Identifying the scene of a crime through pollen analysis.

The forensic analysis of pollen involves the comparison of crime scene and reference pollen samples. Successful matches are frequently used to solve time- or location-related crimes. Despite its prospects in criminal investigation, forensic palynology is still underused in casework due to inherent shortcomings such as its limited evidential weighting, scarcity of skilled palynologists dedicated to forensic casework and the laborious nature of analytical procedures. To address these challenges, the current state-of-the-art in forensic palynology is transiting from the traditional light microscopic methods that dominated the early days of palynology to more contemporary approaches like Raman spectroscopy, stable isotope analysis and DNA metabarcoding. The major challenges of these methods, however, include a lack of optimisation to forensic expectations and the unavailability of robust databases to permit accurate data interpretation, and quests to resolve these problems constitute the theme of current research. While reiterating the usefulness of pollen analysis in criminal investigation, this report recommends orthogonal testing as a way of improving the evidential weighting of forensic palynology.

Drone aerial imagery for the simulation of a neonate burial based on the geoforensic search strategy (GSS).

A woman reporting the homicide and burial of an infant in 2004 prompted the creation of an experimental simulated neonate grave shortly before the real search commenced. The real case, documented here, did not use aerial imagery, but used ground-penetrating radar (calibrated to the test site described here) to identify two locations that were probed for gas release and the deployment of victim recovery dogs. We suggest technological advances in remotely sensed aerial imagery that have developed since 2004 will demonstrate their use in focusing such searches by informing a Geoforensic Search Strategy (GSS) and suggesting locations accessible by a perpetrator to identify a burial location using the still-existent analogue site. To test this, in the spring of 2020 a DJI Mavic Pro drone was flown over the control site containing the simulated 2004 burial. Aerial image processing included the creation of orthomosaics, Normalised Difference Vegetation Index (NDVI), Visual Atmospheric Resistance Index (VARI), and photogrammetry. Conventional ground-based geophysical surveys using ground-penetrating radar, guided by this new type of information integrated into the GSS, confirmed that anomalies seen in drone data were the 16-year-old burial. We test this strategy using both the original simulated burial in Northern Ireland and further evaluate it in two recent simulated graves in the United States in more complex scenarios, but with successful results.

Contribution of plant anatomy to forensic investigation: Tree bark morphology.

Plant science has been more and more utilized in forensic investigation, although its full potential is still to be reached. Plant macroremains are a powerful tool to link a body or other evidence back to a primary crime scene as they can provide detailed information about its previous ecological and geographic location. However, plant macroremains are often poorly preserved and difficult to identify, as diagnostic elements are seldom present within the assemblage occurring on the scene. Plant fragments most likely to be found are those exposed to the environment and resistant to degradation. The bark of woody plants meets these requirements but the possibility of its identification at species level from small fragments is not known. Starting from a real homicide case, where bark splinters were found on the victim, we aimed to assess the forensic potential of bark identification from small fragments like those likely to occur on a crime scene. Two identification keys were prepared for 16 common lowland tree species from Northern Italy; one key used all the available anatomical traits, the second only those from the outer bark. The second key was not able to discriminate some couples of species unambiguously, but could identify the bark fragments of the homicide as Robinia pseudoacacia, as confirmed from direct comparison with a reference sample. Bark fragments deserve to be included into the macroremains to be analyzed during an investigation, but small samples could easily lack diagnostic traits, and the building of a reference collection should be encouraged.

Massively parallel sequencing is unlocking the potential of environmental trace evidence.

