Journal: Frontiers in Microbiology

Article Title: Occurrence of the mcr-1 Colistin Resistance Gene and other Clinically Relevant Antibiotic Resistance Genes in Microbial Populations at Different Municipal Wastewater Treatment Plants in Germany

doi: 10.3389/fmicb.2017.01282

Figure Lengend Snippet: Abundances of taxonomical marker genes per 100 ng of total extracted DNA from all 7 WWTPs. (A) A. baumannii was detected by secE gene abundance, (B) K. pneumoniae was detected by gltA gene abundance, and (C) E. coli was detected by yccT gene abundance. Displayed are the median values as well as the quantils [ p = 0.25 (dark), p = 0.75 (bright)] and the standard deviations ( n = 14). Significance of reduction is assessed by student's t- test with p ≤ 0.05 and are indicated by asterisk.

Article Snippet: More specifically, the Escherichia coli NRZ-14408 reference strain is carrying the mcr-1 colistin resistance gene and was kindly provided from National Reference Center in Bochum, Germany.

Techniques: Marker

Journal: Frontiers in Microbiology

Article Title: Occurrence of the mcr-1 Colistin Resistance Gene and other Clinically Relevant Antibiotic Resistance Genes in Microbial Populations at Different Municipal Wastewater Treatment Plants in Germany

doi: 10.3389/fmicb.2017.01282

Figure Lengend Snippet: Abundances of taxonomical marker genes per 16S rRNA gene copy number derived from total extracted DNA of all seven WWTPs. (A) A. baumannii was detected by secE gene abundance, (B) K. pneumoniae was detected by gltA gene abundance, and (C) E. coli was detected by yccT gene abundance. Displayed are the median values as well as the quantils [ p = 0.25 (dark), p = 0.75 (bright)] and the standard deviations ( n = 14). Student's t -test values were all >0.05.

Article Snippet: More specifically, the Escherichia coli NRZ-14408 reference strain is carrying the mcr-1 colistin resistance gene and was kindly provided from National Reference Center in Bochum, Germany.

Techniques: Marker, Derivative Assay

Journal: Glycobiology

Article Title: Elucidation of the O-antigen structure of Escherichia coli O93 and characterization of its biosynthetic genes

doi: 10.1093/glycob/cwac069

Figure Lengend Snippet: SDS-PAGE analysis of LPS from different E. coli strains. Lane 1: LPS from E. coli CWG303 Δ waaG , Lane 2: LPS from E. coli W3110, and Lane 3: LPS from E. coli O93. A 14% Tricine SDS-PAGE gel was used to separate LPS samples, which subsequently were stained with AgNO 3 . A pre-stained protein marker (Thermo Fisher Scientific) was utilized for calibration of molecular weights.

Article Snippet: Escherichia coli reference strains of O93 ( E. coli O93:−:H16) was obtained from SSI Diagnostica A/S (Hillerød, Denmark).

Techniques: SDS Page, Staining, Marker

Journal: Glycobiology

Article Title: Elucidation of the O-antigen structure of Escherichia coli O93 and characterization of its biosynthetic genes

doi: 10.1093/glycob/cwac069

Figure Lengend Snippet: The O-antigen gene cluster between galF and hisI in E. coli O93 ( Iguchi et al. 2015 ; DebRoy et al. 2016 ; Liu et al. 2020 ).

Article Snippet: Escherichia coli reference strains of O93 ( E. coli O93:−:H16) was obtained from SSI Diagnostica A/S (Hillerød, Denmark).

Techniques:

Journal: Glycobiology

Article Title: Elucidation of the O-antigen structure of Escherichia coli O93 and characterization of its biosynthetic genes

doi: 10.1093/glycob/cwac069

Figure Lengend Snippet: The anomeric region of O -deacetylated E. coli O93 O-antigen polysaccharide (PS–OH) from a 1 H NMR spectrum (a) and a 1 H, 1 H-NOESY spectrum with a mixing time of 150 ms at 700 MHz (b). Transglycosidic cross-peaks are annotated by red color.

