Vibrio bacteria naturally inhabit coastal waters and are frequently isolated from a variety of raw seafood, particularly oysters. Detection of Vibrio species in seafood and growing water has traditionally been problematic because of the limitations of conventional techniques based on plate culture methods. Moreover, no culture method is able to discriminate pathogenic populations. In order to provide education and training to individuals involved in seafood testing, a hands-on training course has been developed and offered through cooperative efforts of University of Maryland Extension (UME) and University of Maryland’s Joint Institute of Food Safety and Applied Nutrition (JIFSAN). The methods described in this manual are designed for detection and enumeration V. parahaemolyticus, V. vulnificus, and V. cholerae in oysters. The analytical methods are combination of procedures from the Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM) and other sources, and have been used as training protocols for the hands-on training course on Vibrio Detection.