Forensic Sci., Volume 6, Issue 1 (March 2026) – 31 articles

Background/Objectives: The opioid crisis is an ongoing problem in the United States, and fentanyl analogs play a major role in the issue, as novel fentanyl analogs are constantly being developed. Substitutions and additions to the fentanyl scaffold impact the potency of the substances and can sometimes influence the biotransformation of the drugs. This study aimed to determine whether one or more common metabolites could be detected among a group of five methoxylated fentanyl analogs, for the purpose of eventually providing a more rapid detection method for new and unknown fentanyl-related compounds in toxicological samples. Methods: In vitro metabolism via pooled human liver microsomes (HLMs) was performed for five methoxylated fentanyl analogs (para-methoxyfentanyl, ortho- and para-methoxy butyryl fentanyl, and ortho- and para-methoxy furanyl fentanyl) to generate metabolites. Assays were analyzed via liquid chromatography–tandem mass spectrometry. Results: Nine metabolites were detected. A common metabolite was observed between fentanyl analogs that were methoxylated at the para-position. Conclusions: Similarities between metabolites of five methoxylated fentanyl analogs were noted. It appeared that the major pathway of metabolism for methoxylated fentanyl analogs is largely consistent, regardless of the location of the methoxy substituent so long as the piperidine nitrogen is unobstructed and is available for N-dealkylation. This research provides valuable insight to potentially find new ways for toxicologists to identify novel methoxylated fentanyl analogs in biological matrices. Full article

Background/Objectives: Calcaneal spurs are pathological bone formations at entheseal attachment sites with clinical implications but limited forensic anthropological applications. While entheseal changes have been proposed as age estimation markers in forensic contexts, empirical validation remains insufficient, particularly for Southeast Asian populations. This study evaluated calcaneal spur utility for forensic age estimation in Thai skeletal remains while establishing population-specific osteological reference data for forensic individuation. Materials and Methods: The 3516 dry calcanei from 1758 Northeastern Thai skeletons (1031 males, 727 females; age 22–106 years) were examined. Spurs were classified by anatomical location as dorsal (D-type), plantar (P-type), or combined plantar–dorsal (P–D type). The morphometric measurements were performed bilaterally. Age-associated patterns were analyzed across four age cohorts (≤40, 41–50, 51–60, ≥61 years), and Random Forest machine learning classification tested forensic age estimation capacity using 10-fold cross-validation. Results: Overall prevalence reached 67.63% with distinctive P–D type predominance. While age-stratified prevalence increased from 24.56% (≤40 years) to 74.77% (≥61 years), Random Forest modeling explicitly demonstrated overall classification accuracy of 62.5%. Compared between sexes, the maximum length of calcaneal spurs was significantly longer in males. Dimensional analyses revealed weak age correlations and substantial inter-individual morphological variation precluded reliable age prediction. Interestingly, the unique P–D type distribution pattern (77.5% among spur-bearing individuals) may serve as an auxiliary marker for Thai population affinity assessment in forensic contexts. Conclusions: This study established the first comprehensive Thai-specific osteological reference for calcaneal spurs, revealing distinctive plantar–dorsal type predominance valuable for forensic population affinity assessment and provided population-specific baseline data for forensic individuation. Full article

Background/Objectives: Abortion legislation represents a complex intersection of medical practice, ethical considerations, and legal frameworks that demonstrate significant legal heterogeneity across Europe. This study undertakes a comprehensive comparative assessment of the statutory schemes governing abortion across the European continent, examining gestational limits, exceptional circumstances, and regulatory requirements. Methods: A comparative legal analysis was conducted across 31 European jurisdictions. Primary legislative instruments were identified and authenticated through official governmental sources, parliamentary databases, and legal repositories to ensure analysis of current consolidated legislation. Data extraction focused on gestational limits, exceptional circumstances, procedural requirements, and constitutional provisions to categorize jurisdictions into regulatory models. Additionally, a structured literature search was performed in PubMed and Scopus (2015–2025) using the keywords “abortion,” “law,” and “Europe.” From 297 screened records, 30 articles were selected to contextualize legislative evolution and scholarly discourse. Results: The comparative analysis identified substantial heterogeneity in European abortion legislation, revealing four distinct regulatory models. Most jurisdictions establish a legal limit for elective abortion of approximately 12 weeks of gestation, with variations ranging from 10 weeks to 24 weeks. Exceptions to gestational limits are widely recognized for maternal life-threatening conditions, severe fetal anomalies and pregnancies resulting from sexual violence. Conclusions: European abortion legislation reflects persistent regulatory pluralism rather than convergence toward a unified model. While commonality exists regarding early gestational limits for elective abortion, significant variation remains in exceptional circumstances, procedural requirements, and underlying regulatory philosophies. This heterogeneity impacts healthcare provision, cross-border reproductive care, and medico-legal practice. The identified regulatory models illustrate diverse balances between reproductive autonomy and state interests. Future research should examine the practical consequences of these diverse schemes on health outcomes and cross-border patient mobility. Full article

Dental hard tissues, through their remarkable resistance to degradation, represent one of the most durable biological materials available for postmortem investigation. The preparation of undecalcified or ground sections allows microscopic visualization of enamel, dentin and cementum structures, which can preserve chronological, physiological, or environmental information. This review provides a comprehensive overview of the forensic applications of dental hard tissue ground sections, focusing on methodological principles, interpretive potential and practical constraints. The literature in forensic odontology highlights their relevance for age estimation through tooth cementum annulation, identification of neonatal and accentuated stress lines, and the assessment of thermal or chemical alterations. While these methods have proven scientific validity in anthropology and histology, their forensic implementation remains limited by heterogeneity in protocols and interpretative subjectivity. Standardization of preparation techniques, digital imaging, and integration with complementary analyses such as micro-CT or SEM could enhance the reliability and medico-legal relevance of this classical but underused approach. Full article

