Colonization of diatoms in and on porcine bone substrate and considerations of diatom ecology for forensic science.

The presence of diatom algae in bone marrow has been used as forensic evidence of drowning for several decades; however, these studies are based on known or suspected recent drowning events. This study addresses the potential for diatoms to enter the bone marrow of skeletal remains, that is, de-fleshed long bones post-mortem. In laboratory and field experiments, bones were either inflicted with two access points by a cut and acid pitting or left intact. The bones were submerged in water for at least 1 week and up to 3 months. Samples of the bone surface and marrow were inspected for diatoms. The analysis considered the time required for diatoms to enter marrow and whether genus characteristics like size or mobility affect entry. The presence of an access point influenced diatom entry in that bones without an introduced access point had zero to one diatom present in the marrow, whereas a bone with an access point had >150 diatoms present in the marrow. The results of both laboratory and field phases suggest that diatoms will reliably colonize bone in as quickly as 1 week, establishing and maintaining communities for at least 3 months. However, the bone surface assemblages differ from the source community. Bone marrow displayed even more restrictive access to diatom colonization, resulting in communities dominated by small raphid diatoms. Based on these findings, we suggest some caveats on the use of diatoms as trace evidence in forensic science with recommendations for future avenues of research.

A method for bone marrow extraction of diatoms for forensic science applications.

Diatoms show potential as trace evidence indicators, particularly as evidence of drowning. Often, the diatom test to diagnose drowning is done on soft tissue or bone marrow from a recently deceased individual. This method presented here combines elements from previous forensic literature and methods of diatom isolation in phycology to extract diatoms from bone marrow of skeletal remains for forensic use. This diatom extraction method is time-efficient, minimizes contamination risk, and produces samples of intact diatoms. This method is designed to complete sample preparation within 24 h, sampling the bone for diatoms internally and externally. This method was developed using porcine long bones submerged in water with live diatoms for up to 3 months. Three marrow samples were extracted from each bone so the method was developed using 102 marrow samples. Additionally, 132 surficial bone and environmental samples were collected and prepared during method development. To briefly summarize the method, the bone joints were cut off with an angle grinder in a biosafety hood to expose the marrow, which was removed from the hip, knee, and shaft as separate samples. The marrow was digested with nitric acid at 400°C in glass beakers before being centrifuged with DI water, plated onto microscope slides, and observed with a compound microscope. Observation found good preservation of unbroken diatom cell walls throughout the process. This method can be used to prepare diatoms as forensic trace evidence.

Laurentian Great Lakes phytoplankton and their water quality characteristics, including a diatom-based model for paleoreconstruction of phosphorus.

Recent shifts in water quality and food web characteristics driven by anthropogenic impacts on the Laurentian Great Lakes warranted an examination of pelagic primary producers as tracers of environmental change. The distributions of the 263 common phytoplankton taxa were related to water quality variables to determine taxon-specific responses that may be useful in indicator models. A detailed checklist of taxa and their environmental optima are provided. Multivariate analyses indicated a strong relationship between total phosphorus (TP) and patterns in the diatom assemblages across the Great Lakes. Of the 118 common diatom taxa, 90 (76%) had a directional response along the TP gradient. We further evaluated a diatom-based transfer function for TP based on the weighted-average abundance of taxa, assuming unimodal distributions along the TP gradient. The r(2) between observed and inferred TP in the training dataset was 0.79. Substantial spatial and environmental autocorrelation within the training set of samples justified the need for further model validation. A randomization procedure indicated that the actual transfer function consistently performed better than functions based on reshuffled environmental data. Further, TP was minimally confounded by other environmental variables, as indicated by the relatively large amount of unique variance in the diatoms explained by TP. We demonstrated the effectiveness of the transfer function by hindcasting TP concentrations using fossil diatom assemblages in a Lake Superior sediment core. Passive, multivariate analysis of the fossil samples against the training set indicated that phosphorus was a strong determinant of historical diatom assemblages, verifying that the transfer function was suited to reconstruct past TP in Lake Superior. Collectively, these results showed that phytoplankton coefficients for water quality can be robust indicators of Great Lakes pelagic condition. The diatom-based transfer function can be used in lake management when retrospective data are needed for tracking long-term degradation, remediation and trajectories.