Transcending time: the forensic anthropological case study of three unidentified transgender women in Italy in the early 1990s.

In forensic anthropology, the estimation of skeletal sex results in a dichotomous outcome: male or female. This poses challenges when dealing with transgender individuals. Italy has the highest rate per capita of transgender murders, making their identification a compelling issue. This study presents three cases of unidentified skeletal remains of transgender women (MtF) that underwent autopsies in the 1990s at the Institute of Legal Medicine of Milan. The transgender status of the individuals was determined from the autopsy records which indicated the co-existence of breast implants and male genitalia. The biological profile was created by estimating sex, age, ancestry, and stature, and the bones were investigated looking for evidence of Facial Feminization Surgery (FFS). Anthropological estimations revealed that the skeletons belong to male, adults, European individuals, between 164 and 184 cm. Fragmented gelatinous masses were found in association with two skeletons, which were interpreted as remnants of breast implants. In two cases, signs of remodeling of the cortical surface were observed on the zygomatic bones, although the bone marks observed here were not specific enough to link them to FFS. Despite some limitations, this study highlights the need for greater awareness among practitioners about this limitedly addressed issue, advocating for a more inclusive forensic anthropology that strives to improve methods and interpretation of evidence for the identification of transgender individuals.

Investigating gunshot wounds in charred bone with XRF spectroscopy: a technical note.

The analysis of traces of injuries can be difficult in cases of charred human remains since the alteration and fragmentation are high. The aim of this study is to explore the use of X-Ray Fluorescence (XRF) technique as a screening tool for detecting and analyzing gunshot residues (GSR) on cremated and highly fragmented materials, as it is a technique that allows for fast qualitative investigations without altering the sample or requiring sample preparation. The study was carried out on two steps: firstly, on completed skeletonized bones to verify if GSR survive to burning; secondly, we considered a more realistic situation, in which soft tissues were present before the shooting. To this aim, nine adult bovine ribs, four retaining soft tissue, five completely skeletonized, were subjected to a shooting test using two types of 9 mm projectiles (jacketed and unjacketed bullets). The ribs were then burnt until complete calcination in an electric furnace. The entry wound of each rib was analyzed using XRF, revealing traces of GSR. The XRF analysis showed that all samples, except for one, contain Pb and/or Sb near the lesion. Furthermore, the samples hit by unjacketed bullets had a more significant presence of Pb in macroscopic yellow areas, which persisted when moving away from the gunshot. These findings could pave the way for the use of XRF technology, mostly when a fast and immediate scan must be done on osteologic materials by a conservative method.

Skin surface debris as an archive of environmental traces: an investigation through the naked eye, episcopic microscope, ED-XRF, and SEM-EDX.

Traces from bodies can be of various nature, for example of biological or inorganic origin. Some of these historically have received more consideration than others in forensic practice. Samplings of gunshot residues or biological fluid traces are commonly standardized, whereas macroscopically invisible environmental traces are usually ignored. This paper simulated the interaction between a cadaver and a crime scene by placing skin samples on the ground of five different workplaces and inside the trunk of a car. Traces on samples were then investigated through different approaches: the naked eye, episcopic microscope, Scanning Electron Microscopy (SEM) with Energy-dispersive X-ray spectroscopy (EDX) and Energy Dispersive X-Ray Fluorescence (ED-XRF). The purpose is to provide the forensic scientist with the awareness of the value of debris on skin and then to highlight implications for forensic investigations. Results demonstrated that even naked eye observation can reveal useful trace materials, for defining the possible surrounding environment. As a next step, the episcopic microscope can increase the number of visible particulates and their analysis. In parallel, the ED-XRF spectroscopy can be useful to add a first chemical composition to the morphological data. Finally, the SEM-EDX analysis on small samples can provide the greatest morphological detail and the most complete chemical analysis, although limited, like the previous technique, to inorganic matrices. The analysis of debris on the skin, even with the difficulties due to the presence of contaminants, can provide information on the environments involved in criminal events that can add to the investigation framework.