Background/Objectives: Viper envenomation in Italy is uncommon but carries significant clinical and forensic implications: an average of 257 bites per year are estimated, with only one fatality. This epidemiological context necessitates careful consideration of the standard of care and professional liability The article aims to outline the clinical and medico-legal dimensions of viper bites within both national and international contexts. Methods: We report the case of a 40-year-old woman bitten by a viper in an urban environment. At the initial emergency department visit the presentation was classified as Grade 0 (“dry bite”) according to Boels and, after 21 h, was discharged in good condition. Three days later, she returned with worsening symptoms and CT imaging revealed intrafascial and subcutaneous edema. The subsequent onset of complications prompted a criminal malpractice investigation. Therefore, we performed a PubMed search which yielded 125 records; after applying eligibility criteria, 33 articles were included, supplemented by manual reference checking for a total of 60 sources reviewed. Results: Comparison with the Australian model suggests the need for more standardized care pathways in Italy, while accounting for local toxicological and epidemiological specificities. Conclusions: This case and the accompanying literature analysis highlight that, even in low-incidence settings, structured patient communication, multidisciplinary management, collaboration with Poison Control Centers, and adherence to good clinical practice are crucial for patient safety and for mitigating medico-legal risk. Full article

Introduction: Gender-based violence and femicide represent the most extreme manifestation of a deep-rooted cultural distortion embedded within patriarchal social structures. In this study, adopting a comparative and multidisciplinary approach, we analyzed the evolution of international legislation and the major historical milestones in the protection of women’s rights and the prevention of gender-based violence at both the global and Italian levels. Specific protocols such as the “Pink code” were examined, with particular attention to medico-legal implications and the clinical management of victims, highlighting how violence against women continues to be fuelled by stereotypes, discrimination, and unequal power relations. Materials and Methods: Gender-based violence and femicide were examined from both national and international perspectives. A total of 73 scientific articles in English and 28 legal sources were selected from an initial pool of 918 publications, through a narrative review with a structured search strategy of international and Italian legislation and scientific literature. Electronic databases (PubMed and Google Scholar) were searched for the period 2000–2025. Only original observational studies, medico-legal analyses, epidemiological reports, and forensic case series were included. Cases primarily related to pregnancy, migration, infanticide, suicide, or substance abuse were excluded to reduce heterogeneity and focus on violence rooted in gender-based power asymmetries. Results: The legislative analysis shows a progressive strengthening of protection mechanisms, particularly between 2012 and 2023, following the ratification of the Istanbul Convention, the increase in intimate partner violence, and the COVID-19 pandemic. In Italy, the repeal of discriminatory norms and the introduction of specific legislative measures have led to increased attention toward prevention, protection, and prosecution of gender-based violence. Protocols such as the ‘Pink Code’, an Italian hospital-based multidisciplinary pathway activated mainly in emergency departments for the early identification, clinical care, medico-legal documentation, and judicial protection of victims of gender-based violence, have improved multidisciplinary management of victims within healthcare and judicial settings, although significant challenges remain regarding the full enforcement of legislation and the effective protection of women. The analysis focuses on female victims, in accordance with the Italian legal definition of gender-based violence, while other forms of gender-related violence were considered beyond the scope of this review. Conclusions: Despite substantial legal advances, combating gender-based violence clearly requires an integrated approach that combines prevention, assistance, and prosecution. Strengthening collaboration among institutions, healthcare services, and the judicial system—consistent with international recommendations—is essential to ensure an effective and rights-based response to victims. Overcoming the cultural and social barriers that perpetuate violence remains a fundamental priority, alongside promoting genuine gender equality. Full article

Background/Objectives: When analyzing human skeletal remains for human identification, the assessment of sexual dimorphism is fundamental because it underlies sex estimation, a key parameter of the biological profile, which reduces the number of candidates to approximately one half. Ideally, the most dimorphic bones (the pelvis and long bones) are used, but this analysis may be compromised when the skeletal remains are badly preserved, which is often the case in older skeletons. It is, therefore, necessary to investigate sexual dimorphism in alternative skeletal elements as the quantity and quality of bone structures present in ancient skeletons represent crucial aspects when assessing biological differences between the sexes. The present study aims to evaluate the degree of sexual dimorphism in selected skeletal elements as a preliminary step toward identifying sexually dimorphic parameters with potential applicability in future research on poorly preserved skeletons. Methods: A metric assessment of sexual dimorphism was performed on the clavicle, sternum, and seventh cervical vertebra from a 20th Century collection of identified skeletons from the Portuguese population from CESPU (CEIC), showing a variable degree of bone preservation. Results: Our preliminary data suggest that although all bones analyzed may have exhibited some degree of sexual dimorphism, five parameters—sternal body length, manubrium width, first stern-vertebrae width, clavicle maximum length, and cervical vertebral body height—showed the most promising results. Nevertheless, further studies are needed to validate these observations, using a bigger sample of badly preserved bones. Conclusions: These results represent a preliminary assessment of sexually dimorphic parameters, which may be of interest in circumstances where skeletal elements are poorly preserved. Full article

