Table_1_Fluoroquinolone resistance in complicated urinary tract infections: association with the increased occurrence and diversity of Escherichia coli of clonal complex 131, together with ST1193.xlsx

IntroductionUrinary tract infections (UTIs) are on3e of the leading causes of multidrug-resistance (MDR) spread and infection-related deaths. Escherichia coli is by far the main causative agent. We conducted a prospective study on complicated urinary tract infections (cUTIs) i) to monitor the high-risk clones that could be compromising the therapeutic management and ii) to compare the cUTI etiology with uncomplicated infections (uUTIs) occurring in the same period and health area.Methods154 non-duplicated E. coli recovered from cUTIs in 2020 at the Hospital Universitario Central de Asturias (Spain) constituted the study collection.ResultsMost cUTI isolates belonged to phylogroup B2 (72.1%) and met the uropathogenic (UPEC) status (69.5%) (≥3 of chuA, fyuA, vat, and yfcV genes). MDR was exhibited by 35.7% of the isolates, similarly to data observed in the uUTI collection. A significant difference observed in cUTI was the higher level of fluoroquinolone resistance (FQR) (47.4%), where the pandemic clonal groups B2-CC131 and B2-ST1193 (CH14-64) comprised 28% of the 154 E. coli, representing 52.1% of the FQR isolates. Other prevalent FQR clones were D-ST69 (CH35-27), D-ST405 (CH37-27), and B2-ST429 (CH40-20) (three isolates each). We uncovered an increased genetic and genomic diversity of the CC131: 10 different virotypes, 8 clonotypes (CH), and 2 STs. The presence of blaCTX-M-15 was determined in 12 (7.8%) isolates (all CC131), which showed 10 different core genome (cg)STs and 2 fimH types (fimH30 and fimH602) but the same set of chromosomal mutations conferring FQR (gyrA p.S83L, gyrA p.D87N, parC p.S80I, parC p.E84V, and parE p.I529L). In addition, the plasmidome analysis revealed 10 different IncF formulae in CC131 genomes.ConclusionWe proved here that non-lactose fermenting screening, together with the detection of O25b (rfbO25b), H4 (fliCH4), and H5 (fliCH5) genes, and phylogroup and clonotyping assignation, is a reasonable approach that can be easily implemented for the surveillance of emerging high-risk clones associated with FQR spread in cUTIs, such as the uncommonly reported O25b:H4-B2-ST9126-CC131 (CH1267-30). Since E. coli CC131 and ST1193 are also involved in the community uUTIs of this health area, interventions to eradicate these MDR clones, along with surveillance for other emerging ones, are essential for antibiotic use optimization programs.</p

Table_1_Fluoroquinolone resistance in complicated urinary tract infections: association with the increased occurrence and diversity of Escherichia coli of clonal complex 131, together with ST1193.xlsx

IntroductionUrinary tract infections (UTIs) are one of the leading causes of multidrug-resistance (MDR) spread and infection-related deaths. Escherichia coli is by far the main causative agent. We conducted a prospective study on complicated urinary tract infections (cUTIs) i) to monitor the high-risk clones that could be compromising the therapeutic management and ii) to compare the cUTI etiology with uncomplicated infections (uUTIs) occurring in the same period and health area.Methods154 non-duplicated E. coli recovered from cUTIs in 2020 at the Hospital Universitario Central de Asturias (Spain) constituted the study collection.ResultsMost cUTI isolates belonged to phylogroup B2 (72.1%) and met the uropathogenic (UPEC) status (69.5%) (≥3 of chuA, fyuA, vat, and yfcV genes). MDR was exhibited by 35.7% of the isolates, similarly to data observed in the uUTI collection. A significant difference observed in cUTI was the higher level of fluoroquinolone resistance (FQR) (47.4%), where the pandemic clonal groups B2-CC131 and B2-ST1193 (CH14-64) comprised 28% of the 154 E. coli, representing 52.1% of the FQR isolates. Other prevalent FQR clones were D-ST69 (CH35-27), D-ST405 (CH37-27), and B2-ST429 (CH40-20) (three isolates each). We uncovered an increased genetic and genomic diversity of the CC131: 10 different virotypes, 8 clonotypes (CH), and 2 STs. The presence of blaCTX-M-15 was determined in 12 (7.8%) isolates (all CC131), which showed 10 different core genome (cg)STs and 2 fimH types (fimH30 and fimH602) but the same set of chromosomal mutations conferring FQR (gyrA p.S83L, gyrA p.D87N, parC p.S80I, parC p.E84V, and parE p.I529L). In addition, the plasmidome analysis revealed 10 different IncF formulae in CC131 genomes.ConclusionWe proved here that non-lactose fermenting screening, together with the detection of O25b (rfbO25b), H4 (fliCH4), and H5 (fliCH5) genes, and phylogroup and clonotyping assignation, is a reasonable approach that can be easily implemented for the surveillance of emerging high-risk clones associated with FQR spread in cUTIs, such as the uncommonly reported O25b:H4-B2-ST9126-CC131 (CH1267-30). Since E. coli CC131 and ST1193 are also involved in the community uUTIs of this health area, interventions to eradicate these MDR clones, along with surveillance for other emerging ones, are essential for antibiotic use optimization programs.</p