Massively parallel sequencing (MPS) has revolutionised the field of genomics enabling substantial advances in human DNA profiling. Further, the advent of MPS now allows biological signatures to be obtained from complex DNA mixtures and trace amounts of low biomass samples. Environmental samples serve as ideal forms of contact trace evidence as detection at a scene can establish a link between a suspect, location and victim. Many studies have applied MPS technology to characterise the biodiversity within high biomass environmental samples (such as soil and water) to address questions related to ecology, conservation, climate change and human health. However, translation of these tools to forensic science remains in its infancy, due in part to the merging of traditional forensic ecology practices with unfamiliar DNA technologies and complex datasets. In addition, people and objects also carry low biomass environmental signals which have recently been shown to reflect a specific individual or location. The sensitivity, and reducing cost, of MPS is now unlocking the power of both high and low biomass environmental DNA (eDNA) samples as useful sources of genetic information in forensic science. This paper discusses the potential of eDNA to forensic science by reviewing the most explored applications that are leading the integration of this technology into the field. We introduce novel areas of forensic ecology that could also benefit from these tools with a focus on linking a suspect to a scene or establishing provenance of an unknown sample and discuss the current limitations and validation recommendations to achieve translation of eDNA into casework.

Enhanced analysis of weathered crude oils by gas chromatography-flame ionization detection, gas chromatography-mass spectrometry diagnostic ratios, and multivariate statistics.

Artificially weathered crude oil "spill" samples were matched to unweathered suspect "source" oils through a three-tiered approach as follows: Tier 1 gas chromatography-flame ionization detection (GC/FID), Tier 2 gas chromatography-mass spectrometry (GC/MS) diagnostic ratios, and Tier 3 multivariate statistics. This study served as proof of concept for a promising and new method of crude oil forensics that applies principal component analysis (PCA) and partial least squares discriminant analysis (PLSDA) in tandem with traditional forensic oil fingerprinting tools to confer additional confidence in challenging oil spill cases. In this study, weathering resulted in physical and chemical changes to the spilled oils, thereby decreasing the reliability of GC/FID and GC/MS diagnostic ratios in source attribution. The shortcomings of these traditional methods were overcome by applying multivariate statistical tools that enabled accurate characterization of the crude oil spill samples in an efficient and defensible manner.

Using museum pelt collections to generate pollen prints from high-risk regions: A new palynological forensic strategy for geolocation.

The use of pollen as a forensic tool for geolocation is a well-established practice worldwide in cases ranging from the provenance of drugs and other illicit materials to tracking the travel of individuals in criminal investigations. Here we propose a novel approach to generation of pollen databases that uses pollen vacuumed from mammal pelts collected historically from international areas that are now deemed too high risk to visit. We present the results of a study we conducted using mammal pelts collected from Mexico. This new investigative technique is important because, although it would seem that the ubiquitous and geo-specific nature of pollen would make pollen analysis among the most promising forensic tools for law enforcement and intelligence agencies, it is not the case. The process is notoriously slow because pollen identification is a tedious task requiring trained specialists (palynologists) who are few in number worldwide, and the reference materials necessary for geolocation usually are rare or absent, especially from regions of the world that are no longer safe to visit because of war or threat of terrorism. Current forensic palynological work is carried out by a few highly trained palynologists who require accurate databases of pollen distribution, especially from sensitive areas, to do their jobs accurately and efficiently. Our project shows the suitability of using the untapped museum pelt resources to support homeland security programs. This first palynological study using museum pelts yielded 133 different pollen and spore types, including 8 moss or fern families, 12 gymnosperm genera and 112 angiosperm species. We show that the palynological print from each region is statistically different with some important clustering, demonstrating the potential to use this technique for geolocation.

Forensic palynology and the search for geolocation: Factors for analysis and the Baby Doe case.

First used over 50 years ago, forensic palynology is an important tool for law enforcement agencies. In most countries that use forensic palynology, microscopic pollen grains and spores are traditionally used in criminal investigations to link suspects to crime scenes or items. While still underutilized in many parts of the world, forensic palynology is increasingly being used to determine the region of origin, or geolocation, for persons and items of interest. Drawing upon the experience of the authors using trace pollen and spores to geolocate forensic samples, the types, methods, and variables of this type of analysis are discussed and demonstrated using the Baby Doe case from Massachusetts, USA as a case study. This is not an exhaustive list and every forensic sample is unique so the methods and experience presented here are intended to be a guide for future forensic and anti-terrorism investigations as forensic palynology becomes more commonplace in law enforcement agencies around the world.

