BACKGROUND After cryptosporidiosis was reported in three workers caring for preweaned calves at an academic research laboratory, we sought to identify cases, determine risk factors, and implement control measures. METHODS A cryptosporidiosis case was defined as diarrhea duration ≥72 hr, abdominal cramps, or vomiting in an animal research laboratory worker during July 14-July 31. A confirmed case had laboratory evidence of Cryptosporidium infection. Staff were interviewed regarding illness, potential exposures, training, and personal protective equipment (PPE) standard operating procedures (SOPs). RESULTS The cryptosporidiosis attack rate (AR) was 74% (20/27); five were laboratory-confirmed. Median job training was 2 hr including respiratory-fit testing. No SOPs existed for doffing PPE. AR for workers who removed their gloves first was 84% (16/19) compared with 20% (1/5) for workers who removed gloves last (risk ratio = 4.2; P < 0.02). CONCLUSIONS This outbreak highlights the importance of adequate training, enforced proper PPE procedures, and promoting a culture of safety. Am. J. Ind. Med. 60:208-214, 2017. © 2017 Wiley Periodicals, Inc.