Campylobacter jejuni 11168H Exposed to Penicillin Forms Persister Cells and Cells With Altered Redox Protein Activity

Morcrette, Helen; Kovacs-Simon, Andrea; Tennant, Richard K.; Love, John; Wagley, Sariqa; Yang, Zheng; Studholme, David J.; Soyer, Orkun S.; Champion, Olivia L.; Butler, Catherine; Titball, Richard W.

doi:10.3389/fcimb.2020.565975

Cited by 12 publications

(9 citation statements)

References 55 publications

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“…However, there was no size distinction between metabolically active and inactive cells in L. plantarum ( Figures 2G, H ). Interestingly, peaks in mean RSG intensities in stationary phase (as well as exponential phase) may highlight a shift in metabolic activity ( Figures 2B , 3B ), corresponding to a stress response ( Morcrette et al., 2020 ).…”

Section: Discussionmentioning

confidence: 99%

The Application of Imaging Flow Cytometry for Characterisation and Quantification of Bacterial Phenotypes

Power

Barber

Groenhof

et al. 2021

Front. Cell. Infect. Microbiol.

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Bacteria modify their morphology in response to various factors including growth stage, nutrient availability, predation, motility and long-term survival strategies. Morphological changes may also be associated with specific physiological phenotypes such as the formation of dormant or persister cells in a “viable but non-culturable” (VBNC) state which frequently display different shapes and size compared to their active counterparts. Such dormancy phenotypes can display various degrees of tolerance to antibiotics and therefore a detailed understanding of these phenotypes is crucial for combatting chronic infections and associated diseases. Cell shape and size are therefore more than simple phenotypic characteristics; they are important physiological properties for understanding bacterial life-strategies and pathologies. However, quantitative studies on the changes to cell morphologies during bacterial growth, persister cell formation and the VBNC state are few and severely constrained by current limitations in the most used investigative techniques of flow cytometry (FC) and light or electron microscopy. In this study, we applied high-throughput Imaging Flow Cytometry (IFC) to characterise and quantify, at single-cell level and over time, the phenotypic heterogeneity and morphological changes in cultured populations of four bacterial species, Bacillus subtilis, Lactiplantibacillus plantarum, Pediococcus acidilactici and Escherichia coli. Morphologies in relation to growth stage and stress responses, cell integrity and metabolic activity were analysed. Additionally, we were able to identify and morphologically classify dormant cell phenotypes such as VBNC cells and monitor the resuscitation of persister cells in Escherichia coli following antibiotic treatment. We therefore demonstrate that IFC, with its high-throughput data collection and image capture capabilities, provides a platform by which a detailed understanding of changes in bacterial phenotypes and their physiological implications may be accurately monitored and quantified, leading to a better understanding of the role of phenotypic heterogeneity in the dynamic microbiome.

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Section: Discussionmentioning

confidence: 99%

The Application of Imaging Flow Cytometry for Characterisation and Quantification of Bacterial Phenotypes

Power

Barber

Groenhof

et al. 2021

Front. Cell. Infect. Microbiol.

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“…Streptomycin treatment shifted the majority of the intact Pph population to a state of increased RSG staining intensity. Antibiotics stimulate the production of reactive oxygen species (ROS) in bacteria ( 46 ), and antibiotic-induced increases in RSG intensity were recently associated with an accumulation of ROS-protective reductases in Campylobacter jejuni ( 47 ). Thus, we suspect that the high-redox fraction in Pph cells similarly reflects a reductase response to intracellular ROS production, although ROS-specific methods would be needed to determine this conclusively.…”

Section: Discussionmentioning

confidence: 99%

Pseudomonas syringae pv. phaseolicola Uses Distinct Modes of Stationary-Phase Persistence To Survive Bacteriocin and Streptomycin Treatments

