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Forensic microbiology is a relatively new discipline, born in part thanks to the development of advanced methodologies for the detection, identification and characterization of microorganisms, and also in relation to the growing impact of infectious diseases of iatrogenic origin. Indeed, the increased application of medical practices, such as transplants, which require immunosuppressive treatments, and the growing demand for prosthetic installations, associated with an increasing threat of antimicrobial resistance, have led to a rise in the number of infections of iatrogenic origin, which entails important medico-legal issues. On the other hand, the possibility of detecting minimal amounts of microorganisms, even in the form of residual traces (e.g., their nucleic acids), and of obtaining gene and genomic sequences at contained costs, has made it possible to ask new questions of whether cases of death or illness might have a microbiological origin, with the possibility of also tracing the origin of the microorganisms involved and reconstructing the chain of contagion. In addition to the more obvious applications, such as those mentioned above related to the origin of iatrogenic infections, or to possible cases of infections not properly diagnosed and treated, a less obvious application of forensic microbiology concerns its use in cases of violence or violent death, where the characterization of the microorganisms can contribute to the reconstruction of the case. Finally, paleomicrobiology, e.g., the reconstruction and characterization of microorganisms in historical or even archaeological remnants, can be considered as a sister discipline of forensic microbiology. In this article, we will review these different aspects and applications of forensic microbiology.

Background: In forensic pathology, asphyxia presents a problem as frequently as it is thorny and challenging. Some knowledge in forensic pathology is still considered to be traditionally acquired and is not critically assessed with modern statistical or technical tools. In this study, we seek to examine the injuries that are considered to be typical of cases of hanging (neck lesions, haemorrhages, and pleural and epicardial petechiae). Methods: We evaluated whether there was, indeed, a statistically significant association between these injuries and deaths from hanging as compared to non-hanging deaths. We collected 399 cases (32 deaths by hanging and 367 cases of non-hangings), built contingency tables and performed chi-square tests for each variable (lesion) examined; we also analysed this association in various subgroups of the sample (according to sex, age and weight ranges). Results: Our results did not deviate from the expected outcome based on traditional knowledge, although they do provide a more detailed demonstration and clarification of traditional knowledge regarding this topic. Conclusions: These findings provide valuable insights for future discussion, examination and deepening of knowledge that is traditionally accepted but often undersupported in the literature.

In the global migration crisis, one of the challenges in the effort to identify deceased migrants is establishing their region of origin, which facilitates the search for ante mortem data to be compared with the post mortem information. This pilot study explores the potential of using stable isotope analysis to distinguish between individuals coming from West Africa and the Horn of Africa. Six individuals (four of known origin and two of unknown origin) were sampled. δ13CVPDB(keratin), δ15NVPDB(keratin) and δ18OVSMOW(keratin) of hair were analysed using Elemental Analyzers coupled with Isotope Ratio Mass Spectrometry (IRMS). δ18OVSMOW(carbonate) and δ13CVPDB(carbonate) of bone were analysed using GasBench II with IRMS, while 87Sr/86Sr composition was determined in bone and dental enamel using laser ablation multi-collector inductively coupled plasma mass spectrometry. The stable isotope compositions of the individual from the Horn of Africa differed from the other individuals. The differences found between 87Sr/86Sr of enamel and bone and between δ18O and δ13C in bone and hair reflect changes in sources of food and water in accordance with regionally typical migration journeys. The analysis of multiple stable isotopes delivered promising results, allowing us to narrow down the region of origin of deceased migrants and corroborate the information about the migration journey.

In Italy, in only 2018, 3310 people died in road traffic accidents, more than in any other European country. Since the revelation of this occurrence, the authors carried out an analysis aimed at investigating if there was a difference in the injury patterns among different road users. A retrospective post-mortem study on road traffic fatalities was performed, which had been autopsied at the Institute of Forensic Medicine of Milan. First, the authors analyzed the epidemiological data of all the 1022 road traffic accidents subjected to an autopsy from 2007 to 2019. Secondly, further analysis of individual autopsy reports was carried out. For this purpose, 180 autopsies belonging to 5 different categories were analyzed: car, pedestrian, motorbike, bicycle, and truck. Seventy-six percent of road traffic fatalities were male, 54% were between 10 and 49 years of age, and 62% of the patients died before arriving at a hospital. “Multiple injuries” was the main cause of death. Traumatic brain injuries were particularly high in pedestrians and cyclists. In car, motorbike, and truck fatalities, thoracic and abdominal injuries were the most frequent. Therefore, pedestrians and cyclists had a higher prevalence for traumatic head injuries, while car, motorcycle, and truck occupants, on the other hand, had a higher prevalence for thoracic and abdominal injuries.