DataSheet_1_Fluoroquinolone resistance in complicated urinary tract infections: association with the increased occurrence and diversity of Escherichia coli of clonal complex 131, together with ST1193.docx

IntroductionUrinary tract infections (UTIs) are on3e of the leading causes of multidrug-resistance (MDR) spread and infection-related deaths. Escherichia coli is by far the main causative agent. We conducted a prospective study on complicated urinary tract infections (cUTIs) i) to monitor the high-risk clones that could be compromising the therapeutic management and ii) to compare the cUTI etiology with uncomplicated infections (uUTIs) occurring in the same period and health area.Methods154 non-duplicated E. coli recovered from cUTIs in 2020 at the Hospital Universitario Central de Asturias (Spain) constituted the study collection.ResultsMost cUTI isolates belonged to phylogroup B2 (72.1%) and met the uropathogenic (UPEC) status (69.5%) (≥3 of chuA, fyuA, vat, and yfcV genes). MDR was exhibited by 35.7% of the isolates, similarly to data observed in the uUTI collection. A significant difference observed in cUTI was the higher level of fluoroquinolone resistance (FQR) (47.4%), where the pandemic clonal groups B2-CC131 and B2-ST1193 (CH14-64) comprised 28% of the 154 E. coli, representing 52.1% of the FQR isolates. Other prevalent FQR clones were D-ST69 (CH35-27), D-ST405 (CH37-27), and B2-ST429 (CH40-20) (three isolates each). We uncovered an increased genetic and genomic diversity of the CC131: 10 different virotypes, 8 clonotypes (CH), and 2 STs. The presence of blaCTX-M-15 was determined in 12 (7.8%) isolates (all CC131), which showed 10 different core genome (cg)STs and 2 fimH types (fimH30 and fimH602) but the same set of chromosomal mutations conferring FQR (gyrA p.S83L, gyrA p.D87N, parC p.S80I, parC p.E84V, and parE p.I529L). In addition, the plasmidome analysis revealed 10 different IncF formulae in CC131 genomes.ConclusionWe proved here that non-lactose fermenting screening, together with the detection of O25b (rfbO25b), H4 (fliCH4), and H5 (fliCH5) genes, and phylogroup and clonotyping assignation, is a reasonable approach that can be easily implemented for the surveillance of emerging high-risk clones associated with FQR spread in cUTIs, such as the uncommonly reported O25b:H4-B2-ST9126-CC131 (CH1267-30). Since E. coli CC131 and ST1193 are also involved in the community uUTIs of this health area, interventions to eradicate these MDR clones, along with surveillance for other emerging ones, are essential for antibiotic use optimization programs.</p

DataSheet_1_Fluoroquinolone resistance in complicated urinary tract infections: association with the increased occurrence and diversity of Escherichia coli of clonal complex 131, together with ST1193.docx

IntroductionUrinary tract infections (UTIs) are one of the leading causes of multidrug-resistance (MDR) spread and infection-related deaths. Escherichia coli is by far the main causative agent. We conducted a prospective study on complicated urinary tract infections (cUTIs) i) to monitor the high-risk clones that could be compromising the therapeutic management and ii) to compare the cUTI etiology with uncomplicated infections (uUTIs) occurring in the same period and health area.Methods154 non-duplicated E. coli recovered from cUTIs in 2020 at the Hospital Universitario Central de Asturias (Spain) constituted the study collection.ResultsMost cUTI isolates belonged to phylogroup B2 (72.1%) and met the uropathogenic (UPEC) status (69.5%) (≥3 of chuA, fyuA, vat, and yfcV genes). MDR was exhibited by 35.7% of the isolates, similarly to data observed in the uUTI collection. A significant difference observed in cUTI was the higher level of fluoroquinolone resistance (FQR) (47.4%), where the pandemic clonal groups B2-CC131 and B2-ST1193 (CH14-64) comprised 28% of the 154 E. coli, representing 52.1% of the FQR isolates. Other prevalent FQR clones were D-ST69 (CH35-27), D-ST405 (CH37-27), and B2-ST429 (CH40-20) (three isolates each). We uncovered an increased genetic and genomic diversity of the CC131: 10 different virotypes, 8 clonotypes (CH), and 2 STs. The presence of blaCTX-M-15 was determined in 12 (7.8%) isolates (all CC131), which showed 10 different core genome (cg)STs and 2 fimH types (fimH30 and fimH602) but the same set of chromosomal mutations conferring FQR (gyrA p.S83L, gyrA p.D87N, parC p.S80I, parC p.E84V, and parE p.I529L). In addition, the plasmidome analysis revealed 10 different IncF formulae in CC131 genomes.ConclusionWe proved here that non-lactose fermenting screening, together with the detection of O25b (rfbO25b), H4 (fliCH4), and H5 (fliCH5) genes, and phylogroup and clonotyping assignation, is a reasonable approach that can be easily implemented for the surveillance of emerging high-risk clones associated with FQR spread in cUTIs, such as the uncommonly reported O25b:H4-B2-ST9126-CC131 (CH1267-30). Since E. coli CC131 and ST1193 are also involved in the community uUTIs of this health area, interventions to eradicate these MDR clones, along with surveillance for other emerging ones, are essential for antibiotic use optimization programs.</p

