Differentiation of polyvalent bacteriophages specific to uropathogenic Proteus mirabilis strains based on the host range pattern and RFLP.

Abstract

Urinary tract infections (UTIs) caused by P. mirabilis are difficult to cure because of the increasing antimicrobial resistance of these bacteria. Phage therapy is proposed as an alternative infection treatment. The aim of this study was to isolate and differentiate uropathogenic P. mirabilis strain specific polyvalent bacteriophages producing polysaccharide depolymerases (PDs). 51 specific phages were obtained. The plaques of 29 bacteriophages were surrounded by halos, which indicated that they produced PDs. The host range analysis showed that, except phages 58B and 58C, the phage host range profiles differed from each other. Phages 35 and 45 infected all P. mirabilis strains tested. Another 10 phages lysed more than 90% of isolates. Among these phages, 65A, 70, 66 and 66A caused a complete lysis of the bacterial lawn formed by 62% to 78% of strains. Additionally, phages 39A and 70 probably produced PDs. The phages' DNA restriction fragment length polymorphism (RFLP) analysis demonstrated that genomes of 51 isolated phages represented 34 different restriction profiles. DNA of phage 58A seemed to be resistant to selected EcoRV endonuclease. The 33 RFLP-EcoRV profiles showed a Dice similarity index of 38.8%. 22 RFLP patterns were obtained from single phage isolates. The remaining 12 restriction profiles consisted of 2 to 4 viruses. The results obtained from phage characterization based on the pattern of phage host range in combination with the RFLP method enabled effective differentiation of the studied phages and selection of PD producing polyvalent phages for further study.

Isolation and Characterization of a Lytic Bacteriophage (vB_PmiS-TH) and Its Application in Combination with Ampicillin against Planktonic and Biofilm Forms of Proteus mirabilis Isolated from Urinary Tract Infection

“…P. mirabilis accounts for 12% of complicated UTIs, being the third most common cause of UTI and the second most common cause of catheter-associated bacteriuria in long-term catheterized patients. P. mirabilis strains are able to form biofilm, a structure resistant to commonly used antibiotics [Maszewska et al, 2016]. Due to the production of biofilm on surfaces, treatment of P. mirabilis infections with common medications becomes difficult.…”

Use of polyvalent bacteriophages to combat biofilm ofProteus mirabiliscausing catheter-associated urinary tract infections

“…Instead, phage 65A caused a significant inhibition of biofilm formation and eradication of mature biofilms in 18 out of 50 tested strains and was characterized by strong lytic activity demonstrated by the spot test (Maszewska et al . ). It was chosen as a third cocktail component.…”

“…mirabilis ‐specific phages presenting different host ranges (Table ), isolated from urban wastewater from the Group Sewage Treatment Plant in Lodz and the Station of Water Supply and Sewage in Zgierz by the modified enrichment technique (Maszewska et al . ). The phage samples were stored at 4°C.…”

“… The adopted evaluation system of bacterial lawn lysis degree on spot when determining the host range using 50 Proteus mirabilis strains: 4—complete bacterial lysis on the spot, 3—very weak bacterial growth on the spot, 2—weak bacterial lysis on the spot, 1—very weak bacterial lysis on the spot (Maszewska et al . ). Phages selected as cocktail components in this study are highlighted in bold. …”

“…The aim of this paper was to evaluate the ability of previously isolated bacteriophages (Maszewska et al . ) and developed phage cocktail to combat biofilm of different uropathogenic P. mirabilis strains and to characterize phages with the strongest antibiofilm activity.…”