Paranasal sinuses represent one of the most individualizing structures of the human body and some of them have been already analyzed for possible applications to personal identification, such as the frontal and sphenoid sinuses. This study explores the application of 3D–3D superimposition to maxillary sinuses in personal identification. One hundred head CT-scans of adult subjects (equally divided among males and females) were extracted from a hospital database. Maxillary sinuses were segmented twice from each subject through ITK-SNAP software and the correspondent 3D models were automatically superimposed to obtain 100 matches (when they belonged to the same person) and 100 mismatches (when they were extracted from different individuals), both from the right and left side. Average RMS (root mean square) point-to-point distance was then calculated for all the superimpositions; differences according to sex, side, and group (matches and mismatches) were assessed through three-way ANOVA test (p < 0.017). On average, RMS values were lower in matches (0.26 ± 0.19 mm in males, 0.24 ± 0.18 mm in females) than in mismatches (2.44 ± 0.87 mm in males, 2.20 ± 0.73 mm in females) with a significant difference (p < 0.001). No significant differences were found according to sex or side (p > 0.017). The study verified the potential of maxillary sinuses as reliable anatomical structures for personal identification in the forensic context.

Age estimation in juveniles is a critical procedure in judicial cases for verification of imputability or for civil reasons when adopting children. Several methods based both on skeletal and dental growth have been performed and applied on different populations; nevertheless, few articles have compared different methods in order to test their reliability in different conditions and age ranges, and this is a clear obstacle in the creation of common guidelines for age estimation in the living. A comparison of five dental methods (Anderson, Ubelaker, Schour and Massler, Gustafson and Koch, Demirjian) and one skeletal method (Greulich a Pyle atlas) was performed on a population of 94 children aged between 0 and 8 years. Results showed that, whereas under 2 years all the methods have the same inaccuracy, over 2 years the diagram methods, such as Schour and Massler and Ubelaker’s revised one, have a lower error range than the most frequently used Greulich and Pyle atlas and Demirjian method. Schour and Massler, Gustafson and Koch, and Ubelaker methods showed, respectively, a mean error amounting to 0.40, 0.53, and 0.56 years versus the 0.74 and 0.88 years given by Demirjian and the Greulich and Pyle atlas. An in-depth analysis of the potential of several methods is necessary in order to reach a higher adherence of age estimation with the complexity of growth dynamics.

Structural violence is an indirect form of violence that can lead to physiological consequences. Interestingly, these physiological disruptions may affect the skeletons and can therefore provide relevant information on violence and way of life in the analysis of skeletal remains. The aim of the present study was to test the hypothesis that migrants who died in the Mediterranean Sea would present physiological cranial stress markers such as cribra orbitalia (CO), porotic hyperostosis (PH), and linear enamel hypoplasia (LEH) more frequently and more severely than Italians of the 20th century. With this intent, a total of 164 crania were examined: 139 from deceased migrants recovered from a shipwreck in the Mediterranean Sea in 2015, aged between 16 and 35 years old, and 25 of the same age from the CAL Milano Cemetery Skeletal Collection. Both presence and severity of CO, PH, and LEH were evaluated. The data obtained were analyzed using Wilcoxon signed-rank and independence Chi-squared tests to compare the results between the two samples and to test whether there was an association between the sample of migrants and the occurrence of lesions. As a result, CO and PH appeared more frequently and more severely in the migrant sample. In addition, migrants were significantly associated with CO, PH, and LEH (p-values < 0.05). Although this does not imply in any way that CO, PH, and LEH are specific to migration, they should be regarded as indicators of structural violence.