A systematic review and meta-analysis of the fully formed wisdom tooth as a radiological marker of adulthood.

Assessment of third molar development on dental radiograms is one of the most commonly used methods of forensic age estimation. Despite widespread use and numerous studies, there is a paucity of aggregated data on how well a fully mature third molar identifies adulthood (> 18 years), the most important threshold in a medicolegal context. Therefore, a systematic review and meta-analysis was conducted. A total of 1229 studies were screened, and 82 studies were assessed for inclusion. Twenty-four studies, with a pooled cohort of 19,690 individuals, presented true positive (TP), true negative (TN), false positive (FP), and false negative (FN) outcomes, enabling meta-analysis. The outcomes were based on using the third molar in the fully mature stage as a diagnostic test to indicate age 18 years or above. The false positive rate (fall-out/1-specificity) was 3.1% (95% CI 2.1-4.6%), and the true positive rate (recall/sensitivity) was 51% (95% CI 44-58%). Diagnostic accuracy was 71%. These findings, while reassuring in terms of the low false positive rate, highlight the need for complementary age estimation methods to avoid a significant number of false negatives.

Memory at the Sharp End: The Costs of Remembering With Others in Forensic Contexts.

In many applied contexts where accurate and reliable information informs operational decision-making, emergency response resource allocation, efficient investigation, judicial process, and, ultimately, the delivery of justice, the costs of unfettered conversational remembering can be high. To date, research has demonstrated that conversations between co-witnesses in the immediate aftermath of witnessed events and co-witness retellings of witnessed events often impair both the quality and quantity of information reported subsequently. Given the largely negative impact of conversational remembering on the recall of both individual witnesses and groups of witnesses in this context, this review explores the reasons why these costs occur, the conditions under which costs are exacerbated, and how, in practical terms, the costs can be reduced in order to maximize the accuracy and completeness of witness accounts.

Fusion Bead Procedure for Nuclear Forensics Employing Synthetic Enstatite to Dissolve Uraniferous and Other Challenging Materials Prior to Laser Ablation Inductively Coupled Plasma Mass Spectrometry.

There is an increasing demand for rapid and effective analytical tools to support nuclear forensic investigations of seized or suspect materials. Some methods are simply adapted from other scientific disciplines and can effectively be used to rapidly prepare complex materials for subsequent analysis. A novel sample fusion method is developed, tested, and validated to produce homogeneous, flux-free glass beads of geochemical reference materials (GRMs), uranium ores, and uranium ore concentrates (UOC) prior to the analysis of 14 rare earth elements (REE) via laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). The novelty of the procedure is the production of glass beads using 9 parts high purity synthetic enstatite (MgSiO3) as the glass former with 1 part of sample (sample mass ∼1.5 mg). The beads are rapidly prepared (∼10 min overall time) by fusing the blended mixture on an iridium strip resistance heater in an argon-purged chamber. Many elements can be measured in the glass bead, but the rare earth group in particular is a valuable series in nuclear forensic studies and is well-determined using LA-ICP-MS. The REE data obtained from the GRMs, presented as chondrite normalized patterns, are in very good agreement with consensus patterns. The UOCs have comparable patterns to solution ICP-MS methods and published data. The attractions of the current development are its conservation of sample, speed of preparation, and suitability for microbeam analysis, all of which are favorable for nuclear forensics practitioners and geochemists requiring REE patterns from scarce or valuable samples.

Spectrophotometric determination of gold(III) in forensic and pharmaceutical samples and results complemented with ICP AES and EDXRF analysis.

