Karla Veloso Gonçalves Ribeiro
1,2, Cleberson Ribeiro
2, Roberto Sousa Dias
2, Silvia Almeida Cardoso
3, Sergio Oliveira de Paula
2, Jose Cola Zanuncio
4, Leandro Licursi de Oliveira
2*1 Núcleo de Microscopia e Microanálise, Universidade Federal de Viçosa, 36570-900, Viçosa, Minas Gerais, Brasil.
2 Departamento de Biologia Geral, Universidade Federal de Viçosa, 36570-900, Viçosa, Minas Gerais, Brasil.
3 Departamento de Medicina e Enfermagem, Universidade Federal de Viçosa, 36570-900, Viçosa, Minas Gerais, Brasil.
4 Departamento de Entomologia, Universidade Federal de Viçosa, 36570-900, Viçosa, Minas Gerais, Brasil.
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.