Preventing the Transmission of Murine Norovirus to Mice (Mus musculus) by Using Dry-heat Sterilization

Abstract

A critical component of an animal care biosecurity plan includes the sterilization of materials that come into direct contact with the animals. Dry-heat sterilization is gaining popularity in animal research facilities due to lower cost, less space utilization, no water usage, and the ability to sterilize water-sensitive materials. Currently, dry-heat sterilization ovens are validated against Bacillus atropheus spore strips with the assumption that a lack of sporulation is equivalent to successful sterilization. However, no published studies describe sterilization of rodent cages that contain relevant rodent pathogens by using this method. To determine if a dry-heat sterilizer can sterilize rodent cages and bedding against relevant rodent pathogens, we created murine norovirus (MNV)-contaminated cages by using mice with known MNV infection and shedding. The contaminated cages were either sterilized with the dry-heat sterilizer or not sterilized. Naïve, 4-wk-old, CD-1 mice were placed in the dry-heat–sterilized cages, contaminated unsterilized cages, or standard autoclaved cages for 2 wk. The mice were subsequently placed into clean, autoclaved cages for the remainder of the study. Fresh fecal pellets were collected at weeks 0, 12, and 16 and submitted for MNV PCR. Whole blood was collected for MNV serology at weeks 0, 8, 12, and 16. At week 16, all mice that had been in the unsterilized contaminated cages were positive for MNV by both fecal PCR and serology, whereas the mice in the dry-heat–sterilized and autoclaved cages were negative for MNV by both methods at all time points. Our study supports the use of dry heat sterilization as a viable sterilization method for rodent cages and bedding.