Biofilm is defined as a community in which microorganisms adhere to a living or inanimate surface, embedded in a gelatinous layer in a self-produced matrix of extra polymeric substances, adhered to each other, to a solid surface or to an interface. Adverse environmental conditions caused biofilm formation by inducing transition of microorganisms from planktonic cell form to sessile cell form and altered metabolism of bacteria in biofilms. Bacteria in biofilm matrix produce the specific secondary metabolites and gain robustness. Although biofilms are often accepted as potentially destructive for clinical and other industrial fields, many biofilms are beneficial and there are several reports related to the positive use of these biofilms. Beneficial biofilms could be used for wide applications (antibacterial, food fermentation, biofertilizer, filtration, biofouling, prevention of corrosion, antimicrobial agents, wastewater treatment, bioremediation and microbial fuel cells) in food, agricultural, medical, environment and other fields. According to previous reports, certain strains including Bacillus spp. (B. subtilis, B. thuringiensis, B. brevis, B. licheniformis, Bacillus polymyxa, Bacillus amyloliquefaciens) Lactobacillus spp. (L. casei, L. paracasei, L. acidophilus, L. plantarum, L. reuteri) Enterococcus spp. (E. casseliflavus, E. faecalis, E. faecium), Pseudomonas spp. (P. fluorescens, P. putida and P. chlororaphis), Acetobacter aceti, some fungi and Pseudoalteromonas sp., etc. led to beneficial biofilm formation. Food and agricultural industry may mostly benefit from biofilms in terms of their biochemical, fermentative, antimicrobial and biotechnological characteristics. Microorganisms in biofilm matrix could positively affect quality characteristics of food products such as texture, biochemical composition and sensorial properties via the production of specific secondary metabolites. Additionally, biofilms have an importance in water and soil safety of agricultural land. The present chapter highlights beneficial biofilm applications in food and agriculture industry.