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<p>Human decomposition is a little-understood process with even less currently known about the microbiology involved. The aim of this research was to investigate the bacterial community associated with exposed decomposing mammalian carcasses on soil and to determine whether changes in this community could potentially be used to determine time since death in forensic investigations. A variety of soil chemistry and molecular biology methods, including molecular profiling tools T-RFLP and DGGE were used to explore how and when bacterial communities change during the course of a decomposition event. General bacterial populations and more specific bacterial groups were examined. Decomposition was shown to cause significant and sequential changes in the bacterial communities within the soil, and changes in the bacterial community often correlated with visual changes in the stage of decomposition. Organisms derived from the cadavers and carcasses were able to be detected, using molecular methods, in the underlying soil throughout the decomposition period studied. There was little correlation found between decomposition stage and the presence and diversity within the specific bacterial groups investigated. Organisms contributing to the changes seen in the bacterial communities using molecular profiling methods were identified using a cloning and sequencing based technique and included soil and environment-derived bacteria, as well as carcass or cadaver-derived organisms. This research demonstrated that pig (Sus scrofa) carcass and human cadaver decomposition result in similar bacterial community changes in soil, confirming that pig carcasses are a good model for studying the microbiology of human decomposition. The inability to control for differences between donated human cadavers made understanding the human cadaver results difficult, whereas pig carcass study allowed many variables to be held constant while others were investigated. The information gained from this study about the bacteria associated with a cadaver and how the community alters over the course of decomposition may, in the future, enable the development of a forensic post mortem interval estimation tool based on these changes in the bacterial community over time. The findings in this thesis suggest that high variability between human bodies and their microflora is likely to present a challenge to the development of such a tool, but further study with emerging high-throughput molecular tools may enable identification of microbial biomarkers for this purpose.</p>

<p>Soil is frequently encountered as trace evidence in forensic science case-work, but because of the limitations of current analytical techniques, this evidence is rarely utilised. A technique has been developed that allows comparisons of soil samples to be made, based on molecular analysis of the bacterial communities living in the soil. This project assesses the practicality of using this technique, known as 16S rDNA T-RFLP community profiling, for forensic soil analysis, by refining the basic methodology and performing a preliminary evaluation of its reproducibility and utility. Initial difficulties associated with generating profiles from soil samples have been overcome through methodology improvement, and the technique has been found to be effective for generating simple, visual profiles that clearly demonstrate differences between soil samples. Soil bacterial community DNA profiling is likely to be a powerful yet simple forensic tool, providing the ability to routinely use soil as associative evidence. The potential for using the same technology to develop a time since death or post mortem interval (PMI) estimation tool was also investigated. This study monitored the changes in the soil bacterial community beneath decomposing human cadavers and pig carcasses and showed that community change is dynamic and progressive. These changes are caused by fluctuations in specific bacterial species populations that are able to utilise organic breakdown products released from the body over time. Release of the body’s natural microflora into the underlying soil may also contribute to an altered bacterial community. This project has demonstrated that the soil microbial community clearly changes over the course of decomposition, and potential exists for development of a PMI estimation tool based on soil bacterial community succession.</p>