Corrosion Resistance Evaluation of HVOF Produced Hydroxyapatite and TiO2-hydroxyapatite Coatings in Hanks' Solution

<div><p>The electrochemical behavior of HVOF produced hydroxyapatite (HA) and 80HA-20TiO2 coatings were investigated using electrochemical techniques in natural aerated Hanks' solution in the presence and absence of bovine serum albumin (BSA) for 30 days. All samples presented open circuit potential oscillations, which were associated to the porous nature of the coating that allows the electrolyte reaches the substrate causing activation - passivation at the bottom of the pores. The polarization studies indicated that the 80HA-20TiO2 coating was the only one that showed a narrow potential passive region from around -0.4 V to 0 V in the presence and absence of BSA, indicating the beneficial influence of the addition of TiO2 to the HA coating stability. Our results indicated that BSA in Hanks' solution diminishes the stability of the metallic oxide layer present on the Ti-based alloy accelerating the degrading of hydroxyapatites coatings / substrate interface due to its chelating ability.</p></div

Confocal microscopy images of <i>E</i>. <i>coli</i> (A–G, a–g), <i>Salmonella enterica</i> (H, h) and <i>Enterobacter cloacae</i> (I, i) fluorescent labeling tests.

<p>Each group of 4 images correspond to a different chimeric biosensor fluorescent protein. Images from “A” to “I” correspond to confocal microscopy, showing the fluorescent labelled cells, and images from “a” to “i” are the same confocal images but fused with an optical image generating a single photo (merged snapshot for testing that fluorescent spots actually correspond to labelled cells, and not to background fluorescence). The symbol “✽” means that the sample has been filtered in order to get rid of unbound chimera protein. A-B, images of <i>E</i>. <i>coli</i> labelled with GFP-hadrurin fluorescent chimera protein, where labeling is lost after a filtering step. C-D, images of <i>E</i>. <i>coli</i> labelled with GFP-pb5 fluorescent chimera protein, where labeling is also lost after a filtering step. Only the fluorescent biosensor chimera protein GFP-colS4 (E,F) is able to maintain its strong binding to the <i>E</i>. <i>coli</i> surface after the filtration step. Also, the other two bacterial species (negative controls) are not labelled at all (H, I). Images G and g are negative controls for <i>E</i>. <i>coli</i>, where no fluorescent biosensor chimera protein has been added, as a method to test that this bacterial cells do not show autofluorescence under these conditions.</p

Functional blocks of the measurement system at the portable <i>E</i>. <i>coli</i> Analyzer device.

<p>Functional blocks of the measurement system at the portable <i>E</i>. <i>coli</i> Analyzer device.</p

MUSCLE multiple sequence alignment of the OmpW proteins from <i>E</i>. <i>coli</i>, <i>S</i>. <i>enterica</i> var. <i>arizonae</i> and <i>E</i>. <i>cloacae</i>, highlighting in yellow the region encompasing the the Asp¹¹⁶, His¹¹⁷ and Glu¹²⁰ amino acids of OmpW (in blue letters: D, H and E), which are freely exposed to the extracellular space and act as binding motifs for colicin S4 bacteriocin (as well as for GFP-colS4 chimera protein in this biosensor).

<p>As it is shown, the other two enterobacteria species do not have the conserved motif DH—E as in <i>E</i>. <i>coli</i>. Accesion numbers: SAI89619 (<i>E</i>. <i>cloacae</i>), BAA14788 (<i>E</i>. <i>coli</i>), OSE54138 (<i>S</i>. <i>enterica</i>).</p

Development of a biosensor protein bullet as a fluorescent method for fast detection of <i>Escherichia coli</i> in drinking water - Fig 2

<p>A: Hypothetical spatial structure of the fluorescent biosensor chimera protein GFP-hadrurin. B: Diagram showing the attachement process of the fluorescent biosensor chimera protein to the membrane of <i>E</i>. <i>coli</i>. PMT: photomultiplier (part of the <i>E</i>. <i>coli</i> Analyzer device in charge of detecting and amplifying the fluorescence signal produced by the GFP domain of each biosensor chimera protein, after receiving the 395 nm excitation light from the LED source).</p

Values of urinary biomarkers in patients categorized according to the absence or presence of Acute-on-chronic liver failure (ACLF).

<p>Values are expressed as median and IQR.</p><p>ACLF, Acute-on-chronic liver failure</p><p>Values of urinary biomarkers in patients categorized according to the absence or presence of Acute-on-chronic liver failure (ACLF).</p