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We review the application of Nuclear Analytical Techniques (NATs) to forensic problems for the first time. NATs include neutron activation analysis (NAA), carried out in nuclear reactors for elemental analysis; accelerator-based techniques, mainly Ion Beam Analysis (IBA) for elemental and molecular analysis; and Accelerator Mass Spectrometry (AMS) for dating of traces of forensic interest by "radiocarbon dating" and other related methods. Applications include analysis of drugs of abuse, food fraud, counterfeit medicine, gunshot residue, glass fragments, forgery of art objects and documents, and human material. In some applications only the NATs are able to provide relevant information for forensic purposes. This review not only includes a wide collection of forensic applications, but also illustrates the wide availability worldwide of NATs, opening up opportunities for an increased use of NATs in routine forensic casework., Competing Interests: Declaration of Competing Interest The authors declare they have no conflicts of interest., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

While plant species identification in forensics can be useful in cases involving poisonous, psychoactive, or endangered plant species, it can also become quite challenging, especially, when dealing with processed, decaying, colonized or infected material of plant origin. The Animal Plant and Soil Traces expert working group of the European Network of Forensic Science Institutes in their best practice manual has recommended several markers for plant species identification. Current study is a part of implementation of method in a forensic laboratory and its aim is to evaluate four of the recommended markers (ITS, matK, rbcL, and trnH-psbA) for species identification of forensically important plant species including medicinal, poisonous, psychoactive, and other plants. Such parameters as PCR and sequencing success, sequence length, species resolution rate and species cover in GenBank were analysed. Blind testing was performed to evaluate use of the markers for identification of forensically more complicated samples. According to results, a combination of ITS, matK and trnH-psbA is the best choice for plant species identification. The best results with fresh plant material can be achieved with ITS, trnH-psbA, and matK, while ITS and matK are the best choice when working with low quality plant material. rbcL due to its low species discrimination rate can be used only as an indicative marker., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Refractive errors (RE) are commonly reported visual impairment problems worldwide. Previous clinical studies demonstrated age-related changes in human eyes. We hypothesized that the binocular RE metrics including sphere and cylinder power, axis orientation, and interpupillary distance (IPD) can be used for forensic age estimation of an unknown individual. RE data of both eyes were collected from the clinical optometric exams and prescription glasses of 2027 Egyptian individuals aged between 2 to 93 years. The differences between age groups as well as sides, and sexual dimorphism were explored. Two modeling methods were compared: multiple and stepwise linear regression (LR) versus machine learning Regression Forest (RFM). Data were apportioned into training and test datasets with a ratio of 80/20. The results showed significant differences among the age groups in each eye for all variables. Stepwise LR improved the results over models based on the one-sided lens due to selection of IPD in addition to the left and right axis, and left sphere as independent variables. For the RFM, the left axis and IPD were the most important features. RFM outperformed LR in terms of accuracy and root mean squared error (RMSE). The estimated age within ±10 years showed 81.4% accuracy rate and RMSE = 8.9 years versus 38.5% accuracy rate and RMSE = 17.99 years using RFM and stepwise LR, respectively, in the test set. The current study upholds the significance of the age-related changes of refractive error in formulating alternative forensic age estimation models when standard methods are unavailable., (© 2024 American Academy of Forensic Sciences.)

Biological material is routinely collected at crime scenes and from exhibits and is a key type of evidence during criminal investigations. Improvements in DNA technologies allow collection and profiling of trace samples, comprised of few cells, significantly expanding the types of exhibits targeted for DNA analysis to include touched surfaces. However, success rates from trace and touch DNA samples tend to be poorer compared to other biological materials such as blood. Simultaneously, there have been recent advances in the utility of environmental DNA collection (eDNA) in identification and tracking of different biological organisms and species from bacteria to naked mole rats in different environments, including, soil, ice, snow, air and aquatic. This paper examines the emerging methods and research into eDNA collection, with a special emphasis on the potential forensic applications of human DNA collection from air including challenges and further studies required to progress implementation., (© 2024 The Authors. ELECTROPHORESIS published by Wiley‐VCH GmbH.)

iPhone operating system (iOS) devices utilize binary cookies as a data storage tool, encoding user-specific information within an often-neglected element of smartphone analysis. This binary format contains details such as cookie flags, expiration, and creation dates, domain, and value of the cookie. These data are invaluable for forensic investigations. This study presents a comprehensive methodology to decode and extract valuable data from these files, enhancing the ability to recover user activity information from iOS devices. This paper provides an in-depth forensic investigation into the structure and function of iOS binary cookie files. Our proposed forensic technique includes a combination of reverse engineering and custom-built Python scripts to decode the binary structure. The results of our research demonstrate that these cookie files can reveal an array of important digital traces, including user preferences, visited websites, and timestamps of online activities. It concludes that the forensic analysis of iOS binary cookie files can be a tool for forensic investigators and cybersecurity professionals. In the rapidly evolving domain of digital forensics, this research contributes to our understanding of less-explored data sources within iOS devices and their potential value in investigative contexts., (© 2024 American Academy of Forensic Sciences.)

