR region. The relative frequencies of genotypes were: HH = 0,87, Hh = 0,13. hh genotype was not detected in the monitored individuals. The values of the relative frequencies of alleles were as follows: H = 0,93 h = 0,07. Based on the association analysis showed a statistically significant difference between genotypes for stearic acid and linoleic acid. Highly statistically significant difference was observed with palmitic acid.
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Studium SNP genu DGAT1 jako kandidátního genu pro kvalitu vepřového masa
Tomášková, Marie
This work was focused on the study of variability DGAT1 gene in the pig population Czech Large White breed. Subsequently, association analysis was performed of the gene and the different production indicators of quality of pork meat. Examined polymorphism was found at position 103 of intron 2 of chromosome 4. DGAT1 gene has a major role in the synthesis tryacylglycerols and may affect the storage of fat in the body. Relative genotype frequencies were: AA = 0.4222; AG = 0.4889; GG = 0.0889. The values of the relative frequencies of alleles were as follows: A = 0.6666 and G = 0.3334. The association analysis didn't show any statistically significant differences.
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Mutations in MLH1 gene and MSI status as molecular characteristics of sporadic colorectal cancer
Čaja, Fabián ; Vodička, Pavel (advisor) ; Kadlecová, Jitka (referee)
Colorectal carcinoma (CRC) is one of the most prevalent malignancies in the Czech Republic. In general, there are two molecular pathways leading to CRC: one is characterized by chromosomal instability, the other by the deficiency in DNA mismatch repair (MMR) genes. MutL homologue 1 (MLH1) gene, a member of the MMR gene-family, represents a key component of the MMR system, responsible for recognition of nucleotide mismatches occurring during DNA replication, and for the recruitment of repair proteins to correct the replication errors. According to literature, somatic mutations in MMR genes, and MLH1 in particular, hallmark sporadic, MMR deficient, CRC cases. We aimed at analyzing somatic events in MLH1 gene and the determination of microsatellite instability (MSI) status in 99 DNA samples from 96 patients with sporadic CRC. Mutations were screened by high resolution melting (HRM) curve analysis. Positive cases in each run were subsequently verified by automated sequencing. Mainly gene variants were found in MLH1 gene: We discovered two new variants, one in exon 2 at position c. 204 C>G, p. Ile68Met (98 C/C, 1C/G) and the other in exon 11 at position c. 973 C>T, p. Arg325Trp (98 C/C, 1 C/T). Only the latter variant c. 973 C>T was identified as somatic mutation. All other variants found in MLH1 gene...
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Larvae of flesh fly Neobellieria bullata as a source for novel antimicrobial peptides
Neubauerová, Tereza ; Macková, Martina ; Macek, Tomáš ; Šanda, Miloslav ; Voburka, Zdeněk
The antibiotic resistant strains of microorganisms are increasing threat for public health. Antimicrobial peptides from natural sources display a wide spectrum of activity. In our laboratory, several peptide fractions with antimicrobial acivity against gram-positive and gramnegative bacteria and fungi were isolated from larvae of flesh fly Neobellieria bullata. The presence of peptides was proved by tricine electrophoresis, molecular weights and sequences determination by MS and Edman sequencing, respectively. In comparison with database sequences we obtained structures present mainly in insect and amphibian antimicrobial peptides, but total matches were not higher than in 36%. Upon the obtained sequence motifs we designed structures of several synthetic peptides. These were synthetised on the Rink amide MBHA resin. Synthesis was performed according Fmoc/PyBOP-DIEA protocol. Antimicrobial activity of these peptides was determined.
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Design of stable antimicrobial peptides through hydrocarbon stapling
Chapuis, Hubert Jean ; Slaninová, Jiřina ; Monincová, Lenka ; Bednárová, Lucie ; Čeřovský, Václav
From the venom of wild bee Lasioglossum laticeps we have recently isolated novel antimicrobial peptides named lasioglossins. One of them, LL-III (VNWKKILGKIIKVVK-NH2), is an amphipathic α-helical peptide which shows strong antimicrobial properties and a low hemolytic activity. We anticipated that the incorporation of an all-hydrocarbon staple (bridge) into the LL-III sequence could increase its propensity to form an α-helix and lead to an improvement of its proteolytic stability as well as increase the antimicrobial activity. LL-III analogs featuring olefinic side chains in various positions were prepared by solid phase peptide synthesis. Ring closing olefin metatheses catalyzed by Grubbs-I catalyst were carried out on the solid support, either between i and i+4 positions or between i and i+7 positions.
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Control Techniques of Grid Connected PWM Rectifiers under Unbalanced Input Voltage Conditions
Bejvl, Martin ; Šimek, Petr ; Škramlík, Jiří ; Valouch, Viktor
Current-controlled voltage source converters are widely used in grid-connected applications,for example at ac drives with indirect frequency converters.The structure and parameters of the PLL are developed and proposed in order to cope with the grid containing both the positive and the negative sequence component, and minimize the wrong frequency transients during phase angle steps and also in the start-up stage. The DSC technique was realised. There is also necessary to compensate the negative sequence component in the grid voltage. The negative sequence component of the grid voltage causes ripple of the dc voltage in the intermediate circuit. Several sophisticated topologies of converter current controller were developed, simulated and tested for this purpose. Results of simulation and experimental tests are provided to evaluate different current control schemes and phase locked loop techniques.
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