Why identification matters: an explorative study on six cases of family reunification.

The escalating phenomenon of migration, accompanied by a disturbing surge in associated tragedies, has persistently violated internationally protected human rights. Absence of physical evidence, namely the presence of adequately identified corpses, may impede the full enjoyment of human rights and-in some cases-the course of justice as it obstructs the initiation of legal proceedings against individuals implicated in causing such catastrophes. It also presents administrative obstacles, as death certificates are indispensable in legitimizing statuses like orphanhood and widowhood. Family reunification, particularly for orphans, plays a significant role for those attempting to reconnect with their relatives all over the world. Likewise, for mothers, the acknowledgment of their marital status or widowhood can be a pathway to regain their marginalized right to social life. To elucidate this issue, we analyzed six representative cases from the tragic October 3, 2013, shipwreck near the Italian island of Lampedusa, where 366 individuals were retrieved dead from the sea. These cases underscore the practical challenges involved, highlighting the compelling need for continued efforts to ensure that this burdensome problem transcends from being a mere ethical, moral, and legal discourse. Although considerable progresses, these cases also reveal that substantial work still lies ahead. There is a pressing need for improved mechanisms to certify kinship ties, which are often the limiting factor in many reunifications, and can hinder the granting of custody to children. The severity and far-reaching implications of this problem necessitate thoughtful attention and action, especially considering the ongoing escalation in migration and related fatalities.

Digital pair-matching of iliac bones: pilot study on a three-dimensional approach with models acquired through stereophotogrammetry.

Three-dimensional (3D) pair-matching has brought about an innovative approach for the analysis of commingled skeletal remains, and it has been tested on bone models acquired through CT and laser scans. Here, 3D models of 40 innominate bones (20 left and 20 right) of 20 documented male individuals from a cemeterial skeletal collection were acquired through a stereophotogrammetric device (VECTRA M3, Canfield Scientific, Inc.). The ventral iliac surface was chosen as the anatomical region of interest (ROI) for the analysis. Each left ROI was mirrored and superimposed on the matching right ROI (contralateral element from the same individual) and mismatching ROIs (contralateral elements from different individuals). The point-to-point distance between models was calculated through the Vectra Analysis Module (VAM) software and the root mean square (RMS) point-to-point distance value was used to evaluate the sorting performance of the method, in terms of sensitivity and specificity rates. Differences in RMS between matches and mismatches were investigated through a Student's t test (p < 0.05). The state of preservation of the remains was assessed following an index of anatomical completeness and differences in RMS distances of true matches according to different anatomical completeness were assessed through the Mann-Whitney test (p < 0.05). RMS point-to-point distances of matches and mismatches were significantly different (p < 0.01), being the matches lower than mismatches. The RMS threshold of 2.9 mm identified all the true pairs; the test was 100% sensitive and 51% specific. The RMS of matches with a better state of preservation are significantly lower than the less preserved matches (p < 0.05). In general, a low RMS distance value may indicate a true match, being it to be further verified. The 3D approach for sorting innominate bones provides a valid screening test that could complete subjective and osteometric methods with numerical evidence of the match. Preliminary data suggest a possible relation between RMS distance values and taphonomic condition, which would benefit from further research.

The rights of migrants to the identification of their dead: an attempt at an identification strategy from Italy.

Europe is turning a blind eye on a humanitarian disaster unfolding at its doorsteps, with thousands of migrants dying unidentified in Mediterranean waters. Since 2014, Italy has been struggling in an almost indifferent international scenario to identify its dead migrants. Despite the lack of sufficient resources, of the difficulties in collecting post mortem data from the disseminated bodies, and of the problems of contacting and collecting ante mortem information from relatives, it has been proven, with a series of pilot studies, that not only can these bodies be identified but that relatives are also looking for their loved ones and need death certificates. This article focuses on the administrative limbo and lack of regulations obliging single states to engage in appropriate procedures to maximise identification.

