Despite numerous studies on Escherichia coli (E. coli) from sheep, there have been few reports on the characterization of E. coli isolates from various organs of individual sheep until now. The present study conducted molecular typing, antibiotics resistance, biofilm formation, and virulence genes on E. coli isolated from 57 freshly slaughtered apparently healthy sheep carcasses, gallbladders, fecal samples, and mesenteric lymph nodes (MLNs). The results demonstrated that the detection rate of R1 LPS core type in E. coli isolated from fecal samples (70.83%) was higher than that from other organs, but the detection rate of antibiotic resistance genes was lower (P < 0.05). The predominant phylogenetic group of E. coli isolated from the carcasses was group B1 (93.33%), and the detection rate of multidrug-resistance phenotype (80%) and the resistance rate of E. coli was higher than that from other organs (P < 0.05). Interestingly, the intensity of biofilm formation of E. coli isolated from MLNs was higher than that from other organs (P < 0.05). However, except for ibeB, the detection rates of virulence genes did not differ in E.coli isolated from different organs. In conclusion, differences were noted in these parameters of E. coli isolated from different organs of individual sheep. Therefore, the data may contain considerable mistakes concerning the actual situation in the host if we only analyze the data of E. coli isolated from feces or carcasses.

• MeSH
• antibakteriální látky * farmakologie   MeSH
• bakteriální léková rezistence genetika   MeSH
• biofilmy * růst a vývoj   účinky léků   MeSH
• Escherichia coli * genetika   účinky léků   izolace a purifikace   klasifikace   fyziologie   MeSH
• faktory virulence * genetika   MeSH
• feces mikrobiologie   MeSH
• fylogeneze MeSH
• infekce vyvolané Escherichia coli mikrobiologie   veterinární   MeSH
• molekulární typizace MeSH
• nemoci ovcí mikrobiologie   MeSH
• ovce MeSH
• virulence genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

INTRODUCTION: Hospitals and wastewater are recognized hot spots for the selection and dissemination of antibiotic-resistant bacteria to the environment, but the total participation of hospitals in the spread of nosocomial pathogens to municipal wastewater treatment plants (WWTPs) and adjacent rivers had not previously been revealed. METHODS: We used a combination of culturing and whole-genome sequencing to explore the transmission routes of Escherichia coli from hospitalized patients suffering from urinary tract infections (UTI) via wastewater to the environment. Samples were collected in two periods in three locations (A, B, and C) and cultured on selective antibiotic-enhanced plates. RESULTS: In total, 408 E. coli isolates were obtained from patients with UTI (n=81), raw hospital sewage (n=73), WWTPs inflow (n=96)/outflow (n=106), and river upstream (n=21)/downstream (n=31) of WWTPs. The majority of the isolates produced extended-spectrum beta-lactamase (ESBL), mainly CTX-M-15, and showed multidrug resistance (MDR) profiles. Seven carbapenemase-producing isolates with GES-5 or OXA-244 were obtained in two locations from wastewater and river samples. Isolates were assigned to 74 different sequence types (ST), with the predominance of ST131 (n=80) found in all sources including rivers. Extraintestinal pathogenic lineages frequently found in hospital sewage (ST10, ST38, and ST69) were also found in river water. Despite generally high genetic diversity, phylogenetic analysis of ST10, ST295, and ST744 showed highly related isolates (SNP 0-18) from different sources, providing the evidence for the transmission of resistant strains through WWTPs to surface waters. DISCUSSION: Results of this study suggest that 1) UTI share a minor participation in hospitals wastewaters; 2) a high diversity of STs and phylogenetic groups in municipal wastewaters derive from the urban influence rather than hospitals; and 3) pathogenic lineages and bacteria with emerging resistance genotypes associated with hospitals spread into surface waters. Our study highlights the contribution of hospital and municipal wastewater to the transmission of ESBL- and carbapenemase-producing E. coli with MDR profiles to the environment.

