Although Campylobacter jejuni is the pathogen responsible for the most common foodborne illness, tracing of the infection source remains challenging due to its highly variable genome. Therefore, one of the aim of the study was to compare three genotyping methods (MLST, PFGE, and mP-BIT) to determine the most effective genotyping tool. C. jejuni strains were divided into 4 clusters based on strain similarity in the cgMLST dendrogram. Subsequently, the dendrograms of the 3 tested methods were compared to determine the accuracy of each method compared to the reference cgMLST method. Moreover, a cost-benefit analysis has showed that MLST had the highest inverse discrimination index (97%) and required less workflow, time, fewer consumables, and low bacterial sample quantity. PFGE was shown to be obsolete both because of its low discriminatory power and the complexity of the procedure. Similarly, mP‐BIT showed low separation results, which was compensated by its high availability. Therefore, our data showed that MLST is the optimal tool for genotyping C. jejuni. Another aim was to compare the antimicrobial resistance to ciprofloxacin, erythromycin, and tetracycline in C. jejuni strains isolated from human, water, air, food, and animal samples by two gene sequence-based prediction methods and to compare them with the actual susceptibility of C. jejuni strains using the disc diffusion method. Both tools, ResFinder and RGI, synchronously predict the antimicrobial susceptibility of C. jejuni and either can be used.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence genetika MeSH
- Campylobacter jejuni * genetika MeSH
- genotyp MeSH
- kampylobakterové infekce * mikrobiologie MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- multilokusová sekvenční typizace MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Publikační typ
- abstrakt z konference MeSH
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x
- MeSH
- kontrola potravin MeSH
- lidé MeSH
- potravní doplňky MeSH
- probiotika * MeSH
- Check Tag
- lidé MeSH
Antimicrobial resistance (AMR) is one of the top public health threats nowadays. Among the most important AMR pathogens, Escherichia coli resistant to extended spectrum cephalosporins (ESC-EC) is a perfect example of the One Health problem due to its global distribution in animal, human, and environmental sources and its resistant phenotype, derived from the carriage of plasmid-borne extended-spectrum and AmpC β-lactamases, which limits the choice of effective antimicrobial therapies. The epidemiology of ESC-EC infection is complex as a result of the multiple possible sources involved in its transmission, and its study would require databases ideally comprising information from animal (livestock, companion, wildlife), human, and environmental sources. Here, we present the steps taken to assemble a database with phenotypic and genetic information on 10,763 ESC-EC isolates retrieved from multiple sources provided by 13 partners located in eight European countries, in the frame of the DiSCoVeR Joint Research project funded by the One Health European Joint Programme (OH-EJP), along with its strengths and limitations. This database represents a first step to help in the assessment of different geographical and temporal trends and transmission dynamics in animals and humans. The work performed highlights aspects that should be considered in future international efforts, such as the one presented here.
- Publikační typ
- časopisecké články MeSH
x
x
- MeSH
- analýza potravin MeSH
- epidemiologické monitorování MeSH
- lidé MeSH
- methicilin rezistentní Staphylococcus aureus * genetika klasifikace patogenita MeSH
- přenos infekční nemoci prevence a kontrola MeSH
- prevalence MeSH
- techniky typizace bakterií MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
- Geografické názvy
- Česká republika MeSH
