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The use of plant extracts and bacteriophages as an alternative therapy approach in combatting bacterial infections: the study of lytic phages and Stevia rebaudiana

Xymena Stachurska
Xymena Stachurska
, 
Małgorzata Mizielińska
Małgorzata Mizielińska
, 
Magdalena Ordon
Magdalena Ordon
 oraz 
Paweł Nawrotek
Paweł Nawrotek
   | 19 gru 2023
Journal of Veterinary Research's Cover Image
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Data publikacji: 19 gru 2023
Zakres stron: 545 - 557
Otrzymano: 13 maj 2023
Przyjęty: 12 paź 2023
DOI: https://doi.org/10.2478/jvetres-2023-0059
Słowa kluczowe
© 2023 Xymena Stachurska et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1.

Activity of the Stevia rebaudiana extracts on cells of E. coli and Pseudomonas syringae expressed in optical density (OD) changes of biomass. Acetone extract activity assessed by microdilution method (A) and methanol extract activity assessed by microdilution method (B). DSM – bacterium from the Leibniz Institute Deutsche Sammlung von Mikroorganismen und Zellkulturen. Error bars represent standard deviation between samples. Means sharing the asterisk are significantly different from the control (extract concentration 0%) at P ≤ 0.05
Activity of the Stevia rebaudiana extracts on cells of E. coli and Pseudomonas syringae expressed in optical density (OD) changes of biomass. Acetone extract activity assessed by microdilution method (A) and methanol extract activity assessed by microdilution method (B). DSM – bacterium from the Leibniz Institute Deutsche Sammlung von Mikroorganismen und Zellkulturen. Error bars represent standard deviation between samples. Means sharing the asterisk are significantly different from the control (extract concentration 0%) at P ≤ 0.05

Fig. 2.

Phage titres (MS2, Phi6 and T4) after 24 h exposure to different concentrations of S. rebaudiana extracts. Phage counts after co-incubation of phages with acetone extract (A) and after co-incubation of phages with methanol extract (B). Error bars represent standard deviation between samples. Means sharing the asterisk are significantly different from the control at P ≤ 0.05. PFU – plaque-forming unit
Phage titres (MS2, Phi6 and T4) after 24 h exposure to different concentrations of S. rebaudiana extracts. Phage counts after co-incubation of phages with acetone extract (A) and after co-incubation of phages with methanol extract (B). Error bars represent standard deviation between samples. Means sharing the asterisk are significantly different from the control at P ≤ 0.05. PFU – plaque-forming unit

Fig. 3.

Effects of phage–extract combinations on host bacteria. Paired heat-maps represent optical density measurements at 600 nm (A–F) and corresponding fluorescence measurements (A’–F’). Combined treatment of Stevia rebaudiana acetone extract with phage Phi6 (A–A’), phage MS2 (B–B’) and phage T4 (C–C’). Combined treatment of S. rebaudiana methanol extract with phage Phi6 (D–D’), phage MS2 (E–E’) and phage T4 (F–F’). Synograms (t = 24 h) represent the mean reduction (% of the control) or activity (% of the control) percentage of each treatment from three replicates. DSM – bacterium from the Leibniz Institute Deutsche Sammlung von Mikroorganismen und Zellkulturen; PFU – plaque-forming unit
Effects of phage–extract combinations on host bacteria. Paired heat-maps represent optical density measurements at 600 nm (A–F) and corresponding fluorescence measurements (A’–F’). Combined treatment of Stevia rebaudiana acetone extract with phage Phi6 (A–A’), phage MS2 (B–B’) and phage T4 (C–C’). Combined treatment of S. rebaudiana methanol extract with phage Phi6 (D–D’), phage MS2 (E–E’) and phage T4 (F–F’). Synograms (t = 24 h) represent the mean reduction (% of the control) or activity (% of the control) percentage of each treatment from three replicates. DSM – bacterium from the Leibniz Institute Deutsche Sammlung von Mikroorganismen und Zellkulturen; PFU – plaque-forming unit

Fig. 4.

