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Detection of antimicrobial resistance in sexually transmitted infections
KREJČÍKOVÁ, Pavlína
Sexually transmitted diseases (STDs) pose a public health risk, especially in terms of the rise of antibiotic resistance. This thesis summarizes the knowledge on these infections, most common causative agents, laboratory diagnosis and current epidemiology, antimicrobial resistance and its molecular mechanisms. Determination of antimicrobial susceptibility in most STD pathogens is not possible in routine microbiology due to high or specialized culture requirements. Also, because of the higher sensitivity of detection, molecular biology methods are used to diagnose STDs, which negates the need to perform culture in Neisseria gonorrhoeae. The experimental part of the project aimed to detect the described resistance mechanisms in positive samples tested by multiplex PCR for the presence of STD agents. In the first part, we used whole genome sequencing (WGS). Unfortunately, the sequences obtained contained a large number of human sequences and thus the extra-human sequences did not provide sufficient coverage. The same result was achieved after a sample pretreatment with saponin and Triton-X followed by purification. Due to the unsuitability of WGS, we attempted to optimize and implement previously described target-specific methods to detect the most common resistance mechanisms in Neisseria gonorrhoeae. The first method was PCR amplification with melting curve analysis (HRM). As a reference method to HRM, we used Sanger sequencing of six genes (penA, mtrR, ponA, porB, 23S rRNA, gyrA). During this work, we optimized the temperature profiles for PCR amplification of individual genes. To cover the analyzed DNA fragment (part of the gene) it was necessary to design additional sequencing primers. The clinical use of HRM for the detection of resistance mechanisms in N. gonorrhoeae needs to be validated on DNA samples with identified resistance mechanisms by specific fragment sequencing, which was optimized in the present work.
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Rezistence blýskáčka řepkového (Brassicogethes aeneus, Fabr. 1775) k vybraným účinným látkám insekticidů
Langová, Melanie
The aim of this thesis was to evaluate the status of resistance of the pollen beetle (Brassicogethes aeneus, Fabr. 1775) to selected active ingredients of insecticides in 2020. The tested active ingredients were lambda-cyhalothrin, tau-fluvalinate, indoxacarb and Biscaya 240 OD (thiacloprid) according to IRAC testing methodology (Insecticide Resistance Action Committee) 011, 021 and 027 (adult vial test). After application of different concentrations of the insecticides were statistically evaluated their percent-age efficacy, degree of resistance of the population and lethal dose by Polo Plus 2.0 software. Based on the results, it was found that Brassicogethes aeneus populations showed resistance to lambda-cyhalothrin and tau-fluvalinate. Individuals were highly sensitive to indoxacarb. Brassicogethes aeneus showed considerable variability in the degrees of resistance against Biscaya 240 OD. Indoxacarb appears momentally to be the best way of plant protection, but further monitoring is important.
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Rezistence brouků škodících na obilninách k vybraným účinným látkám insekticidů
Benda, Ivo
The thesis titled “The Resistance of Beetles Harmful to Cereals Towards Selected Active Substances of Insecticides” deals with the issue of sensitivity of the oulema cereal leaf beetles towards active substances from the pyrethroid category (lambda-cyhalothrin) and organophosphates (chlorpyrifos-ethyl). Research was performed at a number of locations in the region of South Moravia. The beetles were collected on fields with common wheat (Triticum aestivum) and barley (Hordeum vulgare). The testing of the beetles was performed in a lab using the adult-vial test. Imagos were exposed to four concentrations of each active substance in three repetitions. The results were processed using the Polo Plus program. The activity of the beetles differed depending on the concentration of the active substance.
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Rezistence blýskáčka řepkového (Meligethes aeneus, Fabr. 1775) k insekticidům
Jetmarová, Zuzana
The aim of this thesis was to find the current occurrence of resistant sub-population of pollen beetle (Meligethes aeneus Fabricius, 1775) to the selected species active ingredients and the preparation. Adults of pollen beetles were collected from commonly used fields of oilseed rape plants at the time of the first process of flowering until the end of the entire flowering process. The sample collection was carried out during years 2012 and 2013 in twenty locations in the Czech Republic. The Levels of resistance were assessed in a laboratory test called Adult-vial- Test according to official methods used by the International Resistance Action Committee (IRAC). The first part of this thesis describes the biology of pollen beetles. It also describes other pests found on the oilseed rape plant. It further explains the basic concepts and summarizes the occurrence of resistant sub-populations of Pollen beetle in the Czech Republic and other European countries. The second part of this thesis constitutes the practical aspect of the research where there is testing of the resistance of sub-populations of pollen beetles on used active ingredients and the preparation of Biscaya 240 OD.
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Occurrence od vancomycin - resistant enterococci (VRE) in Tábor hospital, a.s.