Identification issues in bodies maimed by train fatalities: The role of the forensic pathologist.

Traffic accident injuries are relevant causes of mortality in industrialized countries. Among traffic accidents, the most destructive effect on the victim's body is produced by trains, which in a very short time provoke a complete distortion of the normal anatomical profiles and extensive amputations of the body. Deaths by severe body mutilation and/or decapitation require complex forensic investigation, as the victims' identification and the recognition of their body are not often feasible. In this study, a retrospective analysis of a series of cases of railway accidents is presented, with the aim of showing potential identification strategies, including a systematic physical re-assembling of the body. Among the railway fatalities assessed at the University Institute of Legal Medicine of Milan from 2016 until 2020, 61 cases were analyzed, focusing on the collection of postmortem data to improve accuracy for the identification of the victims. The results showed that the identification could be based on elements traditionally called "secondary identifiers" which, however, are becoming more and more important for positive identification, especially when dealing with migrants, vagrants, and homeless people. However, several issues need to be considered such as the limitations of the public prosecutors to perform expensive analyses (i.e., genetic testing), and the lack of antemortem data to be compared with postmortem findings, both odontological and fingerprint. In cases where it is not possible to proceed with identification or formal recognition of victims of railroad accidents with severe mutilations, a systematic physical re-assembling of the body is critical to reaching positive identification.

Two peculiar cases of cranial fractures running through craniotomy burr holes: may this be a kind of "exception" to the Puppe rule.

A correct assessment on the position, path, and direction of fracture lines is crucial when the sequence of different injuries on the skull has to be ascertained. In this context, the so-called Puppe's rule on intersecting fracture lines has always been considered a cornerstone of such an investigation. However, there is one factor that has never been previously considered: how do fracture lines behave when they reach the edges of an old and remodeled hole from a previous craniotomy? Two peculiar cases are presented of subjects undergoing cranial fractures due to blunt force trauma (case 1) and gunshot (case 2). Both previously underwent neurosurgical operations with persistence of the burr holes produced by the craniotomy drill (15 and 20 years before death). What was arguable, according to Puppe's rule, was that the fracture lines, when at the edge of the craniotomy hole, stopped. However, what has been detected was different than what expected: fracture lines continued exactly in the opposite direction, as though they were "skipping" the hole, following the same direction and the same axis and stopping a few centimeters over on the opposite side of the craniotomy hole. Puppe's rule has never been refuted, but these cases are the closest to an exception ever seen in forensic anthropology.

Taphonomic study on drowned victims in a non-sequestered aquatic environment in the Mediterranean Sea.

Human decomposition in sea water poses several challenges to forensic practitioners tasked with the analysis of drowned bodies. Postmortem changes in the marine environment have not been extensively investigated and the current literature is mainly based on bodies retrieved from shallow waters or on limited samples. On 18 April 2015, a fishing boat carrying allegedly 1,000 migrants sank in the Mediterranean Sea. In a fifteen-month time span, humanitarian missions were carried out to recover the bodies from the sea. The present study investigates postmortem changes on the drowned victims in a non-sequestered environment in the Mediterranean Sea. A retrospective study was performed by two observers on the autopsy photographic records in the series of bodies recovered from the open sea. For 184 bodies, the postmortem changes were evaluated according to facial, body, limb, and total aquatic decomposition score (FADS, BADS, LADS, TADS, respectively). Furthermore, a modification to the current scoring system that divides upper and lower limbs was suggested. The interobserver agreement was assessed using Krippendorff's alpha coefficient. Possible relations between the decomposition scores and PMSI were investigated with Pearson correlation analysis. According to the sequence of the recovery missions, increasing FADS, BADS, LADS, and TADS were observed. The modified scoring system showed a strong agreement between observers, allowing a more accurate description of the actual extent of decomposition. The scores showed a significant relation with the PMSI (p < 0.01). Prolonged submersion in the open marine environment was confirmed to show increasing decomposition rates, from moderate decay to full disarticulation. This study provides a descriptive unicum of the postmortem changes in the open sea, which may contribute to strengthen the discipline and aid description of bodies recovered in similar circumstances, especially if a body needs to be associated to a disaster or period with respect to another, thus facilitating families or authorities in the search for specific victims.

