Thermal processing methods such as pasteurization and sterilization are the most common practices to ensure the microbial safety of food. At the same time, these methods can cause significant changes in food related to both quality deterioration and reduced nutritional value. The increasing demand of consumers for foods free from microbiological load, and with high nutritional value has turned the interest of the scientific community and the food industry to the adoption of non-thermal techniques. Ozone and plasma treatments are among the non-thermal ones and they are emerging and promising techniques as they offer high microbial safety and extended self-life. Many studies have also been conducted in recent years with the effect of these methods on food bioactives with very encouraging results. It has been found that in most cases, these techniques can increase the content of many food bioactives in various foods, both raw and processed and mainly in foods of plant origin. The effect of these techniques on food bioactives can be different for each food and it depends on many factors with the most important being the food matrix, the time of application, the ozone concentration and plasma characteristics. This chapter evaluates the impact of ozone and plasma application on food bioactives.