In 2000, we surveyed microbiologists in 388 clinical laboratories, which tested an estimated 339,000 stool specimens in 1999, about laboratory methods and policies for the routine testing of stool specimens for Salmonella, Shigella, Campylobacter, and Vibrio species, Yersinia entercolitica, and Escherichia coli O157:H7. The results were compared with those of similar surveys conducted in 1995 and 1997. Although these laboratories reported routinely testing for Salmonella, Shigella, and Campylobacter species, only 57% routinely tested for E. coli O157:H7, 50% for Y. entercolitica, and 50% for Vibrio species. The mean proportions of stool specimens that yielded these pathogens were as follows: Campylobacter, 1.3% of specimens; Salmonella, 0.9%; Shigella, 0.4%; and E. coli O157:H7, 0.3%. The proportion of laboratories that routinely tested for E. coli O157:H7 increased from 59% in 1995 to 68% in 2000; however, the proportion of stool specimens tested decreased from 53% to 46%. E. coli O157:H7 should be routinely sought in stool specimens submitted for microbiologic culture.