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Isolation and characterization of biofilm-disrupting proteus phage Premi
Guadalupe Valencia-Toxqui1, Shobana Sugumar2, Jolene Ramsey3
1Department of Biology, Center for Phage Technology, Texas A&M University, College Station, TX, USA.
Abstract:
Proteus mirabilis is a biofilm-forming, multidrug-resistant bacterium and one of the most common causes of catheter-associated urinary tract infections (CAUTIs). Phage therapy is an alternative method that can be used to address the problem of multidrug-resistance. In this study, we report isolation and characterization of virulent phage Premi. The phage exhibits lytic activity against 4 out of 30 clinical isolates of P. mirabilis tested and is stable when exposed to pH values between 3 and 11. Phage Premi demonstrated significant anti-biofilm activity against P. mirabilis, reducing 24-hour established biofilms by 59-68%. In the 42-kb Premi genome, functions were assigned for the 50 predicted protein-coding genes including those involved in DNA replication, DNA modification, and lysis. Structural proteins were verified using mass spectrometry of purified virions. A comparison of its genomic features and phylogenetic analysis revealed that phage Premi is a podophage member of the order Caudoviricetes sharing 96% nucleotide similarity with Proteus phage PM 85 and has a T7-like phage genomic organization. Our study shows that Premi effectively inhibits P. mirabilis biofilms and could be a promising antimicrobial agent for treating drug-resistant P. mirabilis infection.
Insights
Phage Premi effectively combats Proteus mirabilis biofilms, a common cause of catheter-associated urinary tract infections. This virulent phage shows promise as an alternative antimicrobial for multidrug-resistant bacterial infections.
Area of Science:
- Microbiology
- Bacteriology
- Virology
Background:
- Proteus mirabilis is a multidrug-resistant bacterium causing catheter-associated urinary tract infections (CAUTIs).
- Biofilm formation by P. mirabilis complicates treatment and contributes to CAUTI persistence.
- Phage therapy offers a potential alternative to antibiotics for combating resistant bacteria.
Purpose of the Study:
- To isolate and characterize a virulent bacteriophage, named Premi, targeting P. mirabilis.
- To evaluate the anti-biofilm efficacy of phage Premi against P. mirabilis.
- To analyze the genomic features and phylogenetic relationship of phage Premi.
Main Methods:
- Isolation and characterization of virulent phage Premi from P. mirabilis.
- Assessment of phage lytic activity against clinical P. mirabilis isolates.
- Determination of phage stability across a range of pH values.
- Quantification of anti-biofilm activity using established biofilms.
- Whole-genome sequencing and bioinformatic analysis of phage Premi.
- Mass spectrometry analysis of purified phage virions.
Main Results:
- Phage Premi demonstrated lytic activity against 4 of 30 P. mirabilis clinical isolates.
- The phage remained stable at pH values ranging from 3 to 11.
- Phage Premi significantly reduced established P. mirabilis biofilms by 59-68%.
- The 42-kb genome of Premi contains 50 predicted protein-coding genes involved in replication, modification, and lysis.
- Phylogenetic analysis identified Premi as a podophage, closely related to Proteus phage PM 85, with a T7-like genomic organization.
Conclusions:
- Phage Premi exhibits potent anti-biofilm activity against P. mirabilis.
- Premi's stability and genomic characteristics suggest its potential as a therapeutic agent.
- This study highlights phage Premi as a promising candidate for combating drug-resistant P. mirabilis infections, particularly CAUTIs.
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