Bone tissue preservation in seawater environment: a preliminary comparative analysis of bones with different post-mortem histories through anthropological and radiological perspectives.

Bone taphonomy is a widely investigated topic; however, few data are available concerning marine bone taphonomy, especially on remains recovered from great depths and with short post-mortem intervals. To date, few studies have evaluated the bony changes which occur in seawater compared to samples with different post-mortem histories, and none through a comparative analysis of different approaches. To this purpose, this pilot study aims to examine the influence of seawater on bone preservation compared to other depositional contexts by multiple perspectives. Forty-nine human bone samples (femurs or tibiae) recovered from different environments (sea water, fresh water, outdoor, burial in coffin) were compared by macroscopic, microscopic and bone densitometric approaches. In order to investigate organic and inorganic components, undecalcified and decalcified histology of thin sections was performed. The analyses revealed a well-preserved bone tissue both macroscopically (92%) and microscopically (97% and 95% for undecalcified and decalcified sections). No significant differences were detected from radiological densitometric investigations (BMD = 1.6 g/cm2 ± 0.1), except between old and young individuals (p value < 0.001). Differences were observed for body decomposition and few scavenged samples (3/15). However, even if slight variations were observed, no relation was recorded with the depositional contexts. We found a similar bone preservation in the four environments at the time of recovery, both macroscopically and microscopically, but also with radiological densitometric investigations. Our observations enriched the literature on bone taphonomy, providing data on bone tissue preservation in the early post-mortem period from a multidisciplinary perspective, paving the way for further studies on the topic.

Twenty-five years of unidentified bodies: an account from Milano, Italy.

The identification of decedents has always been a central issue in forensic pathology, for ethical, criminal, and administrative reasons, but today, it needs more attention due to issues related not only to migration but also to the weakening of family ties. This article presents a descriptive study discussing the Italian regulatory situation developed in the last decade to face the many identification issues, with all its improvements and flaws. Hence, data gathered in 25 years of at the Institute of Legal Medicine of Milan and the epidemiology of unidentified decedents are illustrated. Briefly, from 1995 to 2019, the number of unidentified human remains with no identity or requesting verification of identity amounts to 726, i.e., 3% circa of all autopsies performed at the Institute, with an average of 29 individuals per year. In total, 528 (72.7%) individuals were successfully identified, 100 (13.8%) remained without an identity, and 98 (13.5%) individuals remained with suspected yet unconfirmed identities. Percentages for each identification technique are displayed, with insight into the role of forensic anthropology and odontology compared to genetics, and into the misuse of non-scientific methods allowed by Public Prosecutors. All the data is compared, as much as possible, with the very few recent studies concerning the problem worldwide. Finally, the article aims to show the Italian experience in dealing with unidentified bodies, in order to provide food for thought for other countries toward a discussion regarding a global issue which is sometimes taken for granted and underestimated.

Common and much less common scenarios in which botany is crucial for forensic pathologist and anthropologists: a series of eight case studies.