The permanence of a buried body in soil always induces the formation of a decomposition island that can serve as a significant recording location to understand how the persistence of a clandestine grave affects soil. This study aims to analyze the elemental exchange from buried bodies to soil, with a focus on phosphorus content, and to determine the effects of environmental factors on its persistency. The experiment was carried out using eleven swine carcasses buried in an open site (northern Italy). The analyses were performed using the Olsen P method, which allowed for a recognition of the trend of the amount of phosphorus over time, due to the decomposition of phospholipids, followed by the transfer of the element from bone to soil. Additionally, microanalyses performed using a scanning electron microscope (SEM-EDS) on two different soil sample specimens (i.e., “dust” and “plug”) allowed for the identification of numerous phosphatic features (i.e., coatings, infillings, impregnations, and organo-mineral associations), which are the result of the interaction between soil and body fluids and can thus be used as indicators of the former presence of decomposing body (even in its absence). The ultramicroscopic analysis also shows increasing and decreasing amounts of P2O5 over time in the soil, which could be related to environmental conditions (i.e., soil moisture), due to the leaching of phosphorus induced by the percolation of natural rainwater. The study underlines the potential use of these methods to evaluate the possibility of a cadaver–soil linkage and of assessing the burial in the soil for a variable period. Moreover, the study may aid in analyzing the dynamics of phosphorus migration from buried bodies to soil during and after the decomposition process.

Virtual anthropology (VA) has recently produced an additional tool for the analysis of commingled remains and is based on the distance analysis between three-dimensional (3D) models of bones. To date, the pair-matching of the innominate bone through a 3D approach remains partially unexplored. Here, 44 abdominal CT scans (22 males and 22 females) were selected from a hospital database, and the pubic bones were segmented through ITK-SNAP software. The models were hollowed with Viewbox4 to minimize the amount of trabecular bone. The left pubic bones were mirrored and superimposed on the right ones, according to the smallest point-to-point difference between the two surfaces through VAM software. RMS distances between models were calculated through VAM, producing RMS values for 20 matches and 420 mismatches for each sex group. Differences in RMS distance values between matches and mismatches were investigated through Mann–Whitney tests (p < 0.05); the repeatability of the procedure was assessed through absolute and relative technical error measurement (TEM and rTEM). RMS distance values of matches and mismatches were significantly different (p < 0.01) in both groups. The method yielded optimal results with high sensitivity (100.0%) and specificity (99.8% in males, 98.8% in females) rates according to the chosen threshold. This project contributes to the research field of VA with a valuable adjunct that may bolster and strengthen the results of the current visual and osteometric methods through a multidisciplinary approach.

During the COVID-19 pandemic, not only crowded refugee camps and immigration detention centers, but also receptions were places in which outbreaks occurred. To date there has been no report of the application of a COVID-19 surveillance system in reception centers for unaccompanied foreign minors only, who most of all deserve the utmost attention. Aware of this critical issue, we implemented a pilot COVID-19 surveillance program at the Zendrini center in Milan. It was started in September 2021 and was carried out for 4 months. Nasopharyngeal antigenic swabs were adopted. One day a week, two forensic physicians performed the first antigenic swab to minors who had just entered the center, or a monitoring swab after 15 days to those who were still hosted at the center. Operators were also swabbed for surveillance. A total of 80 subjects were enrolled and divided into 68 (72.5%) unaccompanied foreign minors and 22 (27.5%) operators. A total of 178 antigenic nasopharyngeal swabs were performed and tested negative. Regarding the monitoring activities, it was found that the minimum number of swabs per subject was 1 and the maximum number was 7, with an average value of 2.2 per individual. Having been able to confirm the absence of SARS-CoV-2 within the community represented a way to protect individual and collective health that could not have been pursued otherwise. Only inclusive approaches can allow communities and societies to respond more effectively to this crisis, and reduce the risk of future ones, intended as both upcoming COVID-19 waves and new infectious diseases.