Spectrophotometric method with three systems were developed here for the determination of gold(III) using o-dianisidine, aniline sulphate and catechol. Gold(III),in the system 1 it oxidizes o-dianisidine, in the system 2 it oxidizes catechol followed by its coupling with o-dianisidine, in the system 3 it oxidizes catechol followed by its coupling with aniline sulphate forming dye products with respective λmax 446nm, 540nm, and 505nm. All the three systems were optimized and analytical parameters were calculated. The molar absorptivity values were 9.27×104, 1.97×104 and 1.62×104 respectively for the systems 1, 2 and 3 with the corresponding Sandell sensitivity values (µgcm-2), 0.0021, 0.0096 and 0.011. The optimized systems were used for the determination of gold present in some forensic jewellery and pharmaceutical samples and the results obtained were compared with the results of all samples determined by Inductively Coupled Plasma - Atomic Emission Spectrometric method and a few of them were also complemented by Energy Dispersive X-Ray Fluorescent spectral analysis.

Voltammetric analysis of Pinus needles with physiological, phylogenetic, and forensic applications.

Polyphenolic compounds are electrochemically active components of vegetal matter which were targeted under simple experimental conditions to produce voltammetric profiles characterizing the metabolite composition. Application to bivariate and multivariate chemometric techniques permits to discriminate the species and age of plant leaves, illustrated here for the case of six Pinus species from two different subgenera. Such responses, associated with the electrochemical oxidation of polyphenolic compounds (quercetin, gallic acid, ellagic acid, among others), define a voltammetric profile which varies systematically with the age of the leaves for the different species. The application of this methodology for phylogenetic studies, plant physiology, forensic science, and chemoecology is discussed. Graphical Abstract Image of Pinus in a typical Mediterranean forest; Courtesy of the Botanic Garden of the University of Valencia.

Review and future prospects for DNA barcoding methods in forensic palynology.

Pollen can be a critical forensic marker in cases where determining geographic origin is important, including investigative leads, missing persons cases, and intelligence applications. However, its use has previously been limited by the need for a high level of specialization by expert palynologists, slow speeds of identification, and relatively poor taxonomic resolution (typically to the plant family or genus level). By contrast, identification of pollen through DNA barcoding has the potential to overcome all three of these limitations, and it may seem surprising that the method has not been widely implemented. Despite what might seem a straightforward application of DNA barcoding to pollen, there are technical issues that have delayed progress. However, recent developments of standard methods for DNA barcoding of pollen, along with improvements in high-throughput sequencing technology, have overcome most of these technical issues. Based on these recent methodological developments in pollen DNA barcoding, we believe that now is the time to start applying these techniques in forensic palynology. In this article, we discuss the potential for these methods, and outline directions for future research to further improve on the technology and increase its applicability to a broader range of situations.

A Rapid and Efficient Method for Evaluation of Suspect Testimony: Palynological Scanning.

A rapid method for evaluating suspect testimony is valuable at any stage in an inquiry and can result in a change of direction in an investigation. Rape cases, in particular, can present problems where a defendant renders DNA analysis redundant by claiming that the claimant consented to have sexual relations. Forensic palynology is valuable in confirming or eliminating locations as being crime scenes, thus checking the testimony of both parties. In contrast to some forensic disciplines, forensic palynology can provide critical information without time-consuming full analysis. Two cases are described where the palynological assemblages from comparator samples of pertinent places were compared with those obtained from clothing of claimants and defendants. The results of rapid microscopical scanning of relevant preparations led to early confessions, thus obviating the need for costly analyses and protracted court proceedings. A third case demonstrates the unbiased nature of this technique where a man, although innocent of any offense, lied about having visited the crime scene for fear of prosecution. This highlights the need for sensitive policing in claims of rape.

The spatial and temporal distribution of pollen in a room: forensic implications.

