Validation and optimization of the diatom L/D ratio as a diagnostic marker for drowning

If a dead body is discovered in water, it nearly always raises the question about the cause of death, often associated with the persistent problem to differentiate between a drowning incident and post-mortem immersion. In numerous cases, a reliable confirmation of death by drowning is often only possible by a combination of diagnoses obtained from autopsy and additional investigations. As to the latter, the use of diatoms has been suggested (and debated) since decades. Based on the consideration that diatoms are present in almost every natural waterbody and are unavoidably incorporated when water is inhaled, their presence in the lung and other tissues can provide evidence of drowning. However, the traditional diatom test methods are still subject of controversial discussion and suspected of erroneous outcome, predominantly through contamination. A promising alternative to minimize the risk of erroneous outcome seems to be disclosed by the recently suggested MD-VF-Auto SEM technique. Especially the establishment of a new diagnostic marker (L/D ratio), which represents the factorial proportion between the diatom concentration in lung tissue and the drowning medium, allows for clearer distinction of drowning and post-mortal immersion and is largely robust to contamination. However, this highly elaborated technique requires specific devices which are frequently unavailable. We therefore developed a modified method of SEM-based diatom testing to enable the use on more routinely available equipment. Process steps such as digestion, filtration, and image acquisition were thoroughly broken down, optimized, and ultimately validated in five confirmed drowning cases. Taking certain limitations into consideration, L/D ratio analysis provided promising results, even in cases of advanced decomposition. We conclude that our modified protocol indeed opens a way for a broader use of the method in forensic drowning investigation.