Eradication of biofilms that may harbor pathogens in
water distribution
systems is an elusive goal due to limited penetration of residual
disinfectants. Here, we explore the use of engineered filamentous
coliphage M13 for enhanced biofilm affinity and precise delivery of
lytic polyvalent phages (i.e., broad-host-range phages lysing multiple
host strains after infection). To promote biofilm attachment, we modified
the M13 major coat protein (pVIII) by inserting a peptide sequence
with high affinity for Pseudomonas aeruginosa (P. aeruginosa) extracellular
polysaccharides (commonly present on the surface of biofilms in natural
and engineered systems). Additionally, we engineered the M13 tail
fiber protein (pIII) to contain a peptide sequence capable of binding
a specific polyvalent lytic phage. The modified M13 had 102- and 5-fold
higher affinity for P. aeruginosa-dominated
mixed-species biofilms than wildtype M13 and unconjugated polyvalent
phage, respectively. When applied to a simulated water distribution
system, the resulting phage conjugates achieved targeted phage delivery
to the biofilm and were more effective than polyvalent phages alone
in reducing live bacterial biomass (84 vs 34%) and biofilm surface
coverage (81 vs 22%). Biofilm regrowth was also mitigated as high
phage concentrations induced residual bacteria to downregulate genes
associated with quorum sensing and extracellular polymeric substance
secretion. Overall, we demonstrate that engineered M13 can enable
more accurate delivery of polyvalent phages to biofilms in flow-through
systems for enhanced biofilm control.