• MeSH
• antibakteriální látky farmakologie   MeSH
• beta-laktamasy genetika   MeSH
• Escherichia coli genetika   MeSH
• fylogeneze MeSH
• infekce močového ústrojí * mikrobiologie   MeSH
• infekce vyvolané Escherichia coli * mikrobiologie   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• multilokusová sekvenční typizace MeSH
• nemocnice MeSH
• odpadní voda MeSH
• odpadní vody mikrobiologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

OBJECTIVES: The beta-lactamases with extended spectrum of activity (ESBL) are medically one of the most important group of enzymes. Another group of beta-lactamases representing of Enterobacteriaceae is group of the AmpC-type cephalosporinases. The presented study provides identification and determination of the spectrum of resistance against different and clinically used antimicrobial drugs in the clinical isolates of Escherichia coli. METHODS: These isolates had origin in different departments of the L. Pasteur University Hospital in Košice. The goal was the detection of beta-lactamase production with extended-spectrum effect and testing of AmpC-type cephalosporinases by several phenotypic tests in clinical isolates. MALDI-TOF MS analysis was performed on a Microflex MALDI Biotyper. Samples were positively tested for ESBL with the use of the disc diffusion method. PCR were performed with a series of primers designed for the detection of Ambler class A, B and C beta-lactamase genes. RESULTS: For all 485 isolates, we determined the production of ESBL, which we detected in 166 E. coli isolates, which represents a 34.2% prevalence of ESBL production. It is clear from the results that the prevalence of ESBL-producing E. coli out of the total number of E. coli investigated reached 34.2%. In the monitored period, we confirmed at least one resistance gene from 485 E. coli in 188 positive isolates. CONCLUSIONS: We describe a complex ESBL epidemiology. The study revealed a high rate of ESBL-producing E. coli isolates; blaTEM and blaSHV enzymes dominated in ESBL-positive E. coli isolates in the L. Pasteur University Hospital in Košice.

• MeSH
• antibakteriální látky farmakologie   MeSH
• antibiotická rezistence MeSH
• Bacteria MeSH
• beta-laktamasy genetika   farmakologie   MeSH
• Escherichia coli * genetika   MeSH
• infekce vyvolané Escherichia coli * epidemiologie   mikrobiologie   MeSH
• lidé MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Routinely used typing methods including MLST, rep-PCR and whole genome sequencing (WGS) are time-consuming, costly, and often low throughput. Here, we describe a novel mini-MLST scheme for Eschericha coli as an alternative method for rapid genotyping. Using the proposed mini-MLST scheme, 10,946 existing STs were converted into 1,038 Melting Types (MelTs). To validate the new mini-MLST scheme, in silico analysis was performed on 73,704 strains retrieved from EnteroBase resulting in discriminatory power D = 0.9465 (CI 95% 0.9726-0.9736) for mini-MLST and D = 0.9731 (CI 95% 0.9726-0.9736) for MLST. Moreover, validation on clinical isolates was conducted with a significant concordance between MLST, rep-PCR and WGS. To conclude, the great portability, efficient processing, cost-effectiveness, and high throughput of mini-MLST represents immense benefits, even when accompanied with a slightly lower discriminatory power than other typing methods. This study proved mini-MLST is an ideal method to screen and subgroup large sets of isolates and/or quick strain typing during outbreaks. In addition, our results clearly showed its suitability for prospective surveillance monitoring of emergent and high-risk E. coli clones'.