Background: Listeria monocytogenes is an ubiquitous foodborne pathogen capable of persisting in food-processing environments, domestic settings, and water systems; consequently, the mere co-detection of the microorganism in a patient and in a food product is insufficient to support criminal causation. In judicial proceedings, attribution of invasive listeriosis to a specific food and food business operator requires a pathogen-specific evidentiary chain that exceeds the standards used for public health surveillance. Methods: We performed a retrospective medico-legal and methodological analysis of a fatal listeriosis case that triggered criminal prosecution for foodborne homicide, systematically assessing the investigative chain according to temporal plausibility, food traceability, compliance with European and ISO microbiological standards, interpretation of whole-genome sequencing (WGS) data for an ubiquitous pathogen, and clinical causation of death. Results: The analysis identified critical weaknesses in the accusatory reconstruction, including incompatibility with established incubation periods, lack of proof that the sampled food lot corresponded to the product actually consumed, deviations from standard microbiological procedures, reliance on a poorly documented “clinical” isolate for WGS, non-specific genomic findings within a widely disseminated clonal complex, and the presence of plausible alternative environmental reservoirs. Clinically, the immediate cause of death was more consistent with hypovolemic shock due to uncontrolled gastrointestinal bleeding than with ongoing listerial sepsis. Conclusions: This article demonstrates how, in cases involving ubiquitous pathogens, failure to adopt a pathogen-specific investigative and interpretative framework may lead to methodologically fragile criminal allegations, potentially increasing litigation burden and costs for the justice system. A methodologically robust approach integrating microbiology, genomics, epidemiology and medico-legal causation analysis is essential for fair and scientifically sound criminal proceedings. Full article

Background: Stature estimation is a key parameter in biological profiling of human skeletal remains and thus in aiding in the identification process of the deceased. Various methods, including anatomical approaches, multifactorial regressions and organic correlation approaches, have been developed to estimate stature. Challenges arise from incomplete or altered remains, outdated reference samples, and the lack of population specific studies. This paper critically examines genetic influences and population-specific factors, the present status quo, recent developments and the challenges in stature estimation in forensic anthropology. Methods: The study appraises the current status, challenges, and future directions of stature estimation in forensic anthropology and bioarchaeology. The open-source literature is systematically identified, and relevant osteological and imaging-based studies are screened and prioritised when they report original empirical data or clearly defined methodological developments, enabling consistent extraction of sample and population descriptors, modelling strategies, and limitations. Included studies are then synthesised comparatively and summarised in a study table to support direct cross-study evaluation. Results: Stature estimation accuracy is shown to depend on population, ancestry, sex, time period, and environment, with cross-population or cross-temporal equation transfer identified as a key source of bias. Limitations include fragmented/altered remains and non-representative reference samples, while newer imaging and probabilistic approaches improve performance when supported by suitable data and explicit error quantification. Conclusions: In the present article, stature estimation is positioned as a context-dependent, evidence-weighted process rather than a fixed calculation. Full article

Background/Objectives: The Casablanca-Settat region of Morocco, located at the interface between Arab and Amazigh cultural zones, has only recently been investigated using autosomal short tandem repeat (STR) markers. The objective of this study was to characterize the genetic diversity and forensic efficiency of 15 autosomal STR loci in the Casablanca-Settat population and to evaluate its genetic relationships with other Moroccan populations. Methods: Fifteen autosomal STR loci were genotyped in 138 unrelated Arabic-speaking individuals from the Casablanca-Settat region. Allele frequencies, Hardy–Weinberg equilibrium, and standard forensic parameters were calculated. The genetic structure of the population was further examined through comparative analyses with 12 previously published Moroccan reference populations using multivariate and phylogenetic approaches. Results: A total of 146 distinct alleles were identified across the 15 loci. D18S51 was the most polymorphic marker (Ho = 0.9203), whereas D3S1358, TPOX, D5S818, and D16S539 exhibited lower allelic diversity. No statistically significant deviation from Hardy–Weinberg equilibrium was detected after correction for multiple testing. The combined power of discrimination exceeded 0.99, and the combined power of exclusion reached 0.99999965, demonstrating the high forensic efficiency of the STR panel. Population structure analyses positioned the Casablanca-Settat population within an intermediate genetic cluster, closely related to central Moroccan populations, consistent with historical gene flow and admixture. Conclusions: This study provides robust autosomal STR reference data for the Casablanca-Settat population, confirming the suitability of these markers for forensic identification in Morocco and offering valuable insights into regional population structure and genetic diversity. Full article

Background/Objectives: Personal identification is a cornerstone of Forensic Medicine. Historical records indicated that 27 Italian soldiers were executed by Tito’s army in April 1945 and buried in the mass grave of Ossero (Cres Island, Croatia). Methods: The remains, exhumed in 2019 by Croatian authorities and transferred to the Sacrario d’Oltremare (Bari, Italy), underwent radiographic and anthropological analyses. Genetic analysis was later performed on 147 bone samples, leading to the identification of 10 soldiers. Results: Anthropological analyses revealed commingled remains of at least 34 individuals (15–45 years; 161–181 cm), including eight skeletal elements of female sex. Forensic examination confirmed cranial and post-cranial gunshot wounds. Conclusions: Extensive commingling and fragmentation of the remains prevented full reconstruction of individual skeletons, yet comparison with ante-mortem data supported their identification as the aforementioned soldiers. Unexpected findings, including historically undocumented females, were confirmed by DNA analysis. These findings underscore the value of a multidisciplinary approach to optimize recovery and subsequent forensic and genetic investigations. Full article

