Biofilms composed of anaerobic bacteria often result in persistent infections and chronic inflammation. Host immune cells have difficulties in clearing biofilm-related infections and this can result in unnecessary tissue damage due to the overproduction of proinflammatory mediators. The objective of this project was to generate mixed-species anaerobic biofilms composed of two opportunistic pathogens, Fusobacterium necrophorum and Porphyromonas levii and measure neutrophil responses. Neutrophils are a vital component of the innate immune system and the results show that neutrophils exposed to mixed-species planktonic bacteria exhibit a more extensive oxidative response than neutrophils exposed to biofilms composed of the same bacteria. The limited neutrophil response to biofilm bacteria may explain the reduced ability of the innate immune system to eradicate biofilm-associated infections. The results demonstrate that bacterial lipopolysaccharide plays a significant role in the stimulation of neutrophils, however the evidence points to the presence of other stimulatory molecules in the bacterial media.