Article Snippet: Escherichia coli reference strains of O93 ( E. coli O93:−:H16) was obtained from SSI Diagnostica A/S (Hillerød, Denmark).

Techniques:

Journal: Glycobiology

Article Title: Elucidation of the O-antigen structure of Escherichia coli O93 and characterization of its biosynthetic genes

doi: 10.1093/glycob/cwac069

Figure Lengend Snippet: NMR spectra of selected regions from the O-antigen polysaccharides (PS and PS–OH) of E. coli O93. (a) An overlay of 1 H, 13 C-meHSQC NMR spectra of the anomeric region from PS (black) and PS–OH (red); (b) 1 H, 13 C-meHSQC spectrum of PS–OH plotted in red color with linkage positions annotated and hydroxymethyl correlations plotted in blue color; (c, d) selected regions from the 1 H, 13 C-meHSQC spectrum of the PS sample showing the O -acetylated positions in residues C′ / C″ with hydroxymethyl correlations plotted in green color; (e, f) 1 H, 1 H-TOCSY spectrum with a mixing time of 120 ms displaying the cross-peaks from H2 resonances in residues C2′ and C2″ ; (g) selected region from the coupling-enhanced (2×) 1 H, 13 C-CT-CE-HSQC NMR spectrum resulting in 1 J C1,H1 for B′/B″ (158 Hz) and C′/C″ (160 Hz).

Article Snippet: Escherichia coli reference strains of O93 ( E. coli O93:−:H16) was obtained from SSI Diagnostica A/S (Hillerød, Denmark).

Techniques:

Journal: Glycobiology

Article Title: Elucidation of the O-antigen structure of Escherichia coli O93 and characterization of its biosynthetic genes

doi: 10.1093/glycob/cwac069

Figure Lengend Snippet: Top 3 structures using the “Determine Glycan Structure” entry in CASPER for E. coli O93 PS–OH with NMR data obtained from 1 H, 13 C-HSQC and 1 H, 1 H-TOCSY experiments and magnitudes of coupling constants for input to CASPER, viz., those from 1 J C1,H1 and 3 J H1,H2 obtained from coupled 1 H, 13 C-HSQC and 1 H NMR spectra, respectively.

Article Snippet: Escherichia coli reference strains of O93 ( E. coli O93:−:H16) was obtained from SSI Diagnostica A/S (Hillerød, Denmark).

Techniques: Glycoproteomics

Journal: Glycobiology

Article Title: Elucidation of the O-antigen structure of Escherichia coli O93 and characterization of its biosynthetic genes

doi: 10.1093/glycob/cwac069

Figure Lengend Snippet: 1 H and 13 C NMR chemical shifts of the O-antigen from E. coli O93 in which populations carrying O -acetyl groups are denoted by prime and double-prime residues and the corresponding O -deacetylated polysaccharide where sugar residues are devoid of the prime character.

Article Snippet: Escherichia coli reference strains of O93 ( E. coli O93:−:H16) was obtained from SSI Diagnostica A/S (Hillerød, Denmark).

Techniques: Residue

Journal: Glycobiology

Article Title: Elucidation of the O-antigen structure of Escherichia coli O93 and characterization of its biosynthetic genes

doi: 10.1093/glycob/cwac069

Figure Lengend Snippet: Inter-residue correlations from 1 H, 1 H-NOESY, 1D 1 H, 1 H-NOESY, and 1 H, 13 C-HMBC spectra for E. coli O93 O-antigen polysaccharide (PS) and de- O -acetylated polysaccharide (PS–OH).

Article Snippet: Escherichia coli reference strains of O93 ( E. coli O93:−:H16) was obtained from SSI Diagnostica A/S (Hillerød, Denmark).

Techniques: Residue

Journal: Glycobiology

Article Title: Elucidation of the O-antigen structure of Escherichia coli O93 and characterization of its biosynthetic genes

doi: 10.1093/glycob/cwac069

Figure Lengend Snippet: Comparison of 1 H (left) and 13 C (right) NMR chemical shifts predicted by CASPER versus assigned from NMR experiments for the O -deacetylated (PS–OH) and the 2,6-di- O -acetylated population of the polysaccharide (PS) from the E. coli O93 O-antigen.

