The increasing number of agricultural biogas plants and higher amounts of digestate spread on agricultural land arouse a considerable interest in the hygiene situation of digested products. This chapter reviews the current knowledge on sanitation during anaerobic digestion and the hygienic status of digestate concerning a multitude of pathogens potentially compromising the health of humans, animals and plants. Physical, chemical and biological parameters influencing the efficiency of sanitation in anaerobic digestion are considered. The degree of germ reduction depends particularly on the resistance of the pathogen of concern, the processing conditions, the feedstock composition and the diligence of the operation management. Most scientific studies facing sanitation in biogas plants have provided data ascertaining reduction of pathogens by the biogas process. Some pathogens, however, are able to persist virtually unaffected due to the ability to build resistant permanent forms. As compared to the feedstock, the sanitary status of the digestate is thus improved or in the worst case, the sanitary quality remains almost unchanged. According to this, the spreading of digestate on agricultural area in accordance to current rules and best practice recommendations is considered to impose no additional risk for the health of humans, animals and plants.