Vibrios are natural inhabitants of aquatic environments and form symbiotic or pathogenic relationships with eukaryotic hosts. Recent studies reveal that the ability of vibrios to form biofilms (i.e. matrix-enclosed, surface-associated communities) depends upon specific structural genes (flagella, pili and exopolysaccharide biosynthesis) and regulatory processes (two-component regulators, quorum sensing and c-di-GMP signaling). Here, we compare and contrast mechanisms and regulation of biofilm formation by Vibrio species, with a focus on Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio fischeri. Although many aspects are the same, others differ dramatically. Crucial questions that remain to be answered regarding the molecular underpinnings of Vibrio biofilm formation are also discussed.