Background/Objectives: DNA profiling in forensic investigation typically compares genetic profiles, usually derived from the analysis of STR markers. However, this method has limitations when there is no biological reference sample or match in the DNA database. The aim of the current study is thus to replicate, in an Italian cohort, epigenetic markers previously identified in the literature for distinguishing tobacco smokers from non-smokers or estimating chronological age, so as to help narrow down the pool of suspects. Methods: DNA methylation at four CpG dinucleotides located around the cg05575921 site of the AHRR gene, widely associated with tobacco consumption, was measured. Additionally, five CpG dinucleotides in the ELOVL2, FHL2, KLF14, TRIM59, and C1orf132 genes were examined for chronological age estimation in buccal swab samples of 102 volunteers through pyrosequencing. Results: A multiple linear regression model for estimating chronological age shows that ELOVL2-C7, C1orf132-C1, and TRIM59-C7 have a significant effect on age. In this model, the prediction error increases with age. Two logistic regression models were used for determining smoker/non-smoker status, proving that two CpG sites significantly influence the odds of being classified as a smoker. When ex-smokers are included in the non-smoking group, the model correctly classifies the two conditions in about 80% of cases. Conclusions: The results demonstrate that the models generated from pyrosequencing data are useful for identifying tobacco smokers and estimating an individual’s chronological age, particularly for younger subjects. Further studies are needed to develop models with higher predictive accuracy and to integrate these tools into regular forensic practice. Full article

Background/Objectives: Rapid DNA systems accelerate STR profiling but often require the consumption of the entire swab, limiting confirmation testing or downstream analyses. We previously validated a simple subsampling protocol for blood swabs on the RapidHIT^TM ID, using a rigid subungual mini-swab (Copan Italia S.p.A). A new version of this instrument has recently been released, featuring redesigned software and consumables. The RapidINTEL^TM Plus sample cartridge now enables two distinct lysis/extraction protocols, expanding analytical possibilities for rich biological traces. We evaluated subsampling performance using the subungual mini-swab and microFLOQ^® swabs (Copan Italia S.p.A), and assessed feasibility for both blood and buccal reference swabs. Methods: Whole blood from four donors was deposited onto regular Copan swabs (10 µL) or microFLOQ^® swabs (1 µL). A comparison was performed between the direct analysis of blood swabs using a RapidHIT^TM ID V1 (RapidINTEL^TM cartridge) and a RapidHIT^TM ID V2 (RapidINTEL^TM Plus cartridge, GENERAL protocol). Subsequently, both the GENERAL and SPECIALIZED protocols were tested after subsampling from primary blood or buccal swabs dried for 24 h using either a subungual mini-swab or a microFLOQ^®. Results: Blood-swab subsampling on the V2 produced usable STR profiles with both the subungual mini-swab and the microFLOQ^®. The subungual mini-swab was compatible with both the GENERAL and SPECIALIZED protocols. For blood applications, microFLOQ^® fiber treatment showed no inhibitory effects. Reference buccal swabs were successfully analyzed with the RapidINTEL^TM Plus cartridge, either directly (regular swab) or via subungual subsampling under both protocols. In contrast, in this feasibility dataset (single analysis per donor per condition), subsampling a reference swab with microFLOQ^® did not yield suitable profiles for RapidINTEL^TM Plus analysis under the tested conditions. Conclusions: This feasibility study indicates that the subsampling strategy can be applied on the RapidHIT^TM ID V2, particularly using subungual mini-swabs, to retain the primary swab for potential downstream testing while maintaining usable STR profile quality for blood and buccal reference workflows under the tested conditions. Full article

This review explores the vital role of orthodontic specialty and orthodontic records in forensic investigations, emphasizing their contribution to human identification. It synthesizes current literature on methodologies, advantages, limitations, and future perspectives, positioning orthodontics as a valuable adjunct within the forensic toolkit. Collaboration between orthodontists, forensic scientists, and AI technologies will further strengthen the reliability and speed of human identification, reinforcing justice and humanitarian efforts worldwide. Full article

Burnout syndrome is increasingly recognized as a significant occupational health issue, characterized by emotional exhaustion, depersonalization, and a reduced sense of personal accomplishment. It predominantly arises from chronic work-related stress, but recent research has highlighted the role of genetic and epigenetic factors in determining individual vulnerability to burnout. This review aims to synthesize findings regarding the genetic footprints of burnout, focusing on genes related to stress regulation, including the 5-HTT (serotonin transporter) gene, BDNF (brain-derived neurotrophic factor) gene, and NR3C1 (glucocorticoid receptor) gene. Twin studies reveal that burnout is moderately heritable, with genetic factors accounting for 33–36% of the variability in burnout-related traits, such as emotional exhaustion and performance-based self-esteem. However, burnout risk seems highly driven by non-shared environmental factors, such as work stress, lack of social support, and personal coping mechanisms. Specific genetic polymorphisms in the serotonergic system (5-HTT) and HPA axis genes (NR3C1, FKBP5) have been linked to increased burnout susceptibility, particularly in individuals exposed to chronic job strain or early-life stressful situations. Variations in 5-HTT rs6354 and HTR2A rs6313 are associated with altered stress reactivity, while polymorphisms in NR3C1 and FKBP5 contribute to dysregulation of the HPA axis, which influences cortisol secretion patterns in response to stress. Increased methylation in genes like BDNF and SLC6A4 has been observed in individuals with burnout, suggesting that environmental stressors may lead to lasting changes in gene expression, contributing to the syndrome’s development. Studies on telomere length have shown that burnout is associated with accelerated cellular aging, with individuals exhibiting shorter telomeres, particularly during high-stress periods. These findings hold particular relevance for professionals within the forensic and justice systems, including law enforcement, the judiciary, and forensic experts, who operate under chronic, high-stakes stress. We examine how understanding the biological basis of burnout can inform more objective ‘fitness-for-duty’ evaluations and provide a scientific framework for distinguishing physiological exhaustion from professional negligence in legal contexts. Full article