Article Snippet: Escherichia coli reference strains of O93 ( E. coli O93:−:H16) was obtained from SSI Diagnostica A/S (Hillerød, Denmark).

Techniques: Comparison

Journal: Glycobiology

Article Title: Elucidation of the O-antigen structure of Escherichia coli O93 and characterization of its biosynthetic genes

doi: 10.1093/glycob/cwac069

Figure Lengend Snippet: (top) A “Sequence plot” summarizing the connectivities observed in a 1 H, 1 H-NOESY spectrum of E. coli O93 PS–OH (upper triangle) and a 1 H, 13 C-HMBC spectrum of E. coli O93 PS (lower triangle) where the connectivities emanate “from” the anomeric position “to” the linkage position, (middle) E. coli O93 PS repeating unit in SNFG-representation ( Neelamegham et al. 2019 ) and (bottom) standard nomenclature with residues annotated by A – D .

Article Snippet: Escherichia coli reference strains of O93 ( E. coli O93:−:H16) was obtained from SSI Diagnostica A/S (Hillerød, Denmark).

Techniques: Sequencing

Journal: Glycobiology

Article Title: Elucidation of the O-antigen structure of Escherichia coli O93 and characterization of its biosynthetic genes

doi: 10.1093/glycob/cwac069

Figure Lengend Snippet: (left) 3D representation of the trisaccharide α- d -GlcNAc-(1→2)-β- d -Man-(1→3)-β- d -Man2Ac6Ac from the E. coli O93 PS in a conformation consistent with the 1D 1 H, 1 H-NOESY spectrum (right) with an irradiation at 2.22 ppm of the methyl group of the O -acetyl group at C2 recorded with a 600 ms mixing time at 700 MHz showing inter-residue correlations. The yellow conical shape represents the proposed anisotropic shielding effect by the carbonyl group of the O -acetyl group at position C2 affecting H2 in residue B by an upfield chemical shift displacement, in comparison to the O -deacetylated polysaccharide. Primes used in the text to differentiate O -acetylated populations in the polysaccharide are not included in the figure for clarity.

Article Snippet: Escherichia coli reference strains of O93 ( E. coli O93:−:H16) was obtained from SSI Diagnostica A/S (Hillerød, Denmark).

Techniques: Irradiation, Residue, Comparison

Journal: Glycobiology

Article Title: Elucidation of the O-antigen structure of Escherichia coli O93 and characterization of its biosynthetic genes

doi: 10.1093/glycob/cwac069

Figure Lengend Snippet: Protein structure obtained from the AlphaFold Protein Structure Database of a putative glycosyltransferase ( orf5 in the O-antigen gene cluster) from E. coli O93 carrying out the last glycosylation reaction in formation of the oligosaccharyl- PP -undecaprenyl entity. The molecular model was made using UCSF Chimera ( Pettersen et al. 2004 ).

Article Snippet: Escherichia coli reference strains of O93 ( E. coli O93:−:H16) was obtained from SSI Diagnostica A/S (Hillerød, Denmark).

Techniques: Glycoproteomics

Journal: BMC Veterinary Research

Article Title: Evaluation of inhibition of F4ac positive Escherichia coli attachment with xanthine dehydrogenase, butyrophilin, lactadherin and fatty acid binding protein

doi: 10.1186/s12917-015-0528-0

Figure Lengend Snippet: Quantification of inhibitory effect of MFGM proteins against attachment of F4ac-positive E. coli to the IPEC-J2 cells. Inhibition of F4ac-positive E. coli attachment to IPEC-J2 cells with various concentrations (5, 25 or 125 μg/ml) of xanthine dehydrogenase (XD), butyrophilin (BTN), lactadherin (LAD), fatty acid binding protein (FAB); positive (rabbit anti-F4ac polyclonal antibody) and negative (rabbit anti-F6 polyclonal antibody or Lactalbumin) controls. Asterisk (*) indicates statistically significant difference between an individual dilution and all of the negative controls (untreated JPEC-J2 + F4ac-positive E. coli , Lactalbumin or anti-F6 antibodies), ( P < 0.05). Different letters (a), (b) and (c) indicate statistically significant differences ( P < 0.05) between individual dilutions of the same milk fat globule membrane protein. Error bar indicates Standard Deviation. Each column represents mean of quadruplicate measurements in a single representative experiment

