Proper preservation of nutrient-rich raw food from microbial contamination is a major challenge faced by the food industry. The use of antibiotics and chemicals to address the issue of contamination in fresh produce and meat has led to antibiotic-resistant pathogens. In the current study, the anti-biofilm effect of bio-oil aqueous phase (BOAP) collected during pyrolysis of seaweed was evaluated against some major food-borne pathogens for its biofilm inhibition and eradication potential in mono- and dual-species biofilms. The results showed that BOAP could eradicate almost 75% of dual-species biofilms. Further, the preservative effect of the BOAP was examined using appropriate food samples (pineapple and prawns). There was profound reduction in the bacterial load in the food samples which shows that BOAP has effective food preservation property. SEM analysis also revealed the drastic reduction in biofilm in BOAP treated food samples. In future, natural preservatives and BOAP can be combined to increase shelf-life of the above food samples.