This paper presents two experimental studies that deal with the spatial and temporal distribution of pollen grains within a room of a domestic dwelling. The findings concur with the preliminary work of Morgan et al. [1] and provide greater detail as to the behaviour of pollen grains within indoor locations that are pertinent for forensic investigations. The spatial distribution of pollen in a room exhibits strong distance decay trends, with the majority of pollen recovered within 0.8m of its source. The pollen was found to persist in increasing quantities during the time the flowers were in the room. This study also shows that 20days after the flowers were removed, 25-32% of the original pollen was still present within the room. The influence of disturbance was investigated and whilst areas of high disturbance were found to retain less pollen than undisturbed locations, the influence of the proximity to source was a more dominant factor. These findings have significant implications for forensic investigation protocols, particularly the collection and interpretation phases of trace evidence analysis. The distribution of pollen around a room ensures that viable sources of trace pollen are available for transfer if contact is made between a location in the room and a suspect. The persistence of pollen many days after the flowers have been removed from a room indicates that many rooms in domestic dwellings will have distinctive assemblages that reflect the history of the flowers that have been displayed within that room in the past, and that these assemblages will persist and therefore be available for transfer. These preliminary findings indicate that investigation by forensic palynology in indoor domestic settings may well be an underutilised technique that has the potential to provide accurate and valuable intelligence and evidence for forensic enquiry.

Forensic identification of spilled biodiesel and its blends with petroleum oil based on fingerprinting information.

A case study is presented for the forensic identification of several spilled biodiesels and its blends with petroleum oil using integrated forensic oil fingerprinting techniques. The integrated fingerprinting techniques combined SPE with GC/MS for obtaining individual petroleum hydrocarbons (aliphatic hydrocarbons, polyaromatic hydrocarbons and their alkylated derivatives and biomarkers), and biodiesel hydrocarbons (fatty acid methyl esters, free fatty acids, glycerol, monoacylglycerides, and free sterols). HPLC equipped with evaporative scattering laser detector was also used for identifying the compounds that conventional GC/MS could not finish. The three environmental samples (E1, E2, and E3) and one suspected source sample (S2) were dominant with vegetable oil with high acid values and low concentration of fatty acid methyl ester. The suspected source sample S2 was responsible for the three spilled samples although E1 was slightly contaminated by petroleum oil with light hydrocarbons. The suspected source sample S1 exhibited with the high content of glycerol, low content of glycerides, and high polarity, indicating its difference from the other samples. These samples may be the separated byproducts in producing biodiesel. Canola oil source is the most possible feedstock for the three environmental samples and the suspected source sample S2.

Maximizing the usefulness of hypnosis in forensic investigative settings.

This is an article written for mental health professionals interested in using investigative hypnosis with law enforcement agencies in the effort to enhance the memory of witnesses and victims. Discussion focuses on how to work with law enforcement agencies so as to control for factors that can interfere with recall. Specifics include what police need to know about how to conduct case review, to prepare interviewees, to conduct interviews, and what to do with the results. Case examples are used to illustrate applications of this guidance in actual investigations.

A case of a false confession after an inadvertent hypnotic induction.

This article presents the case of a highly hypnotizable 28-year-old man who gave a false confession under police interrogation following an unintended hypnotic induction, but who was exonerated years later on the basis of DNA evidence. The author proposes that assessment of possible high hypnotizability be conducted in instances of otherwise unsubstantiated confessions.

Analytical Raman spectroscopic discrimination between yellow pigments of the Renaissance.

The Renaissance represented a major advance in painting techniques, subject matter, artistic style and the use of pigments and pigment mixtures. However, most pigments in general use were still mineral-based as most organic dyes were believed to be fugitive; the historical study of artists' palettes and recipes has assumed importance for the attribution of art works to the Renaissance period. Although the application of diagnostic elemental and molecular spectroscopic techniques play vital and complementary roles in the analysis of art works, elemental techniques alone cannot definitively provide the data needed for pigment identification. The advantages and limitations of Raman spectroscopy for the definitive diagnostic characterisation of yellow pigments that were in use during the Renaissance is demonstrated here in consideration of heavy metal oxides and sulphides; these data will be compared with those obtained from analyses of synthetic yellow pigments that were available during the eighteenth and nineteenth Centuries which could have been used in unrecorded restorations of Renaissance paintings.