It is commonly accepted that crime scene recovery and recording are key moments of any judicial inspection in which investigators must decide on the correct strategies to put into place. Complex outdoor scenarios, presenting partially or entirely skeletonised remains, can benefit more than others by the intervention of environmental specialists (forensic anthropologists, archaeologists, entomologists and botanists). These experts are capable of singling out, correctly recording and recovering environmental evidence that can lead to a more comprehensive reconstruction of a given criminal episode. If human remains are discovered in an outdoor scenario, the on-site presence of a botanist will guarantee a correct approach to the identification, recording and recovery of any botanical evidence. If an on-site botanist is not available, the operators must be capable of both the botanical evaluation of a scene and the implementation of correct botanical sampling protocols.The following collection of unusual case histories that aim at underlining the efficacy of forensic botany will examine the determination of post mortem or the post depositional interval, evidence for a victim's post mortem transfer, evidence for the identification of a primary crime scene and evidence for the identification of a victim's dismemberment site. In another two cases, one, we will illustrate the important role that forensic botany played in the discrimination between botanical material used to voluntarily conceal a victim and vegetation that had grown naturally above a disposal site, whereas the other will highlight the protocols implemented for the identification of a murder weapon.

Three-dimensional analysis of sphenoid sinus uniqueness for assessing personal identification: a novel method based on 3D-3D superimposition.

Sphenoid sinuses are considered the most variable structures of human body: therefore, they may be used for personal identification, through the application of 3D segmentation procedures. This study aims at proposing a new protocol for personal identification based on 3D-3D superimposition of sphenoid sinuses segmented from head CT scans. Adult subjects (equally divided among males and females) who underwent two head CT scans were extracted from a hospital database. Sphenoid sinuses were segmented through ITK-SNAP software and the corresponding 3D models were automatically superimposed to obtain 40 matches (when they belonged to the same person) and 260 mismatches (when they were extracted from different individuals). The RMS (root mean square) point-to-point distance was then calculated for all the superimpositions: differences according to sex and group (matches and mismatches) were assessed through the Mann-Whitney test (p < 0.05). On average, the RMS value was almost ten times smaller in matches (0.22 ± 0.11 mm) than in mismatches (2.16 ± 0.57 mm) with a statistically significant difference according to group (p < 0.05), but not to sex (p > 0.05). The study proposed a new method for assessing personal identification from segmented 3D models of sphenoid sinuses, useful in the forensic contexts where other methods might not be implementable or successful.

The comparative performance of PMI estimation in skeletal remains by three methods (C-14, luminol test and OHI): analysis of 20 cases.

When estimating post-mortem interval (PMI) in forensic anthropology, the only method able to give an unambiguous result is the analysis of C-14, although the procedure is expensive. Other methods, such as luminol tests and histological analysis, can be performed as preliminary investigations and may allow the operators to gain a preliminary indication concerning PMI, but they lack scientific verification, although luminol testing has been somewhat more accredited in the past few years. Such methods in fact may provide some help as they are inexpensive and can give a fast response, especially in the phase of preliminary investigations. In this study, 20 court cases of human skeletonized remains were dated by the C-14 method. For two cases, results were chronologically set after the 1950s; for one case, the analysis was not possible technically. The remaining 17 cases showed an archaeological or historical collocation. The same bone samples were also screened with histological examination and with the luminol test. Results showed that only four cases gave a positivity to luminol and a high Oxford Histology Index (OHI) score at the same time: among these, two cases were dated as recent by the radiocarbon analysis. Thus, only two false-positive results were given by the combination of these methods and no false negatives. Thus, the combination of two qualitative methods (luminol test and microscopic analysis) may represent a promising solution to cases where many fragments need to be quickly tested.

Luminol testing in detecting modern human skeletal remains: a test on different types of bone tissue and a caveat for PMI interpretation.