Background/Objectives: Hanging is the most common method of suicide in most countries worldwide. It is characterized by high lethality, technical simplicity, and typical autopsy findings. Autopsy plays a crucial role in determining the cause and mechanism of death. While external injuries are relatively consistent, internal findings show considerable variability in the literature. The aim of this prospective study was to analyze 660 cases of suicidal hanging over a ten-year period, focusing on the occurrence of forensically relevant internal autopsy findings. Methods: The study was conducted at the Department of Forensic Medicine in Bratislava between 2015 and 2024. All cases underwent standardized complete autopsy, including histology, toxicology, and analysis of death circumstances. Recently reported thoracic aortic adventitial hemorrhages described in 2024 were evaluated only in a targeted subset of cases examined between July and December 2024. Statistical evaluation was performed using the chi-square test to identify associations between internal findings and suspension type, knot location, age, sex, and body weight. Results: The argent line was present in 61.1% of cases, most frequently with posterior knot placement and complete suspension. Neck muscle hemorrhages occurred in 53.8%, predominantly at the periosteal-clavicular attachment of the sternocleidomastoid muscle, with higher incidence in complete and anterior suspension. Amussat’s sign was observed in 10.2% of cases, and Etienne-Martin’s sign in 1.1%. Fractures of the laryngo-hyoid complex were present in 49.7%, mainly in cases with complete suspension and posterior knot location. Cervical spine injuries were detected in 2.6%, predominantly in older males and with anterior knot placement. Simon’s hemorrhages occurred in 35.2%, mainly in younger individuals and complete suspension. Hemorrhages in the intestinal wall were detected in 7.4%, and rectal hemorrhages in 1.1% of cases. In the targeted 2024 subset, no thoracic aortic adventitial hemorrhages were identified. Conclusions: The findings suggest the forensic relevance of several internal findings associated with hanging, while emphasizing that the results were obtained using a uniform and consistently applied autopsy protocol. They also indicate the need for further research, particularly regarding recently reported adventitial hemorrhages of the thoracic aorta, which were assessed only in a limited subset of cases during the final months of the study and were not identified in our material. Full article

Background and Clinical Significance: Fatal attacks by domestic dogs, particularly against familiar owners, are rare but represent a significant forensic and public health concern. Understanding the dynamics and forensic features distinguishing predatory aggression from postmortem scavenging is essential, especially when cases involve large breeds and vulnerable individuals, such as the elderly. Case Presentation: An 82-year-old man was found dead in his home, presenting extensive mutilation of the head and neck. He lived with his daughter and her four-year-old male German Shepherd. The dog exhibited blood and tissue residues on its mouth, forelimbs, and abdomen. Autopsy findings indicated death due to hemorrhagic shock from deep cervical and facial vessel lacerations. There were no defense wounds or classic bite marks; however, massive excision of musculocutaneous tissue from the neck to the scalp suggested active predation rather than postmortem scavenging. The facial and cervical soft tissues were completely avulsed, exposing deep anatomical structures and causing the loss of the right eye and dental elements. No signs of third-party involvement, intrusion, or external aggression were identified, and the dog appeared healthy and unrestrained. Conclusions: The findings support the theory of a fatal attack by the household German Shepherd occurring shortly after the victim’s meal. This case underscores the potential for lethal aggression in domestic dogs toward familiar humans, even in the absence of provocation or prior behavioral concerns. It highlights the importance of awareness and preventive measures when managing large, powerful dog breeds in environments with elderly or otherwise vulnerable individuals. Full article

Background/Objectives: Nasal morphology is an important craniofacial feature in forensic anthropology for examining population-level variation and informing biological profiling in living individuals. Despite genetic, historical, and cultural similarities between Malaysian and Indonesian populations, population-specific nasal anthropometric data remain limited. This study integrates nasal anthropometry with multivariate analytical approaches to examine population- and sex-related variation in nasal morphology among Malaysian and Indonesian adults. Methods: A cross-sectional anthropometric study was conducted among Malaysian and Indonesian adults. Nasal breadth and nasal height were measured using standardised techniques, and the nasal index was calculated. Univariate analyses were performed to assess population- and sex-related differences. Multivariate analyses, including principal component analysis (PCA), hierarchical cluster analysis (HCA), and partial least squares (PLS), were applied to eleven nasal parameters and the nasal index to evaluate patterns of population differentiation. Results: Malaysian participants predominantly exhibited leptorrhine nasal types, whereas mesorrhine nasal types were more prevalent among Indonesian participants. Significant differences were observed in nasal height and nasal index between populations, while nasal breadth did not differ significantly. Sex-related variation was evident, with males generally exhibiting larger nasal dimensions and higher nasal indices than females. Multivariate analyses demonstrated clear population separation, with modelling indicating that population group contributed more strongly to morphological differentiation than sex. Conclusions: The findings provide population-specific soft-tissue nasal data for Malaysian and Indonesian groups and support the use of multivariate analytical frameworks in forensic anthropological research. Full article

