Wild birds including raptors can act as vectors of clinically relevant bacteria with antibiotic resistance. The aim of this study was to investigate the occurrence of antibiotic-resistant Escherichia coli in black kites (Milvus migrans) inhabiting localities in proximity to human-influenced environments in southwestern Siberia and investigate their virulence and plasmid contents. A total of 51 E. coli isolates mostly with multidrug resistance (MDR) profiles were obtained from cloacal swabs of 35 (64%, n = 55) kites. Genomic analyses of 36 whole genome sequenced E. coli isolates showed: (i) high prevalence and diversity of their antibiotic resistance genes (ARGs) and common association with ESBL/AmpC production (27/36, 75%), (ii) carriage of mcr-1 for colistin resistance on IncI2 plasmids in kites residing in proximity of two large cities, (iii) frequent association with class one integrase (IntI1, 22/36, 61%), and (iv) presence of sequence types (STs) linked to avian-pathogenic (APEC) and extra-intestinal pathogenic E. coli (ExPEC). Notably, numerous isolates had significant virulence content. One E. coli with APEC-associated ST354 carried qnrE1 encoding fluoroquinolone resistance on IncHI2-ST3 plasmid, the first detection of such a gene in E. coli from wildlife. Our results implicate black kites in southwestern Siberia as reservoirs for antibiotic-resistant E. coli. It also highlights the existing link between proximity of wildlife to human activities and their carriage of MDR bacteria including pathogenic STs with significant and clinically relevant antibiotic resistance determinants. IMPORTANCE Migratory birds have the potential to acquire and disperse clinically relevant antibiotic-resistant bacteria (ARB) and their associated antibiotic resistance genes (ARGs) through vast geographical regions. The opportunistic feeding behavior associated with some raptors including black kites and the growing anthropogenic influence on their natural habitats increase the transmission risk of multidrug resistance (MDR) and pathogenic bacteria from human and agricultural sources into the environment and wildlife. Thus, monitoring studies investigating antibiotic resistance in raptors may provide essential data that facilitate understanding the fate and evolution of ARB and ARGs in the environment and possible health risks for humans and animals associated with the acquisition of these resistance determinants by wildlife.
- MeSH
- Angiotensin Receptor Antagonists MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Animals, Wild MeSH
- Escherichia coli * MeSH
- Angiotensin-Converting Enzyme Inhibitors MeSH
- Humans MeSH
- Drug Resistance, Multiple, Bacterial genetics MeSH
- Escherichia coli Proteins * genetics MeSH
- Birds microbiology MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Siberia MeSH
Drůbež a drůbeží produkty jsou považovány za potenciálně významný zdroj rezistentních kmenů E. coli a jejich determinant kódovaných mobilními elementy, jako jsou geny pro ESBL/AmpC betalaktamázy. Drůbež je také hostitelem kmenů s extraintestinální patogenitou (ExPEC). Podobnost drůbežích APEC s humánními kmeny vedla k zařazení ExPEC mezi bakterie se zoonotickým potenciálem. Populační diverzita různých patotypů E. coli, druhová specifita a cirkulace různých kmenů a genetických determinant mezi populacemi jsou však velmi složité a výše uvedená rizika nelze řešit odděleně jedno od druhého.
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- MeSH
- Bacterial Zoonoses MeSH
- Escherichia coli * metabolism pathogenicity MeSH
- Extraintestinal Pathogenic Escherichia coli MeSH
- Poultry Diseases nursing prevention & control MeSH
- Disease Transmission, Infectious MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
The great plasticity and diversity of the Escherichia coli genome, together with the ubiquitous occurrence, make E. coli a bacterium of world-wide concern. Of particular interest are pathogenic strains and strains harboring antimicrobial resistance genes. Overlapping virulence-associated traits between avian-source E. coli and human extraintestinal pathogenic E. coli (ExPEC) suggest zoonotic potential and safety threat of poultry food products. We analyzed whole-genome sequencing (WGS) data of 46 mcr-1-positive E. coli strains isolated from retail raw meat purchased in the Czech Republic. The investigated strains were characterized by their phylogroup-B1 (43%), A (30%), D (11%), E (7%), F (4%), B2 (2%), C (2%), MLST type, and serotype. A total of 30 multilocus sequence types (STs), of which ST744 was the most common (11%), were identified, with O8 and O89 as the most prevalent serogroups. Using the VirulenceFinder tool, 3 to 26 virulence genes were detected in the examined strains and a total of 7 (15%) strains met the pathogenic criteria for ExPEC. Four strains were defined as UPEC (9%) and 18 (39%) E. coli strains could be classified as APEC. The WGS methods and available on-line tools for their evaluation enable a comprehensive approach to the diagnosis of virulent properties of E. coli strains and represent a suitable and comfortable platform for their detection. Our results show that poultry meat may serve as an important reservoir of strains carrying both virulence and antibiotic resistance genes for animal and human populations.
