This chapter discusses recent biosensor developments for the monitoring of food spoilage using polydiacetylene (PDA) and liposome-based systems. Consequently, PDA is gaining attention as a colorimetric indicator of food spoilage throughout processing, distribution, and consumption. Typical food contaminants include bacteria, pesticides, allergens, and toxins. The potential applicability of PDA biosensors as contaminant detection devices and freshness indicators has been broadly documented. E. coli is a common foodborne pathogen which significantly affects human health and the food industry. Illnesses caused by E. coli generally consist of a range of conditions including diarrhea, abdominal pain, and headaches, affecting children under 5 years old and older adults. In addition to E. coli, Salmonella is another common foodborne pathogen with typical sources ranging from poultry products to vegetables and generally affects the elderly and children.