Article Snippet: Enterotoxigenic F4ac-positive Escherichia coli (reference strain P97-2554B, O149:K91:F4ac + EST1 + LT + STa + STb + ) isolated from the intestine of a pig with diarrhea at the faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, QC was kindly provided by Dr. Musangu Ngeleka, Prairie Diagnostic Services, Inc., Saskatoon, SK.

Techniques: Inhibition, Binding Assay, Membrane, Standard Deviation

Journal: BMC Veterinary Research

Article Title: Evaluation of inhibition of F4ac positive Escherichia coli attachment with xanthine dehydrogenase, butyrophilin, lactadherin and fatty acid binding protein

doi: 10.1186/s12917-015-0528-0

Figure Lengend Snippet: Quantification of inhibitory effect of MFGM proteins against attachment of F4ac-positive E. coli to the pig enterocytes. Inhibition of F4ac-positive E. coli attachment to porcine enterocytes with various concentrations (5, 25 or 125 μg/ml) of xanthine dehydrogenase (XD), butyrophilin (BTN), lactadherin (LAD), fatty acid binding protein (FAB); positive (rabbit anti-F4ac polyclonal antibody) and negative (rabbit anti-F6 polyclonal antibody or Lactalbumin) controls. Asterisk (*) indicates statistically significant difference between an individual dilution and all of the negative controls (untreated enterocytes + F4ac-positive E. coli , Lactalbumin or anti-F6 antibodies), ( P < 0.05). Different letters (a), (b) and (c) indicate statistically significant differences ( P < 0.05) between individual dilutions of the same milk fat globule membrane protein. Error bar indicates Standard Deviation. Each column represents mean of quadruplicate measurements in a single representative experiment

Article Snippet: Enterotoxigenic F4ac-positive Escherichia coli (reference strain P97-2554B, O149:K91:F4ac + EST1 + LT + STa + STb + ) isolated from the intestine of a pig with diarrhea at the faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, QC was kindly provided by Dr. Musangu Ngeleka, Prairie Diagnostic Services, Inc., Saskatoon, SK.

Techniques: Inhibition, Binding Assay, Membrane, Standard Deviation

Journal: BMC Veterinary Research

Article Title: Evaluation of inhibition of F4ac positive Escherichia coli attachment with xanthine dehydrogenase, butyrophilin, lactadherin and fatty acid binding protein

doi: 10.1186/s12917-015-0528-0

Figure Lengend Snippet: Isolation of F4ac fimbriae from F4ac-positive E. coli . F4ac fimbriae isolated from F4ac-positive E. coli , subjected to 12 % SDS-PAGE, and visualized by Coomassie Blue staining (a) and detected by Western blotting using primary rabbit anti-F4-polyclonal antibodies and secondary anti-rabbit Cy5 antibodies (b). Lane 1 – (a) Molecular weight markers; (b) ECL Flex Rainbow Molecular Weight Markers; Lane 2 - Bovine serum albumin as negative control; Lane 3- isolated F4ac fimbriae

Article Snippet: Enterotoxigenic F4ac-positive Escherichia coli (reference strain P97-2554B, O149:K91:F4ac + EST1 + LT + STa + STb + ) isolated from the intestine of a pig with diarrhea at the faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, QC was kindly provided by Dr. Musangu Ngeleka, Prairie Diagnostic Services, Inc., Saskatoon, SK.

Techniques: Isolation, SDS Page, Staining, Western Blot, Molecular Weight, Negative Control