Cíl práce: Cílem studie bylo určit genetickou rozmanitost humánních izolátů získaných v letech 2016–2020 z klinických laboratoří z různých lokalit České republiky (ČR) s ohledem na možné epidemické souvislosti a virulenci Listeria monocytogenes za využití dat celogenomového sekvenování. Metody: Ve studii byl použit soubor 102 humánních izolátů L. monocytogenes, u kterých byl určen sérotyp sklíčkovou aglutinací v kombinaci s multiplex PCR sérotypizací. Retrospektivně bylo provedeno celogenomové sekvenování a na základě získaných dat byla posouzena klonální příbuznost testovaných kmenů a přítomnost genů virulence pomocí softwaru Ridom SeqSphere+. Výsledky: V období let 2016–2020 bylo celkem charakterizováno 102 humánních izolátů L. monocytogenes, což činí 65 % ze všech hlášených případů listerióz registrovaných ve sledovaném období v ČR v systémech ISIN/EPIDAT. Dominantní zastoupení měl sérotyp 1/2a (57 %), následoval sérotyp 4b (30 %) a jen ojediněle byly detekovány kmeny sérotypu 1/2b (12 %) a 1/2c (1 %). Na základě analýzy celogenomových dat byly kmeny zařazeny k 26 klonálním komplexům a 27 sekvenačním typům. Porovnáním cgMLST (core genome Multi-Locus Sequence Typing) byly detekovány čtyři klastry o více jak třech kmenech vykazující vysokou příbuznost (rozdíly do 10 alel) s možnou epidemickou souvislostí. U všech kmenů byla potvrzena přítomnost klíčových genů virulence. Pouze u tří kmenů (sérotypu 1/2a, 1/2b a 1/2c) byla detekována bodová mutace v genu inlA spojovaná s expresí zkráceného proteinu internalinu A, který je zapojený do mechanismu přestupu L. monocytogenes intestinální bariérou. Závěr: Molekulární epidemiologie založená na celogenomovém sekvenování představuje účinný nástroj ke studiu populační struktury kmenů L. monocytogenes. V této studii byla zjištěna vysoká heterogenita humánních kmenů L. monocytogenes, zejména u v České republice dominantního sérotypu 1/2a. Bylo identifikováno několik klastrů s možnou epidemickou souvislostí, jejichž výskyt bude dále sledován.
Study aim: To determine the genetic diversity of human isolates of Listeria monocytogenes obtained in 2016–2020 from clinical laboratories in various locations of the Czech Republic with a focus on their possible epidemic links and virulence using whole genome sequencing data. Methods: A total of 102 human L. monocytogenes isolates, serotyped by slide agglutination in combination with multiplex PCR serotyping, were used in this study. Whole genome sequencing was performed retrospectively, and based on the obtained data, the clonal relatedness of the tested strains and the presence of virulence genes were assessed using the Ridom SeqSphere+ software. Results: In 2016-2020, 102 human isolates of L. monocytogenes were characterized, which represented 65% of all cases of listeriosis reported to the ISIN/EPIDAT systems in the Czech Republic in the monitored period. Serotype 1/2a (57%) was dominant, followed by serotype 4b (30%). Strains of serotype 1/2b (12%) and 1/2c (1%) were rarely detected. Based on the analysis of whole genome sequencing data, the strains were assigned to 26 clonal complexes and 27 sequence types. The cgMLST (core genome Multi-Locus Sequence Typing) analysis revealed four clusters of more than three strains, showing high relatedness (differences up to 10 alleles) with a possible epidemic link. The presence of all key virulence genes was confirmed in all strains. Only three strains (of serotypes 1/2a, 1/2b, and 1/2c) carried a point mutation in the inlA gene responsible for the expression of truncated internalin A protein, which is involved in the mechanism of intestinal barrier crossing by L. monocytogenes. Conclusion: Molecular epidemiology based on whole genome sequencing is an effective tool to study the population structure of L. monocytogenes strains. This study found high heterogeneity of human L. monocytogenes strains, especially for serotype 1/2a, dominant in the Czech Republic. Several clusters with a possible epidemic link have been identified, and their occurrence will be further monitored.
This study was aimed on the detection of methicillin resistant Staphylococcus aureus (MRSA) in different categories of retailed ready-to-eat (RTE) meat products from the Czech producers and determination of their genetic properties, antimicrobial resistance and virulence. In RTE meat products, 2% (4/181) of examined samples were MRSA positive. MRSA strains were detected only in durable fermented meat products made exclusively from pork meat. Detection of livestock-associated MRSA (LA-MRSA) clonal lineages (ST398 and ST4999), SCCmec cassette type V and tetracycline resistance indicate a source of contamination from raw pork. The study confirms the ability of these strains to survive the technological process rather than contamination of meat products from the food processing environment. MRSA strains did not carry any of the tested genes encoding staphylococcal enterotoxins or virulence genes (for Panton-Valentine leukocidin, exfoliative toxins A, B and toxic shock syndrome). Our results point out the spread of LA-MRSA through the meat processing chain.