The application of advanced imaging techniques, particularly computed tomography (CT), photogrammetric scanning, and three-dimensional reconstructions of body surfaces and skeletal remains, is becoming a crucial component of Forensic Anthropology. These tools enable a non-invasive and highly standardized analysis of both intact cadavers and human remains recovered from terrestrial or aquatic environments, providing reliable support in identification processes, traumatological reconstruction, and the assessment of taphonomic processes. In the context of estimating the Post-Mortem Interval (PMI) and the Post-Mortem Submersion Interval (PMSI), digital imaging allows for the objective and reproducible documentation of morphological changes associated with decomposition, saponification, skeletonization, and taphonomic patterns specific to the recovery environment. Specifically, CT enables the precise assessment of gas accumulation, transformations in residual soft tissues, and structural bone modifications, while photogrammetry and 3D reconstructions facilitate the longitudinal monitoring of transformative processes in both terrestrial and underwater contexts. These observations enhance the reliability of PMI/PMSI estimates through integrated models that combine morphometric, taphonomic, and environmental data. Beyond PMI/PMSI estimation, imaging techniques play a central role in anthropological bioprofiling, facilitating the estimation of age, sex, and stature, the analysis of dental characteristics, and the evaluation of antemortem or perimortem trauma, including damage caused by terrestrial or fauna. Three-dimensional documentation also provides a permanent, shareable archive suitable for comparative analyses, ensuring transparency and reproducibility in investigations. Although not a complete substitute for traditional autopsy or anthropological examination, imaging serves as an essential complement, particularly in cases where the integrity of remains must be preserved or where environmental conditions hinder the direct handling of osteological material. Future directions include the development of AI-based predictive models for PMI/PMSI estimation using automated analysis of post-mortem changes, greater standardization of imaging protocols for aquatic remains, and the use of digital sensors and multimodal techniques to characterize microstructural alterations not detectable by the naked eye. The integration of high-resolution imaging and advanced analytical algorithms promises to further enhance the reconstructive accuracy and interpretative capacity of Forensic Anthropology. Full article

Emerging contaminants (ECs) are trace-level chemical and biological compounds detected in the environment, particularly in water, including personal care products, microplastics, nanoplastics, antibiotic resistance genes, etc., which have the potential to endanger ecosystems and human health. Forensic techniques, which are used to detect and trace the sources of ECs with specificity and accuracy, like gas and liquid chromatography coupled with mass spectrometry, Fourier transform infrared spectroscopy, inductively coupled plasma mass spectrometry, isotope ratio mass spectrometry, remote sensing, geographical information systems, and statistical and machine learning approaches are discussed in this study. The present analysis also outlines the types of emerging contaminants, along with their possible sources, including domestic and municipal wastewater, wastewater from healthcare institutions, urban runoff and stormwater, industrial and commercial discharges, effluents from agricultural and animal husbandry, and pollution from recreational activities in water. The review concludes by emphasizing the future need for real-time detection systems, integrated data analysis, and stronger regulatory frameworks to manage ECs more effectively. Full article

Background: In forensic ballistics, identifying ammunition types on physical evidence is critical, particularly when metallic residues are minimal. This study comparatively analyzes the elemental signatures deposited by two common projectiles—Full Metal Jacket (FMJ) (Cu/Zn jacket) and Lead Round Nose (LRN) (exposed Pb core)—on complex targets, including pig bone/tissue and mango wood. Methods: Using a semi-automatic handgun at an intermediate range of 5.0 m, residues were examined through high-resolution benchtop Micro-XRF (M4 Tornado) for micro-spatial analysis and Portable XRF (Elio) for rapid field characterization. Additionally, fresh pork leg samples were subjected to a 3-month environmental degradation period to assess trace persistence. Results: Observations indicated that LRN projectiles exhibit markedly elevated Lead (Pb) concentrations along the wound track in bone, hence confirming Pb as a reliable indicator for unjacketed ammunition; specifically, the median Pb concentrations at bullet wiping were 10.39 wt% for M4 and 7.34 wt% for Elio. Conversely, FMJ traces remain strictly confined to the surface bullet wipe area, with median concentrations of Pb, Cu, and Zn being 2.21 wt%, 0.24 wt%, and 0.59 wt% via M4, respectively. Statistical analysis showed a strong correlation for high-concentration elements on tissue, but significantly greater variance on wooden surfaces where FMJ traces exhibited a very weak negative correlation (r = −0.2774) due to minimal and irregular metal transfer. Taphonomic evaluation revealed that the Pb signature from LRN is exceptionally stable (r ≈ 0.9999) even after decomposition, while FMJ signatures are highly sensitive to environmental exposure. Conclusions: This research underscores the necessity of high-sensitivity Micro-XRF (M4) for definitive ammunition verification, providing a refined analytical framework for shooting incident reconstruction even involving degraded remains or complex environmental scenes. Full article

Background/Objectives: The identification of individuals from human remains is crucial in any scenario where their identity is unknown. The study of ontogenetic allometry, which refers to proportional changes in the shape and size of bones during growth, provides important baseline information for constructing biological profiles. Methods: This study focuses on the analysis of the ontogenetic allometry of the scapula in Portuguese reference skeletal samples, using geometric morphometric techniques. The sample includes 140 individuals (67 females, 73 males), ranging from birth to 89 years old. Scapulae were photographed, and seven landmarks and forty semi-landmarks were digitized using the “tps” programs. Statistical analyses were performed using the MorphoJ (v. 1.08.02) and PAST (v. 5.2) programs. Results: The results point to a significant and continuous growth of the scapula in the early stages of life, with a tendency to stabilize after adolescence. Centroid size significantly influenced shape variation across the full sample. Conclusions: These findings provide a descriptive baseline of scapular development that can aid future anthropological and forensic research, including studies on population variation and age-related morphological trajectories. Full article

Background/Objectives: MDMA and MDA are among the stimulant drugs most frequently encountered in forensic casework, and oral fluid represents a practical biological matrix for their detection. However, liquid oral fluid requires refrigeration, is susceptible to degradation, and can be logistically demanding for routine laboratories. Dried Oral Fluid Spots (DOFS) offer a low-cost and stable alternative. This study aimed to develop and validate a DOFS-based analytical workflow for quantifying MDMA and MDA using liquid chromatography and a diode-array detector. Methods: Watercolor paper was selected as the substrate and pretreated with diluted nitric acid to improve analyte desorption. DOFS were prepared using 150 µL of pooled oral fluid, dried for 4 h, and extracted with methanol. Chromatographic separation was performed on a phenyl column using aqueous TFA and acetonitrile mobile phase. Method validation followed the ICH M10 criteria. Results: The method showed linear responses between 12.5 and 5000 ng mL⁻¹, with LOD and LLOQ of 6 and 12 ng mL⁻¹ for both analytes, respectively. Precision and accuracy met acceptance criteria across all QC levels. Recoveries ranged from 84% to98%. DOFS samples demonstrated adequate stability under multiple storage and handling conditions. Conclusions: The optimized DOFS–LC–DAD workflow offers a robust, low-cost, and flexible approach for the analysis of MDMA and MDA in oral fluid for laboratory-based or semi-controlled collection environments. Its compatibility with both LC- and GC-based detectors enhances applicability in diverse forensic laboratory settings. Full article

