Susceptibility testing of anaerobes has been described for approximately 45 years, some 15 years fewer than descriptions for aerobic organisms. During that time period, >16 methods, >16 media, and a host of other variables have been described in the United States literature, culminating in the most recent standards published by the National Committee for Clinical Laboratory Standards (NCCLS) in 2001. These new guidelines include a single agar dilution reference method as well as an alternative minimal inhibitory concentration method validated by extensive multilaboratory collaborative trials. In addition, the Food and Drug Administration-approved E-test has proved valuable as a user-friendly alternative to NCCLS methods. Highlights of the "evolution" of various methods, their variations and their failures, factors that affected the development of the current standards, and the rationale for susceptibility testing of anaerobes are discussed herein.