Postmortem submersion interval (PMSI) estimation from the microbiome of sus scrofa bone in a freshwater lake

While many studies have developed microbial succession-based models for the prediction of postmortem interval (PMI) in terrestrial systems, similar well-replicated long-term decomposition studies are lacking for aquatic systems. Therefore, this study sought to identify temporal changes in bacterial community structure associated with porcine skeletal remains (n = 198) for an extended period in a fresh water lake. Every ca. 250 ADD, one cage, containing 5 ribs and 5 scapulae, was removed from the lake for a total of nineteen collections. Water was also sampled at each interval. Variable region 4 (V4) of 16S rDNA was amplified and sequenced for all collected samples using Illumina MiSeq FGx Sequencing platform; resulting data were analyzed with the mothur (v1.39.5) and R (v3.6.0). Bacterial communities associated with ribs differed significantly from those associated with scapulae. This difference was mainly attributed to Clostridia, Holophagae, and Spirochaete relative abundances. For each bone type, alpha-diversity increased with ADD; similarly, beta-diversity bacterial community structure changed significantly with ADD and were explained using environmental parameters and inferred functional pathways. Models developed using 24 rib and 34 scapula family-level taxa allowed the prediction of PMSI with root mean square error of 522.97 ADD (similar to 57 days) and 333.8 ADD (similar to 37 days), respectively.

Automated summary

This study investigated temporal changes in bacterial community structure associated with porcine skeletal remains in a freshwater lake, revealing significant differences between rib and scapulae-associated bacterial communities. Alpha-diversity increased with accumulated degree days (ADD) for each bone type, while beta-diversity bacterial community structure changed significantly with ADD, explained by environmental parameters and functional pathways. Models utilizing family-level taxa allowed for the prediction of postmortem submersion interval (PMSI) with reasonable accuracy.