• MeSH
• bakteriální geny * MeSH
• denaturace nukleových kyselin MeSH
• DNA bakterií chemie   genetika   MeSH
• DNA primery MeSH
• epidemický výskyt choroby MeSH
• Escherichia coli klasifikace   genetika   izolace a purifikace   MeSH
• genom bakteriální MeSH
• genotypizační techniky * MeSH
• infekce vyvolané Escherichia coli mikrobiologie   MeSH
• jednonukleotidový polymorfismus * MeSH
• multilokusová sekvenční typizace metody   MeSH
• počítačová simulace MeSH
• polymerázová řetězová reakce metody   MeSH
• repetitivní sekvence nukleových kyselin MeSH
• sekvenování celého genomu MeSH
• surveillance populace MeSH
• techniky typizace bakterií * MeSH
• zastoupení bazí MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

Intra-amniotic infections (IAI) are one of the reasons for preterm birth. High mobility group box 1 (HMGB1) is a nuclear protein with various physiological functions, including tissue healing. Its excessive extracellular release potentiates inflammatory reaction and can revert its action from beneficial to detrimental. We infected the amniotic fluid of a pig on the 80th day of gestation with 1 × 104 colony forming units (CFUs) of E. coli O55 for 10 h, and evaluated the appearance of HMGB1, receptor for glycation endproducts (RAGE), and Toll-like receptor (TLR) 4 in the amniotic membrane and fluid. Sham-infected amniotic fluid served as a control. The expression and release of HMGB1 were evaluated by Real-Time PCR, immunofluorescence, immunohistochemistry, and ELISA. The infection downregulated HMGB1 mRNA expression in the amniotic membrane, changed the distribution of HMGB1 protein in the amniotic membrane, and increased its level in amniotic fluid. All RAGE mRNA, protein expression in the amniotic membrane, and soluble RAGE level in the amniotic fluid were downregulated. TLR4 mRNA and protein expression and soluble TLR4 were all upregulated. HMGB1 is a potential target for therapy to suppress the exaggerated inflammatory response. This controlled expression and release can, in some cases, prevent the preterm birth of vulnerable infants. Studies on suitable animal models can contribute to the development of appropriate therapy.

• MeSH
• amnion imunologie   mikrobiologie   patologie   MeSH
• Escherichia coli růst a vývoj   patogenita   MeSH
• infekce vyvolané Escherichia coli genetika   imunologie   mikrobiologie   veterinární   MeSH
• infekční komplikace v těhotenství genetika   imunologie   mikrobiologie   veterinární   MeSH
• interakce hostitele a patogenu genetika   imunologie   MeSH
• lidé MeSH
• messenger RNA genetika   imunologie   MeSH
• modely nemocí na zvířatech MeSH
• plodová voda imunologie   mikrobiologie   MeSH
• prasata MeSH
• předčasný porod prevence a kontrola   MeSH
• protein HMGB1 genetika   imunologie   MeSH
• receptor pro konečné produkty pokročilé glykace genetika   imunologie   MeSH
• regulace genové exprese MeSH
• signální transdukce MeSH
• těhotenství MeSH
• toll-like receptor 4 genetika   imunologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Enterotoxigenic Escherichia coli (ETEC) and Shiga toxin-producing E. coli (STEC) strains are the causative agents of severe foodborne diseases in both humans and animals. In this study, porcine pathogenic E. coli strains (n = 277) as well as porcine commensal strains (n = 188) were tested for their susceptibilities to 34 bacteriocin monoproducers to identify the most suitable bacteriocin types inhibiting porcine pathogens. Under in vitro conditions, the set of pathogenic E. coli strains was found to be significantly more susceptible to the majority of tested bacteriocins than commensal E. coli. Based on the production of bacteriocins with specific activity against pathogens, three potentially probiotic commensal E. coli strains of human origin were selected. These strains were found to be able to outcompete ETEC strains expressing F4 or F18 fimbriae in liquid culture and also decreased the severity and duration of diarrhea in piglets during experimental ETEC infection as well as pathogen numbers on the last day of in vivo experimentation. While the extents of the probiotic effect were different for each strain, the cocktail of all three strains showed the most pronounced beneficial effects, suggesting synergy between the tested E. coli strains. IMPORTANCE Increasing levels of antibiotic resistance among bacteria also increase the need for alternatives to conventional antibiotic treatment. Pathogenic Escherichia coli represents a major diarrheic infectious agent of piglets in their postweaning period; however, available measures to control these infections are limited. This study describes three novel E. coli strains producing antimicrobial compounds (bacteriocins) that actively inhibit a majority of toxigenic E. coli strains. The beneficial effect of three potentially probiotic E. coli strains was demonstrated under both in vitro and in vivo conditions. The novel probiotic candidates may be used as prophylaxis during piglets' postweaning period to overcome common infections caused by E. coli.