Forensic anthropology, a specialized branch of biological anthropology, plays a crucial role in the identification of human remains, particularly when conventional methods such as fingerprinting are not applicable. In Ecuador, its relevance has increased in response to challenges such as intentional deaths, forced disappearances, violence, mass fatalities, and migration-related deaths. Despite its growing importance, the field faces significant limitations, including restricted access to advanced technologies, limited training opportunities for local forensic anthropologists, and insufficient resources for research and the application of advanced methodologies for victim identification. This article examines the development and current state of forensic anthropology in Ecuador, emphasizing the urgent need for population-specific standards, the establishment of a national osteological collection, and stronger institutional support. It also highlights the contributions of bioarchaeological research and its potential to enhance forensic practices. By analyzing the challenges of identifying skeletonized human remains and other instances of human rights violations, the study underscores the necessity of advancing forensic anthropology in the country. The article further discusses how interdisciplinary efforts have contributed to forensic knowledge in Ecuador and concludes by emphasizing the importance of ethical guidelines, technological integration, and improved infrastructure to strengthen forensic anthropology as both a scientific discipline and a humanitarian tool. Full article

Background: Cigarette burns represent a well-established forensic indicator of inflicted injury, frequently encountered in cases of domestic violence. Clinical significance: Their morphological consistency and anatomical distribution offer valuable elements for differentiating between intentional and accidental trauma. Case Presentation: In this study, we report the case of a 40-year-old woman who presented with multiple cutaneous lesions attributed to repeated assaults by her intimate partner. The forensic medical examination revealed five discrete scars characterized by sharply demarcated borders, circular to oval shapes, and dimensions ranging from 0.7 to 1.5 cm. These lesions were anatomically located in regions not typically accessible for self-infliction. To reinforce the diagnostic interpretation and assess reproducibility, a controlled experimental protocol was conducted using porcine skin matrices. Cigarette burns were recreated under variable conditions of contact pressure and exposure duration. The lesions produced on the biological substrate exhibited morphological features consistent with those observed in the patient, suggesting compatibility with cigarette-induced thermal injury. Conclusions: These findings provide circumstantial support for the forensic interpretation but must be considered within the limitations of the experimental model. This integrated approach underscores the relevance of combining clinical forensic documentation with experimental validation to support medico-legal conclusions in cases of suspected interpersonal violence. Full article

Background/Objectives: This systematic review examines how artificial intelligence (AI) is transforming fingerprint and latent print identification in criminal investigations, tracing the evolution from traditional dactyloscopy to Automated Fingerprint Identification Systems (AFISs) and AI-enhanced biometric pipelines. Methods: Following PRISMA 2020 guidelines, we conducted a literature search in the Scopus, Web of Science, PubMed/MEDLINE, and legal databases for the period 2000–2025, using multi-step Boolean search strings targeting AI-based fingerprint identification; 68,195 records were identified, of which 61 peer-reviewed studies met predefined inclusion criteria and were included in the qualitative synthesis (no meta-analysis). Results: Across the included studies, AI-enhanced AFIS solutions frequently demonstrated improvements in speed and scalability and, in several controlled benchmarks, improved matching performance on low-quality or partial fingerprints, although the results varied depending on datasets, evaluation protocols, and operational contexts. They also showed a potential to reduce certain forms of examiner-related contextual bias, while remaining susceptible to dataset- and model-induced biases. Conclusions: The evidence indicates that hybrid human–AI workflows—where expert examiners retain decision making authority but use AI for candidate filtering, image enhancement, and data structuring—currently offer the most reliable model, and emerging developments such as multimodal biometric fusion, edge computing, and quantum machine learning may contribute to making AI-based fingerprint identification an increasingly important component of law enforcement practice, provided that robust regulation, continuous validation, and transparent governance are ensured. Full article

Background. Determining the cause and manner of death in scenes involving multiple and putrified bodies found in the same environment is a real challenge for forensic pathologists. While common scenarios include fires, vehicle crashes, and natural disasters, one of the most common causes is drug intoxication or poisoning, and the scene must be carefully evaluated based on circumstantial evidence. Carbon monoxide (CO) (also called “the silent killer”) remains one of the leading agents capable of producing simultaneous fatalities. In multi-body scenes, distinguishing between homicide–suicide, double suicide, and accidental deaths adds further complexity. The aim of this study is to highlight the limitations of toxicological and pathological investigations in advanced putrefaction and to emphasize the role of scene investigation in the interpretation of suspected CO-related deaths. Methods. The authors report a case of suspected CO intoxication involving two bodies in an advanced stage of decomposition recovered from the same room. The scene investigation, coupled with the presence of a malfunctioning combustion source, raised suspicion of CO exposure; however, analytical interpretation was severely constrained by the altered condition of biological samples. Results. Advanced decomposition magnifies these challenges. Putrefactive changes can mimic traumatic injuries, hide hypostasis, and compromise both macroscopic and microscopic evaluations due to autolysis and gas formation. Toxicological investigations are frequently hindered by the degradation or absence of key biological matrices such as blood, cavity fluids, or vitreous humor, rendering carboxyhaemoglobin quantification unreliable or impossible. These limitations may lead to incorrect medico-legal conclusions. Conclusions. Determining the cause and manner of death in complex multi-body scenes requires careful evaluation of circumstantial evidence and scene investigation, particularly when advanced decomposition compromises biological analyses and toxicological interpretation. Full article