- Publication type
- Journal Article MeSH
Silver gulls carry phylogenetically diverse Escherichia coli, including globally dominant extraintestinal pathogenic E. coli (ExPEC) sequence types and pandemic ExPEC-ST131 clades; however, our large-scale study (504 samples) on silver gulls nesting off the coast of New South Wales identified E. coli ST457 as the most prevalent. A phylogenetic analysis of whole-genome sequences (WGS) of 138 ST457 samples comprising 42 from gulls, 2 from humans (Australia), and 14 from poultry farmed in Paraguay were compared with 80 WGS deposited in public databases from diverse sources and countries. E. coli ST457 strains are phylogenetic group F, carry fimH145, and partition into five main clades in accordance to predominant flagella H-antigen carriage. Although we identified considerable phylogenetic diversity among the 138 ST457 strains, closely related subclades (<100 SNPs) suggested zoonotic or zooanthroponosis transmission between humans, wild birds, and food-producing animals. Australian human clinical and gull strains in two of the clades were closely related (≤80 SNPs). Regarding plasmid content, country, or country/source, specific connections were observed, including I1/ST23, I1/ST314, and I1/ST315 disseminating blaCMY-2 in Australia, I1/ST113 carrying blaCTX-M-8 and mcr-5 in Paraguayan poultry, and F2:A-:B1 plasmids of Dutch origin being detected across multiple ST457 clades. We identified a high prevalence of nearly identical I1/ST23 plasmids carrying blaCMY-2 among Australian gull and clinical human strains. In summary, ST457 is a broad host range, geographically diverse E. coli lineage that can cause human extraintestinal disease, including urinary tract infection, and displays a remarkable ability to capture mobile elements that carry and transmit genes encoding resistance to critically important antibiotics.
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- beta-Lactamases genetics MeSH
- beta-Lactams MeSH
- Animals, Wild MeSH
- Escherichia coli * genetics MeSH
- Phylogeny MeSH
- Escherichia coli Infections * veterinary MeSH
- Humans MeSH
- Plasmids genetics MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Australia 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 classification genetics isolation & purification pathogenicity MeSH
- Phylogeny MeSH
- Genetic Variation MeSH
- Escherichia coli Infections microbiology veterinary MeSH
- Chickens MeSH
- Poultry Diseases microbiology MeSH
- Whole Genome Sequencing MeSH
- Virulence genetics MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Czech Republic MeSH
A set of 178 Escherichia coli isolates taken from patients with inflammatory bowel disease (IBD) was analyzed for bacteriocin production and tested for the prevalence of 30 bacteriocin and 22 virulence factor determinants. Additionally, E. coli phylogenetic groups were also determined. Pulsed-field gel electrophoresis (PFGE) was used for exclusion of clonal character of isolates. Results were compared to data from a previously published analysis of 1283 fecal commensal E. coli isolates. The frequency of bacteriocinogenic isolates (66.9%) was significantly higher in IBD E. coli compared to fecal commensal E. coli isolates (54.2%, p < 0.01). In the group of IBD E. coli isolates, a higher prevalence of determinants for group B colicins (i.e., colicins B, D, Ia, Ib, M, and 5/10) (p < 0.01), including a higher prevalence of the colicin B determinant (p < 0.01) was found. Virulence factor determinants encoding fimbriae (fimA, 91.0%; pap, 27.5%), cytotoxic necrotizing factor (cnf1, 11.2%), aerobactin synthesis (aer, 43.3%), and the locus associated with invasivity (ial, 9.0%) were more prevalent in IBD E. coli (p < 0.05 for all five determinants). E. coli isolates from IBD mucosal biopsies were more frequently bacteriocinogenic (84.6%, p < 0.01) compared to fecal IBD isolates and fecal commensal E. coli. PFGE analysis revealed clusters specific for IBD E. coli isolates (n = 11), for fecal isolates (n = 13), and clusters containing both IBD and fecal isolates (n = 10). ExPEC (Extraintestinal Pathogenic E. coli) virulence and colicin determinants appear to be important characteristics of IBD E. coli isolates, especially the E. coli isolates obtained directly from biopsy samples.
