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. 2024 Feb 19;13(2):195.
doi: 10.3390/antibiotics13020195.

Potential of an Isolated Bacteriophage to Inactivate Klebsiella pneumoniae: Preliminary Studies to Control Urinary Tract Infections

João Duarte  1 , Carolina Máximo  1 , Pedro Costa  1 , Vanessa Oliveira  1 , Newton C M Gomes  1 , Jesús L Romalde  2 , Carla Pereira  1 , Adelaide Almeida  1
Affiliations

Potential of an Isolated Bacteriophage to Inactivate Klebsiella pneumoniae: Preliminary Studies to Control Urinary Tract Infections

João Duarte et al. Antibiotics (Basel). .

Abstract

Urinary tract infections (UTIs) caused by resistant Klebsiella pneumoniae can lead to severe clinical complications and even death. An alternative treatment option for infected patients is using bacteriophages. In the present study, we isolated phage VB_KPM_KP1LMA (KP1LMA) from sewage water using a K. pneumoniae strain as a host. Whole-genome analysis indicated that the genome was a double-stranded linear 176,096-bp long DNA molecule with 41.8% GC content and did not contain virulence or antibiotic resistance genes. The inactivation potential of phage KP1LMA was assessed in broth at an MOI of 1 and 10, and a maximum inactivation of 4.9 and 5.4 log CFU/mL, respectively, was observed after 9 h. The efficacy at an MOI of 10 was also assessed in urine to evaluate the phage's performance in an acidic environment. A maximum inactivation of 3.8 log CFU/mL was observed after 9 h. The results suggest that phage KP1LMA could potentially control a UTI caused by this strain of K. pneumoniae, indicating that the same procedure can be used to control UTIs caused by other strains if new specific phages are isolated. Although phage KP1LMA has a narrow host range, in the future, efforts can be made to expand its spectrum of activity and also to combine this phage with others, potentially enabling its use against other K. pneumoniae strains involved in UTIs.

Keywords: Klebsiella pneumoniae; bacteriophage; phage therapy; urinary tract infections.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Phage plaque morphology (A) and electron micrograph photography (B).
Figure 2
Figure 2
The genome map of phage KP1LMA. The outer circle with arrow-headed bands represents the coding DNA sequences (CDS) color coded according to the functional category of the predicted gene in the direction of the transcription. The innermost ring represents the genome GC skew (green/pink) followed by GC content (black/grey). The labels show the predicted functions of the functional CDSs, color-coded by the PHROGs category (Supplementary Table S1).
Figure 3
Figure 3
Adsorption curves of phages KP1LMA in the presence of K. pneumoniae as the host. Values represent the mean of three independent experiments, and error bars represent the standard deviation.
Figure 4
Figure 4
One-step growth curve of phage KP1LMA in the presence of the host, K. pneumoniae. Values represent the mean of three independent experiments, and error bars represent the standard deviation.
Figure 5
Figure 5
Inactivation of K. pneumoniae using phage KP1LMA at two different MOIs. (A) Bacterial concentrations: BC—bacterial control; BP MOI 1—bacteria plus phage at an MOI of 1; BP MOI 10—bacteria plus phage at an MOI of 10. (B) Phage concentration: PC MOI 1—phage control at an MOI of 1; PC MOI 10: phage control at an MOI of 10; PB MOI 1—phage and bacteria at an MOI of 1; PB MOI 10—phage and bacteria at an MOI of 10. The experiments were performed in TSB at 37 °C, and values represent the mean of three independent experiments. Error bars represent the standard deviation.
Figure 6
Figure 6
Inactivation of K. pneumoniae in sterile urine by phage KP1LMA using an MOI of 10. (A): bacterial concentrations; bacteria control—bacteria in the absence of phage; BP MOI 10—bacteria plus phage at an MOI of 10. (B): Phage concentration; PC MOI 10—phage control; PB MOI 10—phage in the presence of bacteria at a final MOI of 10. Values represent the mean of three independent experiments. Error bars represent the standard deviation.
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Through national funds, we acknowledge financial support to CESAM from FCT/MCTES (UIDP/50017/2020+UIDB/50017/2020+LA/P/0094/2020).

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