Bacteriophage Isolated from Sewage Eliminates and Prevents the Establishment of Escherichia Coli Biofilm

Abstract

Purpose: Biofilm growth exerts a negative impact on industry and health, necessitating the development of strategies to control. The objective of this work was study the lytic activity of the phage isolated from the sewage network in the formation and degradation of Escherichia coli biofilms.Methods: E. coli cultures were incubated in 96-well polystyrene microplates under controlled conditions to evaluate the biofilm formation. The E. coli cultures and established biofilms were treated with the suspensions of the vB_EcoM-UFV017 (EcoM017) bacteriophage obtained from sewage for 24 hours. The E. coli bacterial density was measured using absorbance at 600 nm and the biofilms were measured by crystal violet staining. Polystyrene coupons were used as support for Scanning Electron Microscopy and Confocal Microscopy to evaluate biofilm formation.Results: The E. coli strains formed biofilms in polystyrene microplates after 48 hours’ incubation. The highest EcoM017 phage titer, in the prevention and degradation experiments, reduced the bacterial growth and the quantity of biofilm formed by E. coli in 90.0% and 87.5%, respectively. The minimum dose capable of reducing the biofilms of this bacterium was 101 PFU/mL after 24 hours. The preformed E. coli biofilm mass was reduced 79% post exposure to the phage in the degradation assay. Microscopic analysis confirmed the results obtained in the plates assays.Conclusion: The EcoM017 phage prevented biofilm formation and degraded the E. coli-established ones. The EcoM017 phage isolated from sewage can reduce bacterial attachment and lyse the E. coli associated biofilm cells, offering biotechnological potential applicability for this phage.

Bacteriophages inhibit biofilms formed by multi-drug resistant bacteria isolated from septic wounds

“…In principle under static conditions, the released progeny will attach to the neighboring bacteria within the biofilms, whereas in case of dynamic conditions, the progeny of virus can target and lyse the entire span of bacterial colonies within the biofilms because of agitation in the experimental setup (Azeredo and Sutherland, 2008; Sillankorva et al, 2010; Azeredo and Sillankorva, 2018). We observed that K. pneumoniae and E. coli showed a significantly higher number of cells as compared to P. aeruginosa and S. aureus under DR condition at 48 h of incubation; Under SR condition, significant growth of MDR-bacterial isolates exhibited successful colonization These observations are in correlation with previous studies (Hughes et al2010; Ribeiro et al, 2018). While in the case of single-species biofilm formed by S. aureus and treated with phage (vB_SAnS_SADP1), we noticed degraded biofilm with some single coccus spread on the coverslip.…”

“…The biofilms coated on glass slides were fixed with glutaraldehyde (2.5%) and dehydrated through a series of graded ethanol (30-100%) for 5 minutes. Further, the glass slides were sputtered with gold after critical point drying, and the aggregated biofilms were examined using Scanning electron microscopy (FEI, Tecnai G-2S Twin) (Ceri et al, 1999; Ribeiro et al, 2018).…”

“…Bacteriophages when delivered at the infection site can replicate to produce a large number of active progeny, which in turn produce elution factors and degrading enzymes that target the polysaccharide components of the host bacteria, leading to bacterial lysis (Ghannad and Mohammadi, 2012; Vieira et al, 2012; Nouraldin et al, 2016). Though many studies have shown bacteriophage mediated mechanism of biofilm degradation, very few studies have used biofilms formed by clinical isolates (Pires et al, 2011; Danis-Wlodarczyk et al, 2015; Deshpande Kaistha and Kaistha, 2017; Oliveira et al, 2017; Ribeiro et al, 2018). Hence, there is an urgent need to identify bacteriophages that can act on the biofilms formed by bacteria isolated from wounds.…”

Isolation and characterization of six gamma-irradiated bacteriophages specific for MRSA and VRSA isolated from skin infections

“…In this study, all the isolated bacteriophages were isolated from sewage Figure 2. Ribeiro et al (2018) could successfully isolate bacteriophages from sewage. They revealed that to the sewage rich contents of bacteria and organic matters.…”

Genomic, morphological and functional characterisation of novel bacteriophage FNU1 capable of disrupting Fusobacterium nucleatum biofilms

“…Bacteriophages lytic against other types of bacteria such as Pseudomonas aeruginosa 25 , Escherichia coli 26 and Streptococcus mutans 27 have been shown to be able to disrupt mono-biofilms formed by their respective hosts. The potential exists for use of bacteriophages against biofilms in periodontal infection 18,19 and colon cancer.…”