When forensic pathologists and anthropologists have to deal with the evaluation of the post-mortem interval (PMI) in skeletal remains, luminol testing is frequently performed as a preliminary screening method. However, the repeatability of this test on the same bone, as well as comparative studies on different bones of the same individual, has never been performed. Therefore, with the aim of investigating the influence that different types of bones may exert on the response to the luminol test, the present study analysed three different skeletal elements (femoral diaphysis, vertebra and cranial vault), gathered from ten recent exhumed skeletons (all with a 20-year PMI). The analysis was performed twice on the same bone after 2 months: the analysis at time 0 concerned the whole bone, whereas the second concerned only a part of the same bone taken during the first test (which already had been broken). The overall results showed different responses, depending on the type of bone and on the integrity of the samples. Negative results at the first analysis (6.6% out of the total of samples) are consistent with what is reported in the literature, whilst at the second analysis, the increase of about 20% of false-negative results highlights that the luminol test ought to be performed with caution in case of broken bones or elements which are taphonomically altered. Results have thus proven that the exposition to environmental agents might result in haemoglobin (Hb) loss, as detected even after only 2 months. The study also focused on the crucial issue of the type of bone subjected to testing, remarking the suitability of the femoral diaphysis (100% of positive responses at the first analysis vs only 18% of false-negative results at the second test, corresponding to 5% of total false-negative results) as opposed to other bone elements that showed a low yield. In particular, the cranial vault gave poor results, with 40% of discrepancy between results from the two analyses, which suggests caution in choosing the type of bone sample to test. In conclusion, luminol testing should be used with caution on bones different from long bones or on non-intact bones.

Analysis of Cutmarks on Bone: Can Light Microscopy Be of Any Help?

One of the main issues in forensic anthropology consists of the identification of signs of trauma in skeletal remains, including sharp-force injuries. So far, several studies have been performed to assess differences between injuries caused by different instruments, not, however, through light microscopy.In this study, 152 sharp-force injuries were performed by 5 different tools through 2 different orientations on 2 humeral diaphyses and were analyzed by stereo and light microscopy to assess possible morphological differences.This study showed that although W-shaped injuries are frequently reported in cases of wood-cutting saws, other shapes are often observed; lesions due to metal-cutting saws are almost always U shaped, whereas injuries caused by knives are V shaped. Although cut marks may represent a variable range of features, the present study was able to highlight typical profiles that may be of some help for the diagnosis of weapon and the intentionality of the action.

Characteristics and time-dependence of cut marks and blunt force fractures on costal cartilages: an experimental study.

PURPOSE: The distinction between cut marks and blunt force injuries on costal cartilages is a crucial issue in the forensic field. Moreover, a correct distinction may further be complicated by decomposition, so the need arises to investigate the distinctive features of lesions on cartilage and their changes over time. METHODS: This study aimed to assess the stereomicroscopic features of cut marks (performed with six different knives) and blunt fractures (performed with a hammer and by means of manual bending) on 48 fragments of human costal cartilages. Moreover, in order to simulate decomposition, the cut and fractured surfaces were checked with stereomicroscopy and through casts after 1 and 2 days, 1 week, and 1, 2 and 4 months of drying in ambient air. RESULTS: In fresh samples, for single and unique cuts, striations were observed in between 44 and 88% of cases when non-serrated blades were used, and between 77 and 88% for serrated blades; in the case of "repeated" (back and forth movement) cuts, striations were detected in between 56 and 89% of cases for non-serrated blades, and between 66 and 100% for serrated blades. After only 1 week of decomposition the detection rates fell to percentages of between 28 and 39% for serrated blades and between 17 and 33% for non-serrated blades. Blunt force injuries showed non-specific characteristics, which, if properly assessed, may lead to a reliable distinction between different cut marks in fresh samples. The most evident alterations of the structure of the cartilage occurred in the first week of decomposition in ambient air. After one week of drying, the characteristics of cut marks were almost undetectable, thereby making it extremely challenging to distinguish between cut marks, blunt force fractures and taphonomic effects. CONCLUSION: The study represents a contribution to the correct assessment and distinction of cut marks and blunt force injuries on cartilages, providing a glimpse on the modifications such lesions may undergo with decomposition.

Assets and pitfalls of chemical and microscopic analyses on gunshot residues in skeletonized bodies: a report of five cases.

