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Antimicrobial resistance in clinical isolates of Clostridioides difficile derived from hospitalised patients in the Czech Republic and Slovakia.
Zíková, Jaroslava ; Krůtová, Marcela (advisor) ; Najmanová, Lucie (referee)
Clostridioides difficile is one of the most common pathogens of healthcare-associated colitis (CDI). However, in recent years there has been an increasing number of reported cases in the community. Causative strain characterization allows us to track the occurrence and spread of certain C. difficile types. To update the current CDI epidemiological situation, between October and November 2021, 342 C. difficile isolates or stool samples from CDI patients for culture were sent from 23 Czech (n=200) and 17 Slovak (n=142) hospitals. All C. difficile isolates were characterized by ribotyping and toxin genes detection. From 342 isolates, 52 different ribotyping profiles were identified. The frequent ribotypes detected were 001, 176, 014, 018 and 020. Genes for toxins A and B were found in all isolates. Genes for binary toxins were detected in 43 (21,5 %) Czech isolates and in 117 (82,4 %) Slovak isolates. Further, selected isolates (n=140, 40,9 %) were subjected to whole-genome sequencing (WGS) and antimicrobial susceptibility testing to 17 antibiotics (E-test). Multilocus sequencing typing determined 30 different sequence types from the 39 included ribotypes. All C. difficile isolates were susceptible to vancomycin, tetracycline, amoxicillin, and meropenem. The most frequently detected antimicrobial...
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A comparative analysis of the isolates of Clostridioides difficile derived from different sources.
Eretová, Veronika ; Krůtová, Marcela (advisor) ; Ježek, Petr (referee)
Clostridioides difficile infections (CDI) are generally perceived as healtcare associated infections. However, there has been recently reported an increase of CDI incidence in the community. The occurrence of C. difficile has been described also in animals, food and the environment water and soil. The aim of this thesis was to characterize C. difficile isolates derived from different sources using molecular methods. The results were discussed with available data from Czech human C. difficile isolates. A total of 135 C. difficile isolates from the following sources were analyzed: pigs n = 57, calves n = 44, horses n = 18, water n = 15 and hedgehog. Using PCR ribotyping, 22 distinct ribotyping profiles were identified, the most frequently detected ribotypes were: 033, 011, 126, 078. Both toxigenic and non-toxigenic ribotypes were detected, including binary toxin-producing strains. The most frequently detected antimicrobial resistances were to ciprofloxacin, clindamycin, and erythromycin. All ribotypes and sequence types identified in the C. difficile isolates from animals and the environment, has been found also in C. difficile isolates from humans which confirms the role of animals and the environment as a source for C. difficile. However, no epidemic ribotype 001 and 176 that dominate the current...
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Clostridium difficile: Molecular typing of clinically significant isolates
Krůtová, Marcela ; Nyč, Otakar (advisor) ; Čermák, Pavel (referee) ; Krásný, Libor (referee)
Currently, Clostridium difficile is a leading nosocomial pathogen due to the spread of epidemic strains. Molecular typing of clinical isolates is an important part of C. difficile occurrence and spread control in hospitals as well as in the community. A total of 2201 clinical C. difficile isolates from 32 hospitals cultured between 2013-2015 were characterized by PCR ribotyping and toxin gene multiplex PCR. A total of 166 different ribotyping profiles were identified, of which 53 ribotyping profiles were represented by at least two isolates for each profile. The most frequently found ribotypes were 176 (n=588, 26.7%) and 001 (n=456, 20.7%) followed by 014 (n=176, 8%), 012 (n=127, 5.8%), 017 (n=85, 3.9%) and 020 (n=68, 3.1%). Out of 2201 isolates, 2024 (92%) isolates were toxigenic and carried genes for toxin A and B, and of these, 677 (33.5%) also carried genes for binary toxin. The remaining 177 (8%) isolates were non-toxigenic. Subtyping of C. difficile isolates using a multilocus variable-number tandem repeats analysis (MLVA), that compared the sum of tandem repeats differences, was performed in C. difficile isolates of ribotype 176 (n=225, 17 hospitals) and in C. difficile isolates of ribotype 001 (n=184, 14 hospitals) cultured in 2014. The clonal relatedness in C. difficile isolates belonging...
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