- MeSH
- antibakteriální látky farmakologie MeSH
- dobytek MeSH
- masné výrobky * MeSH
- maso MeSH
- methicilin rezistentní Staphylococcus aureus * genetika MeSH
- mikrobiální testy citlivosti MeSH
- stafylokokové infekce * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
Listeria monocytogenes (Lm) is a ubiquitous bacterium that causes listeriosis, a serious foodborne illness. In the nature-to-human transmission route, Lm can prosper in various ecological niches. Soil and decaying organic matter are its primary reservoirs. Certain clonal complexes (CCs) are over-represented in food production and represent a challenge to food safety. To gain new understanding of Lm adaptation mechanisms in food, the genetic background of strains found in animals and environment should be investigated in comparison to that of food strains. Twenty-one partners, including food, environment, veterinary and public health laboratories, constructed a dataset of 1484 genomes originating from Lm strains collected in 19 European countries. This dataset encompasses a large number of CCs occurring worldwide, covers many diverse habitats and is balanced between ecological compartments and geographic regions. The dataset presented here will contribute to improve our understanding of Lm ecology and should aid in the surveillance of Lm. This dataset provides a basis for the discovery of the genetic traits underlying Lm adaptation to different ecological niches.
BACKGROUND: Travellers were recognized as a risk cohort that can be colonized by mcr-1-mediated colistin-resistant Enterobacteriaceae. We aimed to investigate the carriage of mcr-mediated colistin resistance in Enterobacteriaceae in Czech travellers or expatriates residing temporarily in the Czech Republic. METHODS: Between August 2018 and September 2019, the stool samples were cultured in enrichment broth. The enriched cultures were tested for the presence of the mcr-1-8 genes and inoculated onto selective agar with colistin. Colistin-resistant Enterobacteriaceae were tested for the presence of the mcr-1-8 genes; the mcr-positive isolates were characterised by whole genome sequencing. RESULTS: From the 177 stool samples, 15 colistin-resistant Enterobacteriaceae isolates were cultured (7.9%); two of the E. coli isolates carried the mcr-1 gene (1.1%). In the E. coli multilocus sequence type (ST) 156, the mcr-1 gene was located in an ISApl1-mcr-1-orf-ISApl1 (Tn6330) and incorporated into the chromosome; in the E. coli ST23 isolate, the mcr-1 gene was harboured by the plasmid IncX4. Both of the mcr-1 positive E. coli isolates were multidrug-resistant and one isolate was an extended-spectrum β-lactamase producer (blaCTX-M-27). CONCLUSION: Patients with an international travel history should be monitored for the carriage of the mcr-1 gene in order to prevent its dissemination into healthcare settings.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence genetika MeSH
- chromozomy MeSH
- Enterobacteriaceae genetika MeSH
- Escherichia coli genetika MeSH
- kolistin * farmakologie MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- plazmidy genetika MeSH
- proteiny z Escherichia coli * genetika MeSH
- průřezové studie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
In August 2017, an increased incidence of Salmonella Bareilly was detected in the Czech Republic. An investigation was conducted with Slovakia to confirm the outbreak and identify the source. Probable outbreak cases were defined as cases with laboratory-confirmed S. Bareilly reported in either of the national surveillance systems, and/or the Czech and Slovak National Reference Laboratory databases from July 2017. Confirmed cases had the pulsed-field gel electrophoresis (PFGE) outbreak pulsotype or up to 5 alleles difference from outbreak cluster members by core genome multilocus sequence typing (cgMLST). PFGE and whole genome sequencing were used for isolate comparison. The same trawling questionnaire was used in both countries. By the end of October 2018, 325 cases were identified. Among 88 human S. Bareilly isolates analysed by PFGE, 82 (93%) shared an identical pulsotype; cgMLST of 17 S. Bareilly human isolates showed 1-2 allele difference. The trawling questionnaire excluded consumption of unusual or imported foods. In September 2018, an isolate closely related to the outbreak isolates was identified in a powdered egg product. A spray dryer was recognised as the contamination source and the production plant was closed. Using molecular typing methods, we detected a diffuse cross-border outbreak caused by S. Bareilly.
- MeSH
- epidemický výskyt choroby * MeSH
- genom bakteriální MeSH
- lidé MeSH
- multilokusová sekvenční typizace MeSH
- pulzní gelová elektroforéza MeSH
- Salmonella * genetika MeSH
- sekvenování celého genomu MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
- Slovenská republika MeSH
