Tissue preservation in extreme temperatures for rapid response to military deaths

Many deployable forensic capabilities, including those used by the Australian Defense Force (ADF), employ mobile battery-operated fridge/freezers for DNA sample preservation that are not suitable for rapid response application due to their size and weight. These fridge/freezers are expensive, require regular specialised maintenance, and have a set payload. A variety of transport media are successful preservatives for DNA samples, however, there is no research specifically targeted to their suitability for operational environments where temperatures exceed 50 °C. This research examined whether sodium chloride (NaCl), ethanol, and dimethyl sulfoxide (DMSO) could preserve muscle and bone samples (fresh and early decomposition) as effectively as refrigeration, when stored at 21 °C, 45 °C, 55 °C, and 65 °C for at least one week. A total of 78 muscle and 78 bone samples were collected from an unknown deceased individual. Half of each tissue type was stored at 30 °C for 48 h to induce early decomposition. Following this, samples were stored in the transport media for one week at the above temperatures, and a control set of samples were refrigerated (-4 °C) without any transport media. Preserved samples would need to provide DNA profiles comparable to the refrigerated samples for the transport media to be considered a successful replacement method. NaCl and 70% ethanol preserved muscle samples (fresh and decomposed) up to 65 °C, as well as 70% ethanol and 20% DMSO for fresh bone samples. These results were comparable with refrigeration and therefore, these preservatives could be used in rapid response operations by the military and for disaster victim identification. Conversely, under the conditions of this study, 20% DMSO and 70% ethanol failed to consistently produce full DNA profiles from decomposed bone, and NaCl performed poorly at preserving DNA from fresh and decomposed bone samples.