Sanitation guidelines for animal research facilities state that disinfection is achieved by application of high-temperature water (143 to 180 °F [62 to 82 °C]) or detergents and disinfectants. However, these guidelines are based on requirements for pasteurization, which may be unnecessarily stringent for the sanitation of nonfood items and do not address the theoretical sanitation potential of water at temperatures below 143 °F (62 °C). Recent literature indicates that water temperatures below 143 °F (62 °C) can also provide effective sanitation. In this study, we compared cagewash cycles at low (100 °F [38 °C] and 120 °F [49 °C]) and high (standard) (180 °F [82 °C]) temperatures and evaluated sanitation efficacy by using ATP swabs and RODAC plates. Low-temperature loads were washed either with or without prior treatment of a chemical disinfectant (10% bleach). The 100 °F (38 °C) cycle was not sufficient for sanitization without bleach pretreatment. However, the 120 °F (49 °C) cycle effectively sanitized cages without bleach pretreatment. Validation of effective sanitation at a lower water temperature (120 °F [49 °C]) can improve cagewash logistics and reduce costs as compared with standard (180 °F [82 °C]) high-temperature cycles.