High-Resolution Bacterial Cytological Profiling Reveals Intrapopulation Morphological Variations upon Antibiotic Exposure

Samernate, Thanadon; Htoo, Htut Htut; Sugie, Joseph; Chavasiri, Warinthorn; Pogliano, Joe; Chaikeeratisak, Vorrapon; Nonejuie, Poochit

doi:10.1128/aac.01307-22

Cited by 8 publications

(6 citation statements)

References 73 publications

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“…This contradictory finding might be due to the fact that the checkerboard assay monitored the growth of the bacteria via an OD 600 reading, which correlates with the density of the bacterial culture, regardless of the size of the individual cell. Since it is well known that β-lactam-treated cells are much longer than the untreated cells ( 29 , 51 , 52 ), the OD 600 reading could not be accurately converted to cell viability count. Thus, cell viability count evaluation in each condition is crucial in determining the phage-antibiotic interaction, thereby improving the overall accuracy of defining the type of phage-antibiotic interaction.…”

Section: Discussionmentioning

confidence: 99%

A novel coli myophage and antibiotics synergistically inhibit the growth of the uropathogenic E. coli strain CFT073 in stoichiometric niches

Khunti,

Chantakorn,

Tantibhadrasapa

et al. 2023

Microbiol Spectr

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Urinary tract infections are widespread bacterial infections affecting millions of people annually, with Escherichia coli being the most prevalent. Although phage therapy has recently gained interest as a promising alternative therapy for antibiotic-resistant bacteria, several studies have raised concerns regarding the evolution of phage resistance, making the therapy ineffective. In this study, we discover a novel coli myophage designated as Killian that targets E. coli strains, including the uropathogenic E. coli (UPEC) strain CFT073. It requires at least 20 minutes for 90% of its particles to adsorb to the host cells, undergoes subcellular activities for replication for 30 minutes, and eventually lyses the cells with a burst size of about 139 particles per cell. Additionally, Killian can withstand a wide variety of temperatures (4–50°C) and pHs ( 4 – 10 ). Genome analysis reveals that Killian’s genome consists of 169,905 base pairs with 35.5% GC content, encoding 276 open reading frames; of these, 209 are functionally annotated with no undesirable genes detected, highlighting its potential as an antibiotic alternative against UPEC. However, after an 8-hour phage treatment at high multiplicities of infection, bacterial density continuously increases, indicating an onset of bacterial growth revival. Thus, the combination study between the phage and three different antibiotics, including amikacin, ciprofloxacin, and piperacillin, was performed and showed that certain pairs of phage and antibiotics exhibited synergistic interactions in suppressing the bacterial growth revival. These findings suggest that Killian-antibiotic combinations are effective in inhibiting the growth of UPEC. IMPORTANCE Phage therapy has recently been in the spotlight as a viable alternative therapy for bacterial infections. However, several studies have raised concerns about the emergence of phage resistance that occurs during treatment, making the therapy not much effective. Here, we present the discovery of a novel E. coli myophage that, by itself, can effectively kill the uropathogenic E. coli , but the emergence of bacterial growth revival was detected during the treatment. Phage and antibiotics are then combined to improve the efficiency of the phage in suppressing the bacterial re-growth. This research would pave the way for the future development of phage-antibiotic cocktails for the sustainable use of phages for therapeutic purposes.

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Section: Discussionmentioning

confidence: 99%

A novel coli myophage and antibiotics synergistically inhibit the growth of the uropathogenic E. coli strain CFT073 in stoichiometric niches

Khunti,

Chantakorn,

Tantibhadrasapa

et al. 2023

Microbiol Spectr

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“…32 It is also possible that L9 exhibits simultaneous inhibitory activity against multiple cellular pathways, resulting in a combinatorial effect on bacterial morphology, as observed previously when bacteria were treated with two antibiotics. 34,35 The data presented here identify the azobenzene G4 ligand L9 as possessing bacteriostatic antibacterial activity against reference and clinical multi-drug resistant E. coli. Electron and laser confocal microscopy, the latter supported by BCP analysis, show L9 treatment to induce a phenotype differing from those conferred by exposure to comparator antibiotics or G4 ligands, identifying L9 as an anti-Gram-negative agent with a distinct mode of action.…”

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confidence: 83%

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confidence: 99%

Small molecule G-quadruplex ligands are antibacterial candidates for Gram-negative bacteria

Ramos‐Soriano

Takebayashi

Samphire

et al. 2022

Preprint

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With the rise in bacterial antimicrobial resistance (AMR) and decline in antibiotic discovery, global healthcare is at a point of jeopardy. There is great need for the development of novel antimicrobials that target bacteria that have become resistant to our existing antibiotics, in particular Gram-negative species such as Escherichia coli which is responsible for opportunistic infections of already compromised patients. Here we demonstrate that a novel pyridinium-functionalised azobenzene scaffold L20, identified as a candidate ligand able target G-quadruplex (G4) structures in bacterial genomes/transcriptomes, shows promising antibacterial activity (MIC values ≤ 4 µg/ml) against multi-drug resistant E. coli. Tandem Mass Tag (TMT) proteomics applied to cultures of the E. coli type strain ATCC 25922 treated with sub-lethal concentrations of L20, identified seven G4-containing sequences as potential targets for L20. FRET (fluorescence resonance energy transfer) stabilisation assays indicate L20 binds these selected sequences with variable and moderate affinity, in contrast to two comparator G4 ligands (stiff-stilbene L5 and pyridostatin (PDS)) that better stabilise G4 structures but exhibit a lower antimicrobial activity. However, proteomic experiments also reveal that, alongside its superior antibacterial activity, L20 treatment influences expression levels of more G4-associated proteins than either L5 or PDS, and upregulates multiple essential proteins involved in translation. These findings identify strategies discovering potential G4 ligands as approaches that can lead to antibacterial candidates active against priority targets such as multi-drug resistant E. coli, and that targeting G4 sequences, and ligands such as L20, warrant further exploration as potential novel therapeutics with G4-mediated modes of action.

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“… 77 The morphological feature set was selected based on the previous study. 81 , 82 Statistical calculation and machine learning are carried out using the scikit-learn library in python. 83 Briefly, cell profile data were transformed and normalized with Cube Root Transformation and Z-scored normalization, 84 respectively.…”

Section: Methodsmentioning

confidence: 99%

Nucleus-forming jumbophage PhiKZ therapeutically outcompetes non-nucleus-forming jumbophage Callisto

Naknaen,

Samernate,

Saeju

et al. 2024

iScience

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High-Resolution Bacterial Cytological Profiling Reveals Intrapopulation Morphological Variations upon Antibiotic Exposure

Cited by 8 publications

References 73 publications

A novel coli myophage and antibiotics synergistically inhibit the growth of the uropathogenic E. coli strain CFT073 in stoichiometric niches

A novel coli myophage and antibiotics synergistically inhibit the growth of the uropathogenic E. coli strain CFT073 in stoichiometric niches

Small molecule G-quadruplex ligands are antibacterial candidates for Gram-negative bacteria

Nucleus-forming jumbophage PhiKZ therapeutically outcompetes non-nucleus-forming jumbophage Callisto

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