Comparative soil bacterial metabarcoding after aboveground vs. subsurface decomposition of Mus musculus

Outdoor microcosms, metabarcoding with next-generation sequencing of the 16S rRNA bacterial gene, total body score (TBS) and physicochemical analyses were used to monitor Mus musculus decomposition aboveground (A) and in the subsurface (S), and compared to soil-only controls (C). As determined by MaAsLin2 analysis, significant shifts in bacterial communities at 30 cm depths within the A, S and C treatments distinguished control from experimental soils, and between aboveground and subsurface deposition, demonstrating the potential for gravesoil discrimination during the first 90 days. For example, Dokdonella (p = 0.0002), Edaphobaculum (p = 0.0004) and Lacibacter (p = 0.0034) recorded significant shifts relative to sampling time. Furthermore, Massilia (p = 0.0005), Mycobacterium (p = 0.0006) and Sandaracinus (p = 0.0007) increased in abundance for the aboveground mice treatments. This was confirmed with ANOSIM where p = 0.0082 showed statistically significant difference between the aboveground and subsurface deposition. TBS and physicochemical analyses suggested that nutrient release into the soils occurred during active decay and skin rapture on days 7-13 in the subsurface and days 13-20 aboveground, with a particular increase in soil potassium concentration on day 15. Significant differences in soil temperatures resulted between A and S vs. C microcosms, aligning with atmospheric temperature changes. In summary, complementary application of metabarcoding, total body score, exogenous and physicochemical methods for postmortem interval estimation and clandestine grave location highlighted the feasibility of using temperature records downloaded from meteorological stations and portable X-ray fluorescence as indicators for various phases of decomposition.