Background/Objectives: A common challenge in both forensic and bioarchaeological research is commingling, the intermixing of skeletal material originating from multiple individuals or contexts. To tackle that problem past reassociation methods primarily relied on visual assessment or metric comparisons. However, recent advances in geometric morphometrics show strong potential for improving the sorting of commingled remains. This study applies a three-dimensional (3D) geometric morphometric method to evaluate its effectiveness in reassociating adjoining thoracic vertebrae. Methods: Two vertebral pairs, T4–T5 and T5–T6, from 65 and 73 individuals, respectively, were analyzed. These pairs were chosen due to limited anatomical variability, while they were also the most consistently preserved pairs. All specimens were scanned using a structured-light 3D scanner, and the dataset was derived from three Greek skeletal collections representing different geo-chronological contexts. Fourteen anatomical landmarks were placed on the superior rim and articular facets of the lower vertebra and mirrored onto the lower rim and facets of the adjoining upper vertebra. To remove the size effects the landmark coordinates were converted to Procrustes coordinates, while examining morphological similarity was quantified using Euclidean distances. For each pair, the vertebrae with the smallest Euclidean distances were considered the most probable true anatomical matches. Results: The correct T4–T5 match fell within the three smallest distances in 66.2% of cases, while for the T5–T6 pair, correct matches were found between the first three possible matches in a percentage of 43.8%. These findings indicate that the method can eliminate roughly 50–70% of incorrect matches and therefore narrow the plausible pairings. Conclusions: Future research incorporating more pairs and an expanded landmark dataset may result in greater accuracy for reassociation with 3D geometric morphometrics. Full article

Background: Postmortem DNA identification of highly decomposed human remains is often limited by the availability and quality of conventional biological samples. Keratinized tissues, such as fingernails, represent a potentially valuable alternative due to their anatomical resistance to environmental degradation, however, their use as primary biological material for DNA profiling remains underreported in forensic practice. Case presentation: We report a case involving the recovery of a highly decomposed body of a missing woman, in which DNA samples were collected from a fingernail and a tooth. DNA extraction was performed using the PrepFiler Forensic DNA Extraction Kit for the fingernail sample and PrepFiler BTA Forensic DNA Extraction Kit for the tooth sample. No usable DNA profile was obtained from the tooth sample; however, the fingernail sample yielded a complete and high-quality STR profile with successful amplification across all 24 loci (GlobalFiler PCR Amplification Kit). Reference buccal swabs from the presumed biological parents were collected for subsequent kinship analysis. Discussion: Kinship analysis based on allele frequencies in the Croatian population resulted in a combined paternity index (CPI) corresponding to a probability of paternity of 99.99999812%, providing strong genetic support for the proposed identity of the deceased. Notably, this is the first documented forensic case in Croatia in which nail material served as the primary—and ultimately successful—biological sample for postmortem identification. Conclusions: This case highlights the evidentiary value of fingernails as a robust, accessible, and forensically valid DNA source in postmortem identification, particularly in cases of advanced decomposition where conventional biological materials are unavailable or degraded. Further studies involving larger sample sets and diverse postmortem conditions are needed to support the broader implementation of nail material in routine forensic identification workflows, particularly within the Croatian medico-legal context. Full article

Background/Objectives: Snakebite envenoming remains a critical yet frequently under-recognized cause of mortality in many parts of the world, particularly in tropical and rural areas where access to timely medical care and accurate post-mortem investigation is limited. While clinical and epidemiological data on snakebites have been extensively studied, the forensic characterization of fatal envenomations remains fragmentary and inconsistently documented. This review aims to synthesize the existing literature on autopsy-confirmed snakebite deaths, focusing on the pathological and toxicological evidence that supports cause-of-death determinations in forensic settings. Methods: A comprehensive search of the PubMed NCBI databases identified nine relevant studies, including case reports, retrospective analyses, and systematic reviews. Results: Across these reports, a range of lethal mechanisms were identified, including venom-induced consumption coagulopathy (VICC), acute renal failure (frequently in the setting of rhabdomyolysis and acute tubular necrosis), neurotoxic respiratory arrest, multi-organ necrosis, and myocardial infarction. Histological findings frequently revealed glomerular and tubular necrosis, pulmonary edema and/or hemorrhage, pituitary and adrenal hemorrhage, and cerebral ischemic changes. Toxicological confirmation was achieved in several cases using ELISA and liquid chromatography–mass spectrometry (LC–MS/MS), underscoring the importance of biochemical validation in post-mortem diagnosis and the value of analytical tools beyond ELISA (e.g., immunoaffinity LC–MS/MS, venom-specific immunohistochemistry, zymography for SVMP activity). Conclusions: Our findings highlight the variability in venom effects across snake families—particularly Viperidae, Elapidae, and Lamprophiidae/Atractaspididae—and emphasize the indispensable role of forensic autopsy in distinguishing snakebite envenoming from other causes of sudden or unexplained death. However, significant limitations persist, including inconsistent autopsy protocols, lack of species-specific venom assays, and poor integration of toxicological methods in routine forensic practice. Addressing these gaps through standardized forensic guidelines and improved access to diagnostic tools is essential for enhancing the accuracy of death investigations in envenoming-endemic regions. Full article