Although not without subjectivity, the cranial trait scoring method is an easy visual method routinely used by forensic anthropologists in sex estimation. The revision presented by Walker in 2008 has introduced predictive models with good accuracies in the original populations. However, such models may lead to unsatisfactory performances when applied to populations that are different from the original. Therefore, this study aimed to test the sex predictive equations reported by Walker on a contemporary Italian population (177 individuals) in order to evaluate the reliability of the method and to identify potential sexual dimorphic differences between American and Italian individuals. In order to provide new reference data to be used by forensic experts dealing with human remains of modern/contemporary individuals from this geographical area, we designed logistic regression models specific to our population, whose accuracy was evaluated on a validation sample from the same population. In particular, we fitted logistic regression models for all possible combinations of the five cranial morphological traits (i.e., nuchal crest, mastoid process, orbital margin, glabella, and mental eminence). This approach provided a comprehensive set of population-specific equations that can be used in forensic contexts where crania might be retrieved with severe taphonomic damages, thus limiting the application of the method only to a few morphological features. The results proved once again that the effects of secular changes and biogeographic ancestry on sexual dimorphism of cranial morphological traits are remarkable, as highlighted by the low accuracy (from 56% to 78%) of the six Walker’s equations when applied to our female sample. Among our fitted models, the one including the glabella and mastoid process was the most accurate since these features are more sexually dimorphic in our population. Finally, our models proved to have high predictive performances in both training and validation samples, with accuracy percentages up to 91.7% for Italian females, which represents a significant success in minimizing the potential misclassifications in real forensic scenarios.

Forensic anthropologists rely on a number of parameters when analyzing human skeletal remains to assist in the identification of the deceased, predominantly age-at-death, sex, stature, ancestry or population affinity, and any unique identifying features. During the examination of human remains, it is important to be aware that the skeletal features considered when applying anthropological methods may be influenced and modified by a number of factors, and particular to this article, prescription drugs (including medical and non-medical use) and other commonly used drugs. In view of this, this paper aims to review the medical, clinical and pharmacological literature to enable an assessment of those drug groups that as side effects have the potential to have an adverse effect on the skeleton, and explore whether or not they can influence the estimation of age-at-death, sex and other indicators of the biological profile. Moreover, it may be that the observation of certain alterations or inconsistencies in the skeleton may relate to the use of drugs or medication, and this in turn may help narrow down the list of missing persons to which a set of human remains could belong. The information gathered from the clinical and medical literature has been extracted with a forensic anthropological perspective and provides an awareness on how several drugs, such as opioids, cocaine, corticosteroids, non-steroidal anti-inflammatory drugs, alcohol, tobacco and others have notable effects on bone. Through different mechanisms, drugs can alter bone mineral density, causing osteopenia, osteoporosis, increase the risk of fractures, osteonecrosis, and oral changes. Not much has been written on the influence of drugs on the skeleton from the forensic anthropological practitioner perspective; and this review, in spite of its limitations and the requirement of further research, aims to investigate the current knowledge of the possible effects of both prescription and recreational drugs on bones, contributing to providing a better awareness in forensic anthropological practice and assisting in the identification process of the deceased.

Insect-related evidence must be considered of probative value just as bloodstains, fingerprints, fibers, or any other materials. Such evidence if properly collected and analyzed can also provide useful details in the reopening of old unsolved murders, also called “cold cases”. This paper presents the case of two murders that occurred in two different European countries and remained unsolved for years. The remains of a girl found in Italy 17 years after her disappearance helped to solve a murder that occurred in Britain 8 years prior. The cases were unexpectedly linked together because of the similarities in the ritualistic placing of strands of hair and connections with the suspect. The trace evidence relating to insects and hairs played a relevant role in the conviction of the perpetrator. In Italy, the defense raised the doubt that the strands of hair found nearby the skeletal remains could be the result of insect feeding activity and not the result of a cut by sharp objects. Therefore, it was fundamental to distinguish between sharp force lesions and insect feeding activity on hair. This unusual application of insect-related evidence clearly emphasizes the importance of an appropriate professional collection and analysis of any physical evidence that could be of robust probative value.

Establishing the post mortem interval (PMI) is a key component of every medicolegal death investigation. Several methods based on different approaches have been suggested to perform this estimation. Among them, two methods based their evaluation on the effect of the temperature and time on the considered parameters: total body score (TBS)/accumulated degree-days (ADDs) and insect development. In this work, the two methods were compared using the results of minPMI and PMI estimates of 30 forensic cases occurring in northern Italy. Species in the family Calliphoridae (Lucilia sericata, Calliphora vomitoria and Chrysomya albiceps) were considered in the analyses. The results highlighted the limits of the TBS/ADD method and the importance of the entomological approach, keeping in mind that the minPMI is evaluated. Due to the fact that the majority of the cases occurred in indoor conditions, further research must also be conducted on the different taxa to verify the possibility of increasing the accuracy of the minPIM estimation based on the entomological approach.