• MeSH
• bakteriální toxiny * metabolismus   MeSH
• bakteriociny metabolismus   terapeutické užití   MeSH
• Escherichia coli * účinky léků   genetika   metabolismus   MeSH
• faktory virulence genetika   MeSH
• feces mikrobiologie   MeSH
• infekce vyvolané Escherichia coli mikrobiologie   prevence a kontrola   veterinární   MeSH
• nemoci prasat mikrobiologie   prevence a kontrola   MeSH
• prasata MeSH
• probiotika terapeutické užití   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• randomizované kontrolované studie veterinární MeSH

Shiga toxin-producing Escherichia coli (STEC) is an important foodborne pathogens, known to cause enteric infections especially diarrhea, mainly attributed to Shiga toxins (Stxs). The use of certain antibiotics for treating this infection is controversial, owing to an increased risk for producing Stxs (Stx 1 and Stx 2). Increased antibiotic resistance is also thought to be involved in the pathogenesis of STEC diseases. The purpose of this study was to analyze the effects of antibiotics on induction of Stx 1 and Stx 2 in clinical STEC isolates and to investigate the relationships between increased resistance and Stx production. Fifteen clinical isolates were treated with sub minimum inhibitory concentrations (Sub MIC) of clinically used antibiotics (ciprofloxacin, fosfomycin, tigecycline, and meropenem), and the changes in expression levels of stx1 and stx2 genes were estimated using qRT-PCR. The expressions of Shiga toxins were found to be increased up to 6.5- and eightfold under ciprofloxacin and tigecycline Sub MIC, respectively. Fosfomycin had weak induction effect of up to twofold, whereas meropenem had the weakest influence on such expression. Resistant isolates were found to be more prone to increased expression of toxins.

• MeSH
• antibakteriální látky farmakologie   MeSH
• infekce vyvolané Escherichia coli mikrobiologie   MeSH
• lidé MeSH
• regulace genové exprese u bakterií * účinky léků   MeSH
• shiga toxin 1 * genetika   MeSH
• shiga toxin 2 * genetika   MeSH
• shiga-toxigenní Escherichia coli * účinky léků   genetika   MeSH
• stanovení celkové genové exprese MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Upsurge in the instances of antibiotic-resistant uropathogenic Escherichia .coli (UPECs) strains has repositioned the attention of researchers towards a century old antimicrobial approach popularly known as phage therapy. Rise of extended spectrum beta lactamase (ESBL) and biofilm producing strains has added another step of hurdle in treatment of uropathogens with conventional antibiotics, thus providing a further impetus for search for exploring new therapeutic measures. In this direction, bacteriophages, commonly called phages, are recently being considered as potential alternatives for treatment of UPECs. Phages are the tiniest form of viruses which are ubiquitous in nature and highly specific for their host. This review discusses the possible ways of using natural phages, genetically engineered phages, and phage lytic enzymes (PLEs) as an alternative antimicrobial treatment for urinary tract infections. The review also sheds light on the synergistic use of conventional antibiotics with phages or PLEs for treatment of uropathogens. These methods of using phages and their derivatives, alone or in combination with antibiotics, have proved fruitful so far in in vitro studies. However, in vivo studies are required to make them accessible for human use. The present review is a concerted effort towards putting together all the information available on the subject.

