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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 Cover

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

Journal of Veterinary Research
Volume 67 (2023): Issue 4 (December 2023)
By: ,  ,   and    
Open Access
|Dec 2023
Figures & tables

Figures & Tables

Table 1.

Characteristics of the tested microorganisms

BacteriaBacterial features (reference)PhageFamilyPhage features
Pseudomonas syringae DSM 21482Isolated from diseased plants, one of the best-studied plant pathogens (43)Phi6 (Φ6)CystoviridaeSize ~ 80–100 nm; dsRNA; enveloped (lipid membrane), medium sized, no tail
Escherichia coli DSM 5695Pathogenic strains can cause distinct disease syndromes, are common food contaminants (23)MS2LeviviridaeSize ~ 23–28 nm; ssRNA; non-enveloped, small size and genome, no tail
Escherichia coli DSM 613T4MyoviridaeSize ~ 120–200 nm / 86–90 nm; dsDNA; nonenveloped, relatively big, contractile tail

1 DSM – bacterium from the Leibniz Institute Deutsche Sammlung von Mikroorganismen und Zellkulturen

Table 2.

Chosen treatment combinations for real time assessment of bacterial growth

Bacterial hostPhagePhage titreExtractExtract (%)Treatment type
P. syringae DSM 21482Phi6108SRa25Phage + extract
P. syringae DSM 21482--SRa25Extract
P. syringae DSM 21482Phi6108SRa0.049P + E + B combination *
P. syringae DSM 21482Phi6108--Phage infection control
P. syringae DSM 21482----Growth control
E. coli DSM 5695MS2108SRa25Phage + extract
E. coli DSM 5695--SRa25Extract
E. coli DSM 5695MS2108SRa0.19P + E + B combination *
E. coli DSM 5695MS2108--Phage infection control
E. coli DSM 5695----Growth control
E. coli DSM 613T4108SRa25Phage + extract
E. coli DSM 613--SRa25Extract
E. coli DSM 613T4104SRa0.049P + E + B combination *
E. coli DSM 613T4108--Phage infection control
E. coli DSM 613----Growth control
P. syringae DSM 21482Phi6108SRm25Phage + extract
P. syringae DSM 21482--SRm25Extract
P. syringae DSM 21482Phi6108SRm0.049P + E + B combination *
P. syringae DSM 21482Phi6108--Phage infection control
P. syringae DSM 21482----Growth control
E. coli DSM 5695MS2108SRm25Phage + extract
E. coli DSM 5695--SRm25Extract
E. coli DSM 5695MS2108SRm0.19P + E + B combination *
E. coli DSM 5695MS2108--Phage infection control
E. coli DSM 5695----Growth control
E. coli DSM 613T4108SRm25Phage + extract
E. coli DSM 613--SRm25Extract
E. coli DSM 613T4108SRm0.19P + E + B combination *
E. coli DSM 613T4108--Phage infection control
E. coli DSM 613----Growth control

1 DSM – bacterium from the Leibniz Institute Deutsche Sammlung von Mikroorganismen und Zellkulturen; SRa – S. rebaudiana acetone extract; SRm – S. rebaudiana methanol extract

1* P + E + B combination – phage, extract and bacteria combination in which interesting phenomena were detected

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

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

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

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

DOI: https://doi.org/10.2478/jvetres-2023-0059 | Journal eISSN: 2450-8608 (formerly 2300-3235)
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Language: English
Page range: 545 - 557
Submitted on: May 13, 2023
Accepted on: Oct 12, 2023
Published on: Dec 19, 2023
Published by: National Veterinary Research Institute in Pulawy
In partnership with: Paradigm Publishing Services
Keywords:
bacteriophage,
Stevia rebaudiana,
plant extract,
co-application
Related subjects:
Life sciences,
Molecular biology,
Microbiology and virology,
Life sciences, other,
Medicine,
Veterinary medicine

© 2023 Xymena Stachurska, Małgorzata Mizielińska, Magdalena Ordon, Paweł Nawrotek, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Volume 67 (2023): Issue 4 (December 2023)
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