- MeSH
- Bacterial Toxins genetics MeSH
- Bacteriocins metabolism MeSH
- Crohn Disease microbiology MeSH
- Escherichia coli genetics isolation & purification metabolism MeSH
- Extraintestinal Pathogenic Escherichia coli isolation & purification pathogenicity MeSH
- Humans MeSH
- Oxo-Acid-Lyases genetics MeSH
- Fimbriae Proteins genetics MeSH
- Escherichia coli Proteins genetics MeSH
- Electrophoresis, Gel, Pulsed-Field MeSH
- Gastrointestinal Microbiome physiology MeSH
- Colitis, Ulcerative microbiology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Antimikrobiální peptidy jsou významnou složkou nespecifické imunity téměř všech forem života a významně se podílí na obraně jedince proti patogenním mikroorganismům. Během jejich dlouholetého studia byly popsány fyzikálně‑chemické vlastnosti molekul peptidů, klíčové pro jejich antimikrobiální účinek (kationicita, hydrofobicita, amfipaticita), a pro mnohé byly navrženy různé mechanismy jejich působení. Počáteční fází působení kationických peptidů je elektrostatická interakce se záporně nabitým povrchem bakteriální buňky. V přítomnosti bakteriální membrány nabývají molekuly peptidů své amfipatické sekundární struktury umožňující postranním řetězcům jejich hydrofobních aminokyselin zanoření dovnitř fosfolipidové dvojvrstvy membrány. To má za následek narušení struktury membrány a/nebo vznik pórů. Dochází k rozvratu biologických funkcí membrány, což v samotném důsledku vede k zániku celé buňky. Účinek je rychlý a natolik odlišný od účinku antibiotik používaných v současné klinické praxi, že se obecně předpokládá, že by se antimikrobiální peptidy v boji s infekčními chorobami jednou mohli stát léčivy nové generace. V tomto článku jsou shrnuty jednotlivé kroky výše zmíně‑ ného procesu a představeny některé modely vzniku membránových pórů. Dále jsou nastíněny způsoby, kterými se mikroorganismy proti působení antimikrobiálních peptidů brání.
Antimicrobial peptides are an important part of nonspecific immunity of almost all living organisms; they contribute significantly to their defence against pathogenic microorganisms. A range of physical and chemical properties important for the effect of these compounds (cationicity, hydrophobicity, amphipathicity) have been described during decades of their study, and various models of their mechanism of action have been proposed. First stage of the action is the electrostatic interaction of cationic peptides with negatively charged surface of the bacterial cell. Peptide molecules adopt their amphipathic secondary structure in the environment of bacterial membranes, which enables hydrophobic amino acid side chains to immerse inside the phospholipid bilayer of the membrane. It leads to membrane disruption, or alternatively to transmembrane pore formation. Such a loss of biological functions of bacterial membrane results in destruction of the entire cell. The effect is quick and completely different from the effect of commonly used antibiotics. Due to this effect, it is expec‑ ted that antimicrobial peptides could become the antimicrobials of new generation. In this article, the whole process as described above is discussed in detail and some membrane pore forming models are presented. In addition, modes of microbial resistance towards antimicrobial peptides are also summarised.
BACKGROUND: The study used a set of 407 human extraintestinal pathogenic E. coli strains (ExPEC) isolated from (1) skin and soft tissue infections, (2) respiratory infections, (3) intra-abdominal infections, and (4) genital smears. The set was tested for bacteriocin production, for prevalence of bacteriocin and virulence determinants, and for phylogenetic typing. Results obtained from the group of ExPEC strains were compared to data from our previously published analyses of 1283 fecal commensal E. coli strains. RESULTS: The frequency of bacteriocinogeny was significantly higher in the set of ExPEC strains (63.1 %), compared to fecal E. coli (54.2 %; p < 0.01). Microcin producers and microcin determinants dominated in ExPEC strains, while colicin producers and colicin determinants were more frequent in fecal E. coli (p < 0.01). Higher production of microcin M and lower production of microcin B17, colicin Ib, and Js was detected in the set of ExPEC strains. ExPEC strains had a significantly higher prevalence of phylogenetic group B2 (52.6 %) compared to fecal E. coli strains (38.3 %; p < 0.01). CONCLUSIONS: Human ExPEC strains were shown to differ from human fecal strains in a number of parameters including bacteriocin production, prevalence of several bacteriocin and virulence determinants, and prevalence of phylogenetic groups. Differences in these parameters were also identified within subgroups of ExPEC strains of diverse origin. While some microcin determinants (mM, mH47) were associated with virulent strains, other bacteriocin types (mB17, Ib, and Js) were associated with fecal flora.
