Identification of β-Lactamase-Encoding (<i>bla</i>) Genes in Phenotypically β-Lactam-Resistant <i>Escherichia coli</i> Isolated from Young Calves in Belgium

Guérin, Virginie; Thiry, Damien; Lucas, Pierrick; Blanchard, Yannick; Cawez, Frédéric; Mercuri, Paola Sandra; Galleni, Moreno; Saulmont, Marc; Mainil, Jacques

doi:10.1089/mdr.2020.0472

Cited by 6 publications

(5 citation statements)

References 33 publications

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“…In recent years, with the emergence of plasmid-mediated ampC s (such as bla CMY ), which are genes encoding for enzymes that confer resistance to a broader range of beta-lactam antimicrobials, ampC hyperproduction is no longer the dominant mechanism of resistance in E. coli ( Findlay et al, 2020 ). However, recently it has regained interest in the scientific community as it has been identified in livestock and humans in the UK ( Alzayn et al, 2020 ), in Netherlands ( Ceccarelli et al, 2019 ), and in Belgium ( Guerin et al, 2021 ). In several of these studies, the ampC hyperproduction mechanisms is due to the mutation −42 (C > T) and seems associated with the ST88.…”

Section: Discussionmentioning

confidence: 99%

Clonal and plasmidic dissemination of critical antimicrobial resistance genes through clinically relevant ExPEC and APEC-like lineages (ST) in the dairy cattle population of Québec, Canada

de Lagarde,

Fairbrother,

Archambault

et al. 2024

Front. Microbiol.

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Antimicrobial resistance can be effectively limited by improving the judicious use of antimicrobials in food production. However, its effect on the spread of AMR genes in animal populations is not well described. In the province of Québec, Canada, a new legislation implemented in 2019 has led to an unprecedented reduction in the use of critical antimicrobials in dairy production. We aimed to investigate the potential link between ESBL/AmpC E. coli isolated before and after legislation and to determine the presence of plasmids carrying genes responsible for critical AMR. We collected fecal samples from calves, cows, and manure pit from 87 Québec dairy farms approximately 2 years before and 2 years after the legislation came into effect. The whole genomes of 183 presumptive ESBL/AmpC E. coli isolated after cefotaxime enrichment were sequenced. Their phylogenetic characteristics (MLST, serogroup, cgMLST) and the presence of virulence and resistance genes and replicons were examined. A maximum likelihood phylogenetic tree was constructed based on single nucleotide polymorphism (SNPs). We identified 10 clonal lineages (same cgMLST) and 7 clones (SNPs ≤ 52). Isolates belonging to these clones could be found on different farms before and after the legislation, strongly suggesting a clonal spread of AMR genes in the population during this 4-year period. All isolates were multidrug resistant (MDR), with clone 2 being notable for the presence of macrolide, fluoroquinolone, and third-generation cephalosporin resistance genes. We also identified clinically relevant ExPEC (ST10) and APEC-like lineages (ST117, ST58, ST88) associated with the presence of ExPEC and APEC virulence genes, respectively. Our data also suggests the presence of one epidemic plasmid belonging to the IncY incompatibility group and carrying qnrs1 and blaCTX–M–15. We demonstrated that AMR genes spread through farms and can persist over a 4-year period in the dairy cattle population through both plasmids and E. coli clones, despite the restriction of critical antimicrobial use. MDR ExPEC and APEC-like STs are present in the normal microbiota of cattle (more frequently in calves). These data increase our knowledge on gene dissemination dynamics and highlight the fact that biosecurity measures should be enhanced in this industry to limit such dissemination.

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Section: Discussionmentioning

confidence: 99%

Clonal and plasmidic dissemination of critical antimicrobial resistance genes through clinically relevant ExPEC and APEC-like lineages (ST) in the dairy cattle population of Québec, Canada

de Lagarde,

Fairbrother,

Archambault

et al. 2024

Front. Microbiol.

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“…The whole procedure, including the analysis of the results, followed the European Committee on Antimicrobial Susceptibility Testing and "Comité de l'antibiogramme de la Société Française de Microbiologie" (EUCAST/CASFM) guidelines. The resistance profiles were classified in the five resistance categories as described earlier [2]: narrow-spectrum beta lactamases (NSBL: resistant to AMX and variable to AMC), extended spectrum beta lactamases (ESBL: resistant to AMX, XNL, CFQ and CTX), cephalosporinases (AmpC: resistant to AMX, AMC, XNL, CFQ, CTX, CTC and FOX), cephalosporinase-like (AmpC-like: resistant to AMX, AMC and FOX), and carbapenemases (CP: resistant to all eight beta-lactams).…”

Section: Disk Diffusion Assaymentioning

confidence: 99%

“…In Gram-negative bacteria, acquired resistance to β-lactam antibiotics is most frequently mediated by the production of plasmid-encoded β-lactamase (BLA) enzymes hydrolysing the β-lactam ring [1]. There are five categories depending on their phenotypic β-lactam inactivation spectrums: "Narrow-Spectrum-β-lactamases" (NSBL), "Extended-Spectrum-β-lactamases" (ESBL), "Cephalosporinases" (AmpC), "NSBL with cefoxitin resistance" (AmpC-like), and "Carbapenemases" (CP) [2,3]. Amongst the Gram-negative bacteria, the first BLA enzyme was actually described in Escherichia coli [4], naturally present in the intestinal microbiota of humans and different animal species [5].…”

Section: Introductionmentioning

confidence: 99%

“…Antibiotics, including β-lactams, are, therefore, widely used in human and veterinary medicine, contributing to the development of resistances [7,8]. Since the 1960s, E. coli has acquired several genes encoding many families of NSBL, ESBL, AmpC, and CP [2,3]. Moreover, these different BLA families can comprise several dozens of variants [9].…”

Section: Introductionmentioning

confidence: 99%

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Seven-Year Evolution of β-Lactam Resistance Phenotypes in Escherichia coli Isolated from Young Diarrheic and Septicaemic Calves in Belgium

Guérin

Farchi

Thiry

et al. 2022

Veterinary Sciences

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Antimicrobial resistance is a major worldwide hazard. Therefore, the World Health Organization has proposed a classification of antimicrobials with respect to their importance for human medicine and advised some restriction of their use in veterinary medicine. In Belgium, this regulation has been implemented by a Royal Decree (RD) in 2016, which prohibits carbapenem use and enforces strict restrictions on the use of third- and fourth-generation cephalosporins (3 GC and 4 GC) for food-producing animals. Acquired resistance to β-lactam antibiotics is most frequently mediated by the production of β-lactamases in Gram-negative bacteria. This study follows the resistance to β-lactam antibiotics in Escherichia coli isolated from young diarrheic or septicaemic calves in Belgium over seven calving seasons in order to measure the impact of the RD. Phenotypic resistance to eight β-lactams was assessed by disk diffusion assay and isolates were assigned to four resistance profiles: narrow-spectrum β-lactamases (NSBL); extended-spectrum β-lactamases (ESBL); cephalosporinases (AmpC); and cephalosporinase-like, NSBL with cefoxitin resistance (AmpC-like). No carbapenemase-mediated resistance was detected. Different resistance rates were observed for each profile over the calving seasons. Following the RD, the number of susceptibility tests has increased, the resistance rate to 3 GC/4 GC has markedly decreased, while the observed resistance profiles have changed, with an increase in NSBL profiles in particular.

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“…The “RUBLA” project which studied the distribution of bla genes in bovine E. coli isolates in Wallonia, Belgium highlighted that the bla CMY-2 coding for the cephalosporinase CMY-2 is characterized by the broadest geographic spread (20). In addition, Enterobacteriaceae strains expressing this β-lactamase were isolated from animal (22) and human sources (23).…”

Section: Introductionmentioning

confidence: 99%

Development of nanobodies as theranostic agents against CMY-2-like class C β-lactamases

Cawez

Mercuri

Yanez

et al. 2022

Preprint

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Soluble single-domain fragments derived from the unique variable region of camelid heavy-chain antibodies (VHHs) against enzymes may behave as potent inhibitors. The immunization of alpacas with the CMY-2 β-lactamase led to the isolation of three VHHs that specifically recognized and inhibited CMY-2. The structure of the complex VHH cAbCMY-2(254)/CMY-2 was determined by X-ray crystallography. We showed that the epitope is close to the active site and that the CDR3 of the VHH protrudes in the catalytic site. The β-lactamase inhibition was found to follow a mixed profile with a predominant non-competitive component. The three isolated VHHs recognized overlapping epitopes since they behaved as competitive binder. Our study identified a binding site that can be targeted by a new class of β-lactamase's inhibitors designed with the help of a peptidomimetic approach. Furthermore, the use of mono or bivalent VHH and rabbit polyclonal anti-CMY-2 antibodies enable the development of the first generation of ELISA test for the detection of CMY-2 produced by resistant bacteria.

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Identification of β-Lactamase-Encoding (bla) Genes in Phenotypically β-Lactam-Resistant Escherichia coli Isolated from Young Calves in Belgium

Cited by 6 publications

References 33 publications

Clonal and plasmidic dissemination of critical antimicrobial resistance genes through clinically relevant ExPEC and APEC-like lineages (ST) in the dairy cattle population of Québec, Canada

Clonal and plasmidic dissemination of critical antimicrobial resistance genes through clinically relevant ExPEC and APEC-like lineages (ST) in the dairy cattle population of Québec, Canada

Seven-Year Evolution of β-Lactam Resistance Phenotypes in Escherichia coli Isolated from Young Diarrheic and Septicaemic Calves in Belgium

Development of nanobodies as theranostic agents against CMY-2-like class C β-lactamases

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