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Comparative Methodological Evaluation of Three Bacteriophage Isolation Methods From Environmental Water Samples
Mohammod Johirul Islam1, Mohammad Kamruzzaman1, Fahim Alam Nobel1
1Department of Biochemistry and Molecular Biology, Mawlana Bhashani Science and Technology University, Tangail, BGD.
Cureus
|June 4, 2026
Summary
The host-based enrichment method significantly enhances bacteriophage (virus that infects bacteria) recovery from water samples compared to direct or PEG precipitation methods. This efficient technique is crucial for isolating phages for therapeutic and diagnostic applications.
Area of Science:
- Microbiology
- Virology
- Environmental Science
Background:
- Bacteriophages are viruses that infect bacteria, with growing importance in biocontrol, phage therapy, food safety, and diagnostics.
- Efficient isolation of bacteriophages from environmental sources is critical for research and applied uses.
- This study evaluates three methods for isolating bacteriophages from environmental water samples.
Purpose of the Study:
- To comparatively evaluate the efficiency of three bacteriophage isolation methods.
- To determine the most effective method for recovering bacteriophages from environmental water samples.
- To identify a robust strategy for isolating bacteriophages for various applications.
Main Methods:
- Collected 96 environmental water samples over one year from four aquatic sites in Bangladesh.
- Assessed six bacteriophages targeting clinically relevant bacteria using direct recovery, polyethylene glycol (PEG)-mediated precipitation, and host-based enrichment.
- Compared phage recovery efficiency using statistical tests and estimated effect sizes with odds ratios.
Main Results:
- The host-based enrichment method achieved the highest average phage recovery rate (47.20%), significantly outperforming PEG precipitation (21.35%) and direct recovery (9.38%).
- Statistical analysis confirmed a significant difference in recovery proportions among the three methods (p < 0.0001).
- Pairwise comparisons showed enrichment yielded significantly higher recovery than PEG (OR=3.29) and direct methods (OR=8.65).
Conclusions:
- The modified host-based enrichment method significantly improves bacteriophage recovery from environmental water samples.
- This method is particularly effective for isolating low-abundance phages, offering a robust and sensitive strategy.
- The findings support the use of enrichment for advancing phage-based therapeutics, environmental surveillance, and microbial control.