- MeSH
- Genes, Bacterial MeSH
- Bacteriocins classification genetics metabolism MeSH
- Child MeSH
- DNA, Bacterial genetics MeSH
- Adult MeSH
- Respiratory System microbiology MeSH
- Extraintestinal Pathogenic Escherichia coli genetics isolation & purification metabolism pathogenicity MeSH
- Virulence Factors genetics metabolism MeSH
- Feces microbiology MeSH
- Phylogeny * MeSH
- Gastrointestinal Tract microbiology MeSH
- Reproductive Tract Infections microbiology MeSH
- Escherichia coli Infections microbiology MeSH
- Infant MeSH
- Colicins metabolism MeSH
- Skin microbiology MeSH
- Middle Aged MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Child, Preschool MeSH
- Prevalence * MeSH
- Aged, 80 and over MeSH
- Aged MeSH
- Check Tag
- Child MeSH
- Adult MeSH
- Infant MeSH
- Middle Aged MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Male MeSH
- Child, Preschool MeSH
- Aged, 80 and over MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Czech Republic MeSH
BACKGROUND: A set of 1181 E. coli strains of human fecal origin isolated in the South Moravia region of the Czech Republic was collected during the years 2007-2010. Altogether, 17 virulence determinants and 31 bacteriocin-encoding genes were tested in each of them. RESULTS: The occurrence of bacteriocin-encoding genes was found to be positively correlated with the occurrence of E. coli virulence factors. Based on the presence of virulence factors and their combinations, E. coli strains were classified as non-pathogenic E. coli (n = 399), diarrhea-associated E. coli (n = 179) and ExPEC strains (n = 603). Non-pathogenic and diarrhea-associated E. coli strains had a low frequency of bacteriocinogeny (32.6% and 36.9%, respectively). ExPEC strains encoding S-fimbriae (sfa), P-fimbriae (pap) and having genes for aerobactin biosynthesis (aer, iucC), α-hemolysis (α-hly) and cytotoxic necrosis factor (cnf1) were often bacteriocinogenic (73.8%), had a high prevalence of bacteriocin multi-producers and showed a higher frequency of genes encoding microcins H47, M, V, B17 and colicins E1, Ia and S4. CONCLUSIONS: The occurrence of bacteriocin-encoding genes and ExPEC virulence determinants correlate positively in E. coli strains of human fecal origin. Bacteriocin synthesis appears to modulate the ability of E. coli strains to reside in the human intestine and/or the virulence of the corresponding strains.
- MeSH
- Genes, Bacterial * MeSH
- Bacteriocins genetics MeSH
- DNA, Bacterial chemistry genetics MeSH
- Escherichia coli genetics isolation & purification MeSH
- Virulence Factors genetics MeSH
- Feces microbiology MeSH
- Genotype MeSH
- Humans MeSH
- Molecular Sequence Data MeSH
- Sequence Analysis, DNA MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Czech Republic MeSH
Extraintestinal Escherichia coli infections are associated with extraintestinal pathogenic E. coli (ExPEC) strains. A total of 114 E. coli isolates were characterized regarding their antimicrobial resistance in a prospective study of 319 broilers from 12 slaughterhouses in the Czech Republic, a European Union member, during 2008. PCR-based assays to define ExPEC-associated traits were performed in resistant strains. Consumption of antimicrobial drugs by poultry in the Czech Republic was also analyzed. Antibiotic resistance was detected in 82% of isolates. Resistance to nalidixic acid and ciprofloxacin was predominant. Plasmid-mediated quinolone resistance genes, qnrB19 and qnrS1, were detected in 1 and 3 of 93 resistant isolates, respectively. Twenty-three percent of resistant isolates were considered as ExPEC. In total, 972 kg of flumequine, enrofloxacin, and difloxacin were used in poultry in the Czech Republic during 2008. High prevalence of broilers with ciprofloxacin-resistant E. coli isolates was linked to consumption of quinolones in poultry. Broilers may comprise an important vehicle for community-wide dissemination of fluoroquinolone-resistant E. coli and ExPEC. Withdrawal of fluoroquinolones from use in chicken production should be seriously considered in the Czech Republic and the European Union as well.
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Anti-Infective Agents pharmacology MeSH
- Quinolones pharmacology MeSH
- Escherichia coli drug effects isolation & purification MeSH
- Virulence Factors genetics MeSH
- Phylogeny MeSH
- Escherichia coli Infections epidemiology microbiology veterinary MeSH
- Chickens * MeSH
- Meat microbiology MeSH
- Drug Resistance, Multiple, Bacterial MeSH
- Prospective Studies MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Czech Republic MeSH