NEKOLOVÁ, Tereza
This bachelor thesis deals with the occurrence of vancomycin-resistant enterococci in Tábor Hospital, a.s. The theoretical part of this bachelor thesis is focused on the basic information about the genus Enterococcus, the development of antibiotic resistence, the possibilities of diagnosing vancomycin-resistant enterococci and monitoring the development of this resistence. It also contains information on healthcare associated infections and prevention against them. The practical part describes the laboratory procedure of the examination of enterococci, their identification, determination of susceptibility to antimicrobial substances and subsequent determination of vancomycin resistance. Moreover, the practical part evaluates the obtained data on the occurrence of vancomycin-resistant enterococci in the Hospital Tábor, a.s. in the period from 2018 to 2021. The presence of vancomycin-resistant enterococci was evaluated within individual departments at Tábor Hospital, a.s. The percentage was compared with other enterococci, in the incidence and treatment of Clostridial colitis with vancomycin. Data from the whole nation were also used and evaluated. This thesis is focused on answering questions regarding the increased growth of vancomycin-resistant enterococci in previous years in the Tábor Hospital, a.s. as well as widely within the Czech Republic. Other question includes the increased incidence of vancomycin-resistant enterococci with the incidence and further treatment of Clostridial infection, if the same increasing trend occurs. The conclusion of the thesis is a summary of the importance of vancomycin-resistant enterococci, their frequency of occurrence according to the set goals and research questions, which have been answered.
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Výskyt vnitřních parazitů koní ve vybraných chovech
HOŘEJŠOVÁ, Barbora
The aim of the bachelor thesis was to process literature data on the occurrence, deve-lopmental stages and the possibilities of breeding measures aimed at reducing the occurrence of gastrointestinal parasites in horses, supplemented by self-monitoring in the specified breeding. The EPG values of the box housing were compared with a new only beginning type of active housing. Age, pedigree and climatic conditions were also taken into ac-count.
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Genotypization of selected candidate markers for resistance against lentiviral infections of small ruminants
FARKOVÁ, Barbora
Small Ruminant Lentiviruses (SRLV) known as maedi-visna in sheep, and viral arthritis and encephalitis in goats are widespread worldwide and cause significant economic losses. Lentiviral diseases have been included in the current Animal Health Control and Vaccination Guidelines for Breeding Controls. Control programs are generally based on serological screening and subsequently serologically positive animals are removed from breeding. In the literature review I write about important lentiviruses, including those that have not been analyzed. The aim of this work was to screen for the occurrence of lentiviral infections in small ruminants in the Czech Republic and the genotyping of selected candidate markers and finding the genotype related to the possibility of gene resistance. In total, over 3200 blood samples were collected from sheep and goats from breeders all over the Czech Republic. Genotyping was carried out in herds where serologically positive animals were detected. Amplification of the DNA sequence of interest was performed by PCR (Polymerase Chain Reaction) analysis, followed by genotyping by sequence analysis, and the results were statistically evaluated. In my work, I came to the conclusion that in the Czech Republic the occurrence of 2 out of 5 possible SRLV genotypes (genotype A and B) and distribution of different subtypes in sheep and goats has been detected so far. The greatest differences were in the sequence of genotype A, where one group of isolates showed the possibility of classification into a completely new subtype. In sheep, all three genotypes of the TMEM154 gene were identified. The incidence of serologically positive was statistically significantly higher in animals with a risk genotype. Because of the possible mutation of the virus and the emergence of a new subtype of genotype A, the virus is more likely to adapt to attack and infect individuals with a more resistant genotype. Only one genotype has been demonstrated in goats, so most likely there is no marker associated with resistance to SRLV infections.
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Interspecific varieties of grapevine and their utilization in the breeding
VOCÍLKA, Radek
In the thesis new interspecific varieties, which are in acknowledge-proceeding, have been observed. These varieties are Marlen, Cerason, Kofranka and Kaberon. These were cultivated from commonly used European varieties (Frankovka, Svatovavřinecké, Merlot) registrated in the State Book of Varieties of the Czech Republic. Moreover, during the cultivation blue variety called Seibel was used. This served as a donor of fungicidal desease resistance, or rather tollerance. The varieties were observed for three years and their morphological and degree qualities were investigated. The goal of the thesis was acknowledgement of these interspecific vine varieties, their registration in the State Book of Varieties and later legal protection. The task was to pass diversity, uniformity and stability tests in the proceeding and compare the new varieties with the acknowledged ones, which were used for cultivation.
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Impact of new FLT3 inhibitors on daunorubicin accumulation in ABCB1-expressing leukemic cells
Králová, Adéla ; Čečková, Martina (advisor) ; Jirkovský, Eduard (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Adéla Králová Supervisor: doc. PharmDr. Martina Čečková, Ph.D. Consultant: Mgr. Simona Suchá Title of Diploma thesis: Impact of new FLT3 inhibitors on daunorubicin accumulation in ABCB1-expressing leukemic cells Fms-like tyrosine kinase 3 (FLT3) inhibitors represent a new generation of drugs in the treatment of acute myeloid leukemia (AML). Standard therapeutic regimen of AML is initiated with induction therapy consisting of cytarabine and anthracyclines. The disadvantage of this combination is emerging resistance often caused by the ABCB1-mediated efflux. Therefore, simultaneous inhibition of FLT3 and ABCB1, which is inhibited by FLT3 inhibitors used in clinical practice, appears to be a beneficial approach to therapy. However, their effectiveness is declining hence the effort to develop new FLT3-inhibiting molecules. The aim of our work was to evaluate whether our two promising new FLT3-inhibiting compounds would inhibit ABCB1 as well. Promyelocyte cells overexpressing ABCB1 (HL60-ABCB1) and parent HL60-par were used in this study alongside AML-derived cell lines (MOLM-13, THP-1, Kasumi-1). Employing accumulation studies on HL60-ABCB1, strong inhibitory effect towards ABCB1 was demonstrated...
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