Abstract
Recent studies have demonstrated that phages can be co-transported with motile non-host bacteria, thereby enabling their invasion of biofilms and control of biofilm composition. Here, we developed a novel approach to isolate non-host bacteria able to co-transport phages from soil. It is based on the capability of phage-carrying non-host bacteria to move along mycelia out of soil and form colonies in plaques of their co-transported phages. The approach was tested using two model phages of differing surface hydrophobicity, i.e., hydrophobic Escherichia virus T4 (T4) and hydrophilic Pseudoalteromonas phage HS2 (HS2). The phages were mixed into soil and allowed to be transported by soil bacteria along the mycelia of Pythium ultimum. Five phage-carrying bacterial species were isolated (Viridibacillus sp., Enterobacter sp., Serratia sp., Bacillus sp., Janthinobacterium sp.). These bacteria exhibited phage adsorption efficiencies of ≈90–95% for hydrophobic T4 and 30–95% for hydrophilic HS2. The phage adsorption efficiency of Viridibacillus sp. was ≈95% for both phages and twofold higher than T4-or HS2-adsorption to their respective hosts, qualifying Viridibacillus sp. as a potential super carrier for phages. Our approach offers an effective and target-specific way to identify and isolate phage-carrying bacteria in natural and man-made environments.
Highlights
Bacteriophages have received increasing interest as drivers of microbial ecology [1] and/or as agents to control bacterial biofilms relevant for human health [2,3], food hygiene [4,5], and environmental safety [6,7]
We evaluated the transport of bacteria and/or phages along mycelia in four scenarios: (i) PBS, (ii) T4 or HS2, (iii) P. putida KT2440, and (iv) T4 + P. putida KT2440 or HS2 + P. putida KT2440
We hypothesized that non-host bacteria can “pick up” phages and carry them along while dispersing along mycelia; this can be used for the targeted isolation of motile bacteria able to efficiently co-transport phages
Summary
Bacteriophages ( termed phages) have received increasing interest as drivers of microbial ecology [1] and/or as agents to control bacterial biofilms relevant for human health [2,3], food hygiene [4,5], and environmental safety [6,7]. Phages are intrinsically non-motile anti-bacterial agents that typically disperse by diffusion (diffusion ≈ 4 × 10−10 m s−1 [8,9]) or advection to their hosts. Phages may adsorbed to (bio-) surfaces and disperse together with their vectors (e.g., non-host bacteria [10] or invertebrates [11]). At short distances, they may be ejected by explosive cell lysis [12], while phages immobilized e.g., on soil particles can act as set-and-wait predators for dispersing host bacteria in soil [13,14]. Phage adsorption to environmental bacteria has been well described by multiple methods (e.g., metagenomic analysis [20], AdsorpSeq [27], fluorescent labeling [21], and cryo-electron microscopy [28]), few attempts have been made to isolate culturable phage-carrying bacteria as potential tools for challenging invasion hypothesis in microbial ecology or as phageassociated biofilm control agents in biotechnology
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