Disrupting the mixed-species biofilm of Klebsiella pneumoniae B5055 and Pseudomonas aeruginosa PAO using bacteriophages alone or in combination with xylitol

Abstract

We investigated the potential of bacteriophages alone as well as in combination with xylitol for tackling mixed-species biofilm of Pseudomonas aeruginosa and Klebsiella pneumoniae. When mixed-species biofilm was established on polycarbonate discs, P. aeruginosa formed the base layer which was physically shielded on the top by K. pneumoniae. Thereafter, mixed-species biofilm was treated with bacteriophages. K. pneumoniae-specific depolymerase-producing phage KPO1K2 caused significant reduction in the count of Klebsiella. In contrast, P. aeruginosaspecific non-depolymerase-producing phage Pa29 failed to cause any reduction in the count of Pseudomonas. However, application of both phages together resulted in significant reduction in the count of both organisms. This suggests that depolymerase produced by phage KPO1K2 hydrolysed the top layer of K. pneumoniae and guided the entry of Pa29 to reach P. aeruginosa lying underneath. This phenomenon was confirmed when K. pneumoniae-specific nondepolymerase-producing phage NDP was used along with Pa29. Pa29 could not penetrate and reach its host bacterium. Xylitol worked synergistically along with the phage, resulting in a significant decrease in counts of both organisms. Disruption of mixed species biofilm by phage and xylitol was confirmed on the basis of the amount of protein and DNA released. This phagebased approach to altering the structural pattern and disrupting the mixed species biofilm is the first of its kind. It can be used as a topical application, a coating for foreign bodies or for aerosol delivery to tackle infections where both pathogens coexist in a biofilm mode.

Using Bacteriophages as a Trojan Horse to the Killing of Dual-Species Biofilm Formed by Pseudomonas aeruginosa and Methicillin Resistant Staphylococcus aureus

“…In this scenario, bacteriophages appear to be an alternative strategy to treat biofilm-forming infections. Over the past few years, numerous studies have been carried out investigating the effectiveness of phages against mono-and dualspecies biofilm (Sillankorva et al, 2010;Chhibber et al, 2015;Gutierrez et al, 2015;Gonzalez et al, 2017;Melo et al, 2019). Many of these studies pointed to a notable dependency between the phages and the bacterial species involved on the biofilm for the efficacy of the phage treatment.…”

“…Bacteriophages (phages) have regained interest as promising therapeutic option in fighting biofilmrelated infections due to their rapid bactericidal activity, the self-amplification ability and potential biofilm degradative properties (Harper et al, 2014). However, there are only limited studies investigating the activity of phages against polymicrobial biofilms (Sillankorva et al, 2010;Kay et al, 2011;Chhibber et al, 2015;Oliveira et al, 2018;Melo et al, 2019) and just recently Akturk et al (2019) evaluated the simultaneous and staggered administration of a P. aeruginosa-targeting monophage and conventional antibiotics on S. aureus/P. aeruginosa dualspecies biofilms.…”

A Novel Approach for Combating Klebsiella pneumoniae Biofilm Using Histidine Functionalized Silver Nanoparticles

“…Biofilm age is a major decisive factor in determining the effectiveness of any antibiofilm agent (Sedlacek and Walker, 2007; Ito et al, 2009; Chhibber et al, 2015). As biofilm grows older it becomes difficult to eradicate them by conventional and newer antibiotics due to their altered metabolic activity and thicker extracellular matrix (Rani et al, 2007).…”

Immunological Tolerance and Function: Associations Between Intestinal Bacteria, Probiotics, Prebiotics, and Phages

“…Bacteriophage directed control of behaviors of how bacteria colonize and survive in different anatomical sites encourage developmental properties that helps establish commensal populations that reduces the risk of disease ( 182 ). Studies report that bacteriophages may be involved in important functions in human immunity by defending the intestinal epithelial barrier and mucosal tissue from infections by pathobionts ( 188 ).…”