Patel

Kandel

Traverso

et al. 2021

mBio

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Antimicrobial treatment of bacteria often results in a small population of surviving tolerant cells, or persisters, that may contribute to recurrent infection. Antibiotic persisters are metabolically dormant, but the basis of their persistence in the presence of membrane-disrupting biological compounds is less well understood. We previously found that the model plant pathogen Pseudomonas syringae pv. phaseolicola 1448A (Pph) exhibits persistence to tailocin, a membrane-disrupting biocontrol compound with potential for sustainable disease control. Here, we compared physiological traits associated with persistence to tailocin and to the antibiotic streptomycin and established that both treatments leave similar frequencies of persisters. Microscopic profiling of treated populations revealed that while tailocin rapidly permeabilizes most cells, streptomycin treatment results in a heterogeneous population in the redox and membrane permeability state. Intact cells were sorted into three fractions according to metabolic activity, as indicated by a redox-sensing reporter dye. Streptomycin persisters were cultured from the fraction associated with the lowest metabolic activity, but tailocin persisters were cultured from a fraction associated with an active metabolic signal. Cells from culturable fractions were able to infect host plants, while the nonculturable fractions were not. Tailocin and streptomycin were effective in eliminating all persisters when applied sequentially, in addition to eliminating cells in other viable states. This study identifies distinct metabolic states associated with antibiotic persistence, tailocin persistence, and loss of virulence and demonstrates that tailocin is highly effective in eliminating dormant cells. IMPORTANCE Populations of genetically identical bacteria encompass heterogeneous physiological states. The small fraction of bacteria that are dormant can help the population survive exposure to antibiotics and other stresses, potentially contributing to recurring infection cycles in animal or plant hosts. Membrane-disrupting biological control treatments are effective in killing dormant bacteria, but these treatments also leave persister-like survivors. The current work demonstrates that in Pph, persisters surviving treatment with membrane-disrupting tailocin proteins have an elevated redox state compared to that of dormant streptomycin persisters. Combination treatment was effective in killing both persister types. Culturable persisters corresponded closely with infectious cells in each treated population, whereas the high-redox and unculturable fractions were not infectious. In linking redox states to heterogeneous phenotypes of tailocin persistence, streptomycin persistence, and infection capability, this work will inform the search for mechanisms and markers for each phenotype.

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“…The authors attribute this increase to the presence of persistent, metabolically dormant cells, usually present in biofilms. These cells, recently preceded in C. jejuni , are extremely tolerant to antibiotics without undergoing any genetic alteration ( Morcrette et al., 2020 ). Another reason for this was suggested in a previous study, which proposed that some antibiotics can act as antagonists of the formation (and growth) of biofilm in low concentrations, agonists in higher concentrations and antagonists in even higher levels ( Kaplan, 2011 ).…”

Section: Discussionmentioning

confidence: 99%

Antibiotic Resistance in the Alternative Lifestyles of Campylobacter jejuni

Rossi

Dumont

Santos

et al. 2021

Front. Cell. Infect. Microbiol.

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Campylobacter jejuni is the main pathogen identified in cases of foodborne gastroenteritis worldwide. Its importance in poultry production and public health is highlighted due to the growing antimicrobial resistance. Our study comparatively investigated the effect of five different classes of antimicrobials on the planktonic and biofilm forms of 35 strains of C. jejuni with high phylogenetic distinction in 30 of them. In the planktonic form, the existence of susceptible strains to colistin (7/35 – 20%) and resistance to meropenem (3/35 – 8.6%) represent a novelty in strains evaluated in Brazil. In biofilms formed with the addition of chicken juice, the number of resistant strains was significantly higher for colistin, erythromycin and meropenem (100%), but the susceptibility to tetracycline was shown as a control strategy for specific cases. High concentrations (1,060 ± 172.1mg/L) of antibiotics were necessary to control the biofilm structure in susceptible strains in the planktonic form, which is consistent with the high biomass produced in these strains. Stainless steel and polyurethane were the most (BFI=2.1) and least (BFI=1.6) favorable surfaces for the production of biomass treated with antimicrobials. It is concluded that the antimicrobial action was detected for all tested drugs in planktonic form. In sessile forms, the biomass production was intensified, except for tetracycline, which showed an antibiofilm effect.

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Campylobacter jejuni 11168H Exposed to Penicillin Forms Persister Cells and Cells With Altered Redox Protein Activity

Cited by 12 publications

References 55 publications

The Application of Imaging Flow Cytometry for Characterisation and Quantification of Bacterial Phenotypes

The Application of Imaging Flow Cytometry for Characterisation and Quantification of Bacterial Phenotypes

Pseudomonas syringae pv. phaseolicola Uses Distinct Modes of Stationary-Phase Persistence To Survive Bacteriocin and Streptomycin Treatments

Antibiotic Resistance in the Alternative Lifestyles of Campylobacter jejuni

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