Bacteria growth curves treated by lytic Phi6, MS2 and T4 phages and different extract concentrations chosen in the synograms assay. Combined treatment of phages with of Stevia rebaudiana acetone extract (Phi6 – A; MS2 – A’; T4 – A”) and combined treatment of phages with Stevia rebaudiana methanol extract (Phi6 – B; MS2 – B’; T4 – B’’). OD – optical density; DSM – bacterium from the Leibniz Institute Deutsche Sammlung von Mikroorganismen und Zellkulturen
Bacteria growth curves treated by lytic Phi6, MS2 and T4 phages and different extract concentrations chosen in the synograms assay. Combined treatment of phages with of Stevia rebaudiana acetone extract (Phi6 – A; MS2 – A’; T4 – A”) and combined treatment of phages with Stevia rebaudiana methanol extract (Phi6 – B; MS2 – B’; T4 – B’’). OD – optical density; DSM – bacterium from the Leibniz Institute Deutsche Sammlung von Mikroorganismen und Zellkulturen

Characteristics of the tested microorganisms

Bacteria Bacterial features (reference) Phage Family Phage features
Pseudomonas syringae DSM 21482 Isolated from diseased plants, one of the best-studied plant pathogens (43) Phi6 (Φ6) Cystoviridae Size ~ 80–100 nm; dsRNA; enveloped (lipid membrane), medium sized, no tail
Escherichia coli DSM 5695 Pathogenic strains can cause distinct disease syndromes, are common food contaminants (23) MS2 Leviviridae Size ~ 23–28 nm; ssRNA; non-enveloped, small size and genome, no tail
Escherichia coli DSM 613 T4 Myoviridae Size ~ 120–200 nm / 86–90 nm; dsDNA; nonenveloped, relatively big, contractile tail

Chosen treatment combinations for real time assessment of bacterial growth

Bacterial host Phage Phage titre Extract Extract (%) Treatment type
P. syringae DSM 21482 Phi6 108 SRa 25 Phage + extract
P. syringae DSM 21482 - - SRa 25 Extract
P. syringae DSM 21482 Phi6 108 SRa 0.049 P + E + B combination *
P. syringae DSM 21482 Phi6 108 - - Phage infection control
P. syringae DSM 21482 - - - - Growth control
E. coli DSM 5695 MS2 108 SRa 25 Phage + extract
E. coli DSM 5695 - - SRa 25 Extract
E. coli DSM 5695 MS2 108 SRa 0.19 P + E + B combination *
E. coli DSM 5695 MS2 108 - - Phage infection control
E. coli DSM 5695 - - - - Growth control
E. coli DSM 613 T4 108 SRa 25 Phage + extract
E. coli DSM 613 - - SRa 25 Extract
E. coli DSM 613 T4 104 SRa 0.049 P + E + B combination *
E. coli DSM 613 T4 108 - - Phage infection control
E. coli DSM 613 - - - - Growth control
P. syringae DSM 21482 Phi6 108 SRm 25 Phage + extract
P. syringae DSM 21482 - - SRm 25 Extract
P. syringae DSM 21482 Phi6 108 SRm 0.049 P + E + B combination *
P. syringae DSM 21482 Phi6 108 - - Phage infection control
P. syringae DSM 21482 - - - - Growth control
E. coli DSM 5695 MS2 108 SRm 25 Phage + extract
E. coli DSM 5695 - - SRm 25 Extract
E. coli DSM 5695 MS2 108 SRm 0.19 P + E + B combination *
E. coli DSM 5695 MS2 108 - - Phage infection control
E. coli DSM 5695 - - - - Growth control
E. coli DSM 613 T4 108 SRm 25 Phage + extract
E. coli DSM 613 - - SRm 25 Extract
E. coli DSM 613 T4 108 SRm 0.19 P + E + B combination *
E. coli DSM 613 T4 108 - - Phage infection control
E. coli DSM 613 - - - - Growth control
eISSN:
2450-8608
Język:
Angielski
Częstotliwość wydawania:
4 razy w roku
Dziedziny czasopisma:
Life Sciences, Molecular Biology, Microbiology and Virology, other, Medicine, Veterinary Medicine
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