• MeSH
• bakteriofágy genetika   fyziologie   MeSH
• fágová terapie * MeSH
• infekce močového ústrojí mikrobiologie   terapie   MeSH
• infekce vyvolané Escherichia coli mikrobiologie   terapie   MeSH
• lidé MeSH
• uropatogenní Escherichia coli genetika   fyziologie   virologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

We evaluated the prevalence and epidemiology of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli isolates in pigs during production cycle on a Czech farm with the history of previous use of ceftiofur. ESBL-producing E. coli isolates were obtained from rectal swabs from pigs of different age groups (suckling piglets, weaned piglets, growers and sows). Collected samples were directly cultivated on MacConkey agar with cefotaxime (2 mg l-1 ), whereas intestinal swabs of slaughtered pigs and surface swabs from pig carcasses were also pre-enriched in buffered peptone water without antimicrobials before the cultivation. Clonal relationship of selected isolates was determined by XbaI pulse-field gel electrophoresis and multi-locus sequence typing. The transferability of plasmids carrying blaCTX-M genes was tested by conjugation experiments. From all examined samples, 141 (43·7%, n = 323) were positive for ESBL-producing E. coli. All ESBL-producing isolates showed resistance to multiple antimicrobials and were positive for blaCTX-M genes. The blaCTX-M-1 was carried by conjugative IncN/ST1 plasmids (c. 40-45 kb) while the blaCTX-M-15 was located on conjugative F plasmids with F:18:A5:B1 formula (c. 165 kb). This study demonstrated the persistence of CTX-M-positive E. coli isolates 2 months after banner of ceftiofur usage and indicated possible risk of transmission of these isolates to humans via the food chain.

• MeSH
• beta-laktamasy genetika   metabolismus   MeSH
• Escherichia coli enzymologie   genetika   izolace a purifikace   MeSH
• farmy MeSH
• infekce vyvolané Escherichia coli mikrobiologie   patofyziologie   veterinární   MeSH
• lidé MeSH
• multilokusová sekvenční typizace MeSH
• nemoci prasat mikrobiologie   patofyziologie   MeSH
• plazmidy genetika   metabolismus   MeSH
• prasata růst a vývoj   mikrobiologie   MeSH
• proteiny z Escherichia coli genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

BACKGROUND: Avian pathogenic Escherichia coli (APEC) can cause various extraintestinal infections in poultry, resulting in massive economic losses in poultry industry. In addition, some avian E. coli strains may have zoonotic potential, making poultry a possible source of infection for humans. Due to its extreme genetic diversity, this pathotype remains poorly defined. This study aimed to investigate the diversity of colibacillosis-associated E. coli isolates from Central European countries with a focus on the Czech Republic. RESULTS: Of 95 clinical isolates subjected to preliminary characterization, 32 were selected for whole-genome sequencing. A multi resistant phenotype was detected in a majority of the sequenced strains with the predominant resistance to β-lactams and quinolones being associated with TEM-type beta-lactamase genes and chromosomal gyrA mutations respectively. The phylogenetic analysis confirmed a great diversity of isolates, that were derived from nearly all phylogenetic groups, with predominace of B2, B1 and C phylogroups. Clusters of closely related isolates within ST23 (phylogroup C) and ST429 (phylogroup B2) indicated a possible local spread of these clones. Besides, the ST429 cluster carried blaCMY-2, - 59 genes for AmpC beta-lactamase and isolates of both clusters were generally well-equipped with virulence-associated genes, with considerable differences in distribution of certain virulence-associated genes between phylogenetically distant lineages. Other important and potentially zoonotic APEC STs were detected, incl. ST117, ST354 and ST95, showing several molecular features typical for human ExPEC. CONCLUSIONS: The results support the concept of local spread of virulent APEC clones, as well as of zoonotic potential of specific poultry-associated lineages, and highlight the need to investigate the possible source of these pathogenic strains.

• MeSH
• Escherichia coli klasifikace   genetika   izolace a purifikace   patogenita   MeSH
• fylogeneze MeSH
• genetická variace MeSH
• infekce vyvolané Escherichia coli mikrobiologie   veterinární   MeSH
• kur domácí MeSH
• nemoci drůbeže mikrobiologie   MeSH
• sekvenování celého genomu MeSH
• virulence genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH