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      Genetic Characterization of AmpC and Extended-Spectrum Beta-Lactamase Phenotypes in Escherichia coli and Salmonella From Alberta Broiler Chickens

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          Abstract

          Horizontal gene transfer is an important mechanism which facilitates bacterial populations in overcoming antimicrobial treatment. In this study, a total of 120 Escherichia coli and 62 Salmonella enterica subsp. enterica isolates were isolated from broiler chicken farms in Alberta. Fourteen serovars were identified among Salmonella isolates. Thirty one percent of E. coli isolates (37/120) were multiclass drug resistant (resistant to ≥ 3 drug classes), while only about 16% of Salmonella isolates (10/62) were multiclass drug resistant. Among those, eight E. coli isolates had an AmpC-type phenotype, and one Salmonella isolate had an extended-spectrum beta-lactamase (ESBL)-type beta-lactamase phenotype. We identified both AmpC-type ( bla CMY-2) and ESBL-type ( bla TEM) genes in both E. coli and Salmonella isolates. Plasmids from eight of nine E. coli and Salmonella isolates were transferred to recipient strain E. coli J53 through conjugation. Transferable plasmids in the eight E. coli and Salmonella isolates were also transferred into a lab-made sodium azide-resistant Salmonella recipient through conjugation. The class 1 integrase gene, int1, was detected on plasmids from two E. coli isolates. Further investigation of class 1 integron cassette regions revealed the presence of an aadA gene encoding streptomycin 3’’-adenylyltransferase, an aadA1a/ aadA2 gene encoding aminoglycoside 3’’-O-adenyltransferase, and a putative adenylyltransferase gene. This study provides some insight into potential horizontal gene transfer events of antimicrobial resistance genes between E. coli and Salmonella in broiler chicken production.

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          Resistance plasmid families in Enterobacteriaceae.

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            The Clermont Escherichia coli phylo-typing method revisited: improvement of specificity and detection of new phylo-groups.

            There is extensive genetic substructure within the species Escherichia coli. In 2000 a simple triplex PCR method was described by Clermont and colleagues that enables an E. coli isolate to be assigned to one of the phylo-groups A, B1, B2 or D. The growing body of multi-locus sequence data and genome data for E. coli has refined our understanding of E. coli's phylo-group structure and eight phylo-groups are now recognized: seven (A, B1, B2, C, D, E, F) belong to E. coli sensu stricto, whereas the eighth is the Escherichia cryptic clade I. Here a new PCR-based method is developed that enables an E. coli isolate to be assigned to one of the eight phylo-groups and which allows isolates that are members of the other cryptic clades (II to V) of Escherichia to be identified. The development of the method is described and the method is validated. Over 95% of E. coli isolates can be correctly assigned to a phylo-group. Two collections of human faecal isolates were screened using the new phylo-group assignment method demonstrating that about 13% of E. coli isolates belong to the newly described phylo-groups C, E, F and clade I. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.
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              CTX-M Enzymes: Origin and Diffusion

              CTX-M β-lactamases are considered a paradigm in the evolution of a resistance mechanism. Incorporation of different chromosomal bla CTX-M related genes from different species of Kluyvera has derived in different CTX-M clusters. In silico analyses have shown that this event has occurred at least nine times; in CTX-M-1 cluster (3), CTX-M-2 and CTX-M-9 clusters (2 each), and CTX-M-8 and CTX-M-25 clusters (1 each). This has been mainly produced by the participation of genetic mobilization units such as insertion sequences (ISEcp1 or ISCR1) and the later incorporation in hierarchical structures associated with multifaceted genetic structures including complex class 1 integrons and transposons. The capture of these bla CTX-M genes from the environment by highly mobilizable structures could have been a random event. Moreover, after incorporation within these structures, β-lactam selective force such as that exerted by cefotaxime and ceftazidime has fueled mutational events underscoring diversification of different clusters. Nevertheless, more variants of CTX-M enzymes, including those not inhibited by β-lactamase inhibitors such as clavulanic acid (IR-CTX-M variants), only obtained under in in vitro experiments, are still waiting to emerge in the clinical setting. Penetration and the later global spread of CTX-M producing organisms have been produced with the participation of the so-called “epidemic resistance plasmids” often carried in multi-drug resistant and virulent high-risk clones. All these facts but also the incorporation and co-selection of emerging resistance determinants within CTX-M producing bacteria, such as those encoding carbapenemases, depict the currently complex pandemic scenario of multi-drug resistant isolates.
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                Author and article information

                Contributors
                Journal
                Front Cell Infect Microbiol
                Front Cell Infect Microbiol
                Front. Cell. Infect. Microbiol.
                Frontiers in Cellular and Infection Microbiology
                Frontiers Media S.A.
                2235-2988
                12 March 2021
                2021
                : 11
                : 622195
                Affiliations
                [1] 1 Department of Ecosystem and Public Health, Faculty of Veterinary Medicine, University of Calgary , Calgary, AB, Canada
                [2] 2 Animal Policy and Epidemiology Section, Animal Health Branch, Animal Health and Assurance Division, Alberta Agriculture and Forestry , Edmonton, AB, Canada
                [3] 3 Western College of Veterinary Medicine, University of Saskatchewan , Saskatoon, SK, Canada
                [4] 4 Center for Foodborne, Environmental and Zoonotic Infectious Diseases, Public Health Agency of Canada , Guelph, ON, Canada
                Author notes

                Edited by: Fangkun Wang, Shandong Agricultural University, China

                Reviewed by: Mohamed Salah Abbassi, Tunis El Manar University, Tunisia; Xiaoping Liao, South China Agricultural University, China; Toshiyuki Murase, Tottori University, Japan

                *Correspondence: Karen Liljebjelke, kliljebj@ 123456ucalgary.ca ; Tam Tran, ttran332@ 123456uwo.ca

                This article was submitted to Molecular Bacterial Pathogenesis, a section of the journal Frontiers in Cellular and Infection Microbiology

                Article
                10.3389/fcimb.2021.622195
                7994595
                1c77ebac-4d8d-4d06-a4dc-d9091ad61d1c
                Copyright © 2021 Tran, Checkley, Caffrey, Mainali, Gow, Agunos and Liljebjelke

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 27 October 2020
                : 18 February 2021
                Page count
                Figures: 1, Tables: 7, Equations: 0, References: 53, Pages: 12, Words: 6502
                Funding
                Funded by: Alberta Agriculture and Forestry 10.13039/100012236
                Categories
                Cellular and Infection Microbiology
                Original Research

                Infectious disease & Microbiology
                escherichia coli,salmonella,blacmy-2,blatem,antimicrobial resistance genes

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