In case of gunshot wounds, forensic anthropologists and pathologists have many tools at hand, and the assistance that chemical and microscopic investigations can provide in such scenarios is often valuable and crucial. However, the results of such analyses in the search of gunshot residues (GSR) ought not to be acritically considered. We report five cases where chemical (sodium rhodizonate) and microscopic (scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDX)) analyses were performed for the search of GSR. Four cases concerned the forensic field and analyses on buried, charred, or submerged remains, whereas one case concerned the historical remains of a soldier of the First World War. In every case, the search for GSR with these techniques showed their persistence even after long periods and preservation in peculiar environments. However, chemical analyses provided their contribution, but in two cases, anthropological analyses provided crucial and solving results. The five cases show the indisputable usefulness of chemical and microscopic analyses in the search of GSR in gunshot wounds and especially how such residues may survive in time and in adverse environmental conditions. However, experts should always be dubious about some pitfalls (such as contamination) one can frequently find in these scenarios.

Splitting hairs: differentiating between entomological activity, taphonomy, and sharp force trauma on hair.

PURPOSE: The analysis of hair can provide useful information for the correct evaluation of forensic cases, but studies of trauma on hair are extremely rare. Hair may present lesions caused by traumatic events or by animals: in fact, signs of sharp force weapons on hair may provide important information for the reconstruction of the manner of death, and, for example, may suggest fetishist practice. This study stemmed from a judicial case where it was fundamental to distinguish between sharp force lesions and insect activity on hair. METHODS: In order to highlight differences between sharp force lesions and insect feeding activity, different experiments were performed with high power microscopy: hair samples were subjected to several lesions by blunt and sharp force trauma; then samples were used as pabulum for two taxa of insects: the common clothes moth (Tineola bisselliella Lepidoptera, Tineidae) and the carpet beetle (Anthrenus sp., Coleoptera, Dermestidae). Hairs were examined from a macroscopic and microscopic point of view by stereomicroscopy and scanning electron microscopy (SEM): the morphological characteristics of the lesions obtained from the different experimental samples were compared. RESULTS: Results show that sharp force trauma produces lesions with regular edges, whereas insects leave concave lesions caused by their "gnawing" activity. These two types of lesions are easily distinguishable from breaking and tearing using SEM. CONCLUSIONS: This study demonstrates that insect activity leaves very specific indications on hair and sheds some light on different hair lesions that may be found in forensic cases.

The potential of facial nevi in personal identification.

Forensic anthropologists dealing with personal identification (PI) of human remains have recently stressed the need to explore the potential of "secondary identifiers" for identifying victims who died in particular events for whom images often represent the main antemortem data available. Being the face the part most exposed in images, characteristics as pigmented skin lesions (PSLs), can be crucial if combined with other input. Since no data is available on frequencies and distribution of facial PSLs in the general population, this study aims at systematically collecting such data to verify their potential in PI and to open a debate on the aid that "secondary identifiers", regardless of their specific nature, can give to the identification of the deceased in specific forensic contexts. A retrospective analysis on three-dimensional facial models of 1039 Italian subjects (from 4 to 84 years old) was conducted to examine the incidence of PSLs discriminated according to size and position in well-defined facial areas. From the collected data we developed a probabilistic approach providing the likelihood ratio (LR) for two settings: (1) the relative frequencies of nevi in the various facial areas, providing the deriving compound probability of owning a certain facial PSLs pattern; and (2) codes describing the facial nevi pattern of each individual of our population, thus testing their uniqueness and so their potential in PI. The calculated LRs mostly proved high identifying strength, particularly when provided by the compound probability-based approach. Data on incidence and position of facial nevi, their generated codes, and the probabilistic approach here presented, all constitute a starting point for advancing secondary identifiers. Nonetheless, although this preliminary study proved facial PSLs as valuable and potentially useful for identification, their significance and validity should be interpreted with caution as we are still at the first theoretical step clearly based on ideal conditions, and thus further investigations are due on the limitations of their use in practical identifying settings. Therefore, being this systematic study only a preliminary one in its nature, it is recommended not to use this kind of approach until further studies will test its validity in several practical conditions.