Mobility of β-Lactam Resistance Under Bacterial Co-infection and Ampicillin Treatment in a Mouse Model

Laskey, Alexander; Ottenbrite, Marie; Devenish, John; Kang, Mingsong; Savic, Mirjana; Nadin-Davis, Susan A.; Chmara, John; Lin, Min; Robertson, James A.; Bessonov, Kyrylo; Gurnik, Simone; Liu, Kira; Nash, John H. E.; Scott, Andrew; Topp, Edward; Guan, Jiewen

doi:10.3389/fmicb.2020.01591

Cited by 8 publications

(26 citation statements)

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“…IncI2 is a conjugative plasmid encoding a bla CMY-2 gene and IncY is a mobilizable plasmid encoding a bla TEM-1B gene. In vitro and in vivo transfer of the IncI2 but not IncY plasmid was detected using E. coli O80:H26 as donor and E. coli O16:H48 as recipient in the previous study [ 21 ].…”

Section: Methodsmentioning

confidence: 98%

“… E. coli O80:H26 (EC-107) is a multi-antibiotic-resistant strain isolated from a chicken farm in Ontario, Canada. E. coli O80:H26 carries five plasmids: IncI2, IncY, IncFII, ColRNAI and a plasmid with no detectable Inc type [ 21 ]. IncI2 is a conjugative plasmid encoding a bla CMY-2 gene and IncY is a mobilizable plasmid encoding a bla TEM-1B gene.…”

Section: Methodsmentioning

confidence: 99%

“…Putative transconjugant bacteria were whole-genome sequenced and sequence data were analysed using the MOB-suite software tool v2.1.0 [23,24]. Representative putative transconjugant colonies isolated from selective agar plates (up to five colonies per time point) were subjected to genomic DNA extraction as described by Laskey et al [21]. Whole-genome sequencing was performed using an Illumina MiSeq system and/or an Oxford Nanopore MinION sequencer (Oxford Nanopore, Cambridge, MA, USA) at the National Microbiology Laboratory (Guelph, ON, Canada).…”

Section: Whole Genome Sequencingmentioning

confidence: 99%

“…In vitro conjugation between E. coli O80:H26 (donor) and S. Heidelberg (recipient) was assessed in 10 −1 × LB broth as described by Laskey et al [21]. Enumeration of the donor, recipient and transconjugant bacteria was performed using Chromocult agar (EMD Millipore, Toronto, ON, Canada) supplemented with 4 µg ml −1 cefotaxime (CHR-F), XLT4 agar (Difco) with 50 ug ml −1 rifampicin (XLT4-R) and XLT4 agar with 4 µg ml −1 cefotaxime and 50 µg ml −1 rifampicin (XLT4-FR), respectively.…”

Section: In Vitro Conjugationmentioning

confidence: 99%

“…In a previous study, co-infection of mice with multiple β -lactam-resistant bacterial strains favoured colonization of Escherichia coli O80:H26 and enabled conjugative transfer of its bla CMY-2 gene via an IncI2 plasmid under ampicillin treatment [ 21 ]. Building on this model, we used the E. coli O80:H26 strain as a donor, along with a β -lactam-susceptible Salmonella Heidelberg, as a recipient in the present study.…”

Section: Introductionmentioning

confidence: 99%

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Mobility of β-lactam resistance under ampicillin treatment in gut microbiota suffering from pre-disturbance

et al. 2021

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Ingestion of food- or waterborne antibiotic-resistant bacteria may lead to dissemination of antibiotic resistance genes (ARGs) in the gut microbiota. The gut microbiota often suffers from various disturbances. It is not clear whether and how disturbed microbiota may affect ARG mobility under antibiotic treatments. For proof of concept, in the presence or absence of streptomycin pre-treatment, mice were inoculated orally with a β-lactam-susceptible Salmonella enterica serovar Heidelberg clinical isolate (recipient) and a β-lactam resistant Escherichia coli O80:H26 isolate (donor) carrying a blaCMY-2 gene on an IncI2 plasmid. Immediately following inoculation, mice were treated with or without ampicillin in drinking water for 7 days. Faeces were sampled, donor, recipient and transconjugant were enumerated, blaCMY-2 abundance was determined by quantitative PCR, faecal microbial community composition was determined by 16S rRNA amplicon sequencing and cecal samples were observed histologically for evidence of inflammation. In faeces of mice that received streptomycin pre-treatment, the donor abundance remained high, and the abundance of S. Heidelberg transconjugant and the relative abundance of Enterobacteriaceae increased significantly during the ampicillin treatment. Co-blooming of the donor, transconjugant and commensal Enterobacteriaceae in the inflamed intestine promoted significantly (P<0.05) higher and possibly wider dissemination of the blaCMY-2 gene in the gut microbiota of mice that received the combination of streptomycin pre-treatment and ampicillin treatment (Str–Amp) compared to the other mice. Following cessation of the ampicillin treatment, faecal shedding of S. Heidelberg transconjugant persisted much longer from mice in the Str–Amp group compared to the other mice. In addition, only mice in the Str–Amp group shed a commensal E. coli O2:H6 transconjugant, which carries three copies of the blaCMY-2 gene, one on the IncI2 plasmid and two on the chromosome. The findings highlight the significance of pre-existing gut microbiota for ARG dissemination and persistence during and following antibiotic treatments of infectious diseases.

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Section: Methodsmentioning

confidence: 98%

Section: Methodsmentioning

confidence: 99%

Section: Whole Genome Sequencingmentioning

confidence: 99%

Section: In Vitro Conjugationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Mobility of β-lactam resistance under ampicillin treatment in gut microbiota suffering from pre-disturbance

et al. 2021

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Transfer dynamics of antimicrobial resistance among gram-negative bacteria

Wang,

Farhan,

Yuan

et al. 2024

Science of The Total Environment

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An optogenetic toolkit for light-inducible antibiotic resistance

2023

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Antibiotics are a key control mechanism for synthetic biology and microbiology. Resistance genes are used to select desired cells and regulate bacterial populations, however their use to-date has been largely static. Precise spatiotemporal control of antibiotic resistance could enable a wide variety of applications that require dynamic control of susceptibility and survival. Here, we use light-inducible Cre recombinase to activate expression of drug resistance genes in Escherichia coli. We demonstrate light-activated resistance to four antibiotics: carbenicillin, kanamycin, chloramphenicol, and tetracycline. Cells exposed to blue light survive in the presence of lethal antibiotic concentrations, while those kept in the dark do not. To optimize resistance induction, we vary promoter, ribosome binding site, and enzyme variant strength using chromosome and plasmid-based constructs. We then link inducible resistance to expression of a heterologous fatty acid enzyme to increase production of octanoic acid. These optogenetic resistance tools pave the way for spatiotemporal control of cell survival.

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Mobility of β-Lactam Resistance Under Bacterial Co-infection and Ampicillin Treatment in a Mouse Model

Cited by 8 publications

References 50 publications

Mobility of β-lactam resistance under ampicillin treatment in gut microbiota suffering from pre-disturbance

Mobility of β-lactam resistance under ampicillin treatment in gut microbiota suffering from pre-disturbance

Transfer dynamics of antimicrobial resistance among gram-negative bacteria

An optogenetic toolkit for light-inducible antibiotic resistance

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