Tire impression evidence can be a valuable tool during a crime scene investigation-it can link vehicles to scenes or secondary locations, and reveal information about the series of events surrounding a crime. The interpretation of tire impression evidence relies on the expertise of forensic tire examiners. To date, there have not been any studies published that empirically evaluate the accuracy and reproducibility of decisions made by tire impression examiners. This paper presents the results of a study in which 17 tire impression examiners and trainees conducted 238 comparisons on 77 distinct questioned impression-known tire comparison sets (QKsets). This study was conducted digitally and addressed examinations based solely upon the characteristics of the tire impression images provided. The quality and characteristics of the impressions were selected to be broadly representative of those encountered in casework. Participants reported their decisions using a multi-level conclusion scale: 68% of responses were class associations (Association of Class Characteristics or Limited Association of Class), 21% were definitive decisions (ID or Exclusion), 8% were probable decisions (High Degree of Association or Indications of Non-Association), and 3% were neutral responses (Not Suitable or Inconclusive). Although class associations were the most reported response type, when definitive decisions were reported, they were often correct: 96% of IDs and 89% of Exclusions were consistent with ground truth regarding the source of the known tire in the QKset. Overall, we observed 4 erroneous definitive decisions (3 Exclusions on mated QKsets; 1 ID on a nonmated QKset) and 1 incorrect probable decision (Indications of Non-Association on a mated QKset). Decision rates were notably associated with both quality (lower quality questioned impressions were more likely to result in class associations) and dimensionality (2D questioned impressions were more likely to result in definitive decisions), which were correlated factors. Although the study size limits the precision of the measured rates, the results of this study remain valuable to the forensic science and legal communities and provide empirical data regarding examiner performance for a discipline that previously did not have any such estimates., Competing Interests: Declaration of Competing Interest Authors declare no competing interests., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Hair is a commonly encountered trace evidence in wildlife crimes involving mammals and can be used for species identification which is essential for subsequent judicial proceedings. This proof of concept study aims, to distinguish the black guard hair of three wild cat species belonging to the genus Panthera i.e. Royal Bengal Tiger (Panthera tigris tigris), Indian Leopard (Panthera pardus fusca), and Snow Leopard (Panthera uncia) using a rapid and non-destructive ATR-FTIR spectroscopic technique in combination with chemometrics. A training dataset including 72 black guard hair samples of three species (24 samples from each species) was used to construct chemometric models. A PLS2-DA model successfully classified these three species into distinct classes with R-Square values of 0.9985 (calibration) and 0.8989 (validation). VIP score was also computed, and a new PLS2DA-V model was constructed using variables with a VIP score ≥ 1. External validation was performed using a validation dataset including 18 black guard hair samples (6 samples per species) to validate the constructed PLS2-DA model. It was observed that PLS2-DA model provides greater accuracy and precision compared to the PLS2DA-V model during cross-validation and external validation. The developed PLS2-DA model was also successful in differentiating human and non-human hair with R-Square values of 0.99 and 0.91 for calibration and validation, respectively. Apart from this, a blind test was also carried out using 10 unknown hair samples which were correctly classified into their respective classes providing 100 % accuracy. This study highlights the advantages of ATR-FTIR spectroscopy associated with PLS-DA for differentiation and identification of the Royal Bengal Tiger, Indian Leopard, and Snow Leopard hairs in a rapid, accurate, eco-friendly, and non-destructive way., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Chartered Society of Forensic Sciences. Published by Elsevier B.V. All rights reserved.)

Age estimation is a critical aspect of reconstructing a biological profile in forensic sciences. Diverse biochemical processes have been studied in their correlation with age, and the results have driven DNA methylation to the forefront as a promising biomarker. DNA methylation, an epigenetic modification, has been extensively studied in recent years for developing age estimation models in criminalistics and forensic anthropology. Epigenetic clocks, which analyze DNA sites undergoing hypermethylation or hypomethylation as individuals age, have paved the way for improved prediction models. A wide range of biomarkers and methods for DNA methylation analysis have been proposed, achieving different accuracies across samples and cell types. This review extensively explores literature from the past 5 years, showing scientific efforts toward the ultimate goal: applying age prediction models to assist in human identification., Competing Interests: The authors declare no conflicts of interest.

Molecular physics plays a pivotal role in various fields, including medicine, pharmaceuticals, and broader industrial applications. This study aims to enhance the methods for producing specific optically active materials with distinct spectroscopic properties at the molecular level, which are crucial for these sectors, while prioritizing human safety in both production and application. Forensic science, a significant socio-economic field, often employs hazardous substances in analyzing friction ridges on porous surfaces, posing safety concerns. In response, we formulated novel, non-toxic procedures for examining paper evidence, particularly thermal papers. Our laboratory model utilizes a polyvinyl alcohol polymer as a rigid matrix to emulate the thermal paper's environment, enabling precise control over the spectroscopic characteristics of 1,8-diazafluoro-9-one (DFO). We identified and analyzed the cyclodimer 1,8-diazafluoren-9-one (DAK DFO), which is a non-toxic and biocompatible alternative for revealing forensic marks. The reagents used to preserve fingerprints were optimized for their effectiveness and stability. Using stationary absorption and emission spectroscopy, along with time-resolved emission studies, we verified the spectroscopic attributes of the new structures under deliberate aggregation conditions. Raman spectroscopy and quantum mechanical computations substantiated the cyclodimer's configuration. The investigation provides robust scientific endorsement for the novel compound and its structural diversity, influenced by the solvatochromic sensitivity of the DFO precursor. Our approach to monitoring aggregation processes signifies a substantial shift in synthetic research paradigms, leveraging simple chemistry to yield an innovative contribution to forensic science methodologies.

The polymerase chain reaction (PCR) has played a fundamental role in our understanding of the world, and has applications across a broad range of disciplines. The introduction of PCR into forensic science marked the beginning of a new era of DNA profiling. This era has pushed PCR to its limits and allowed genetic data to be generated from trace DNA. Trace samples contain very small amounts of degraded DNA associated with inhibitory compounds and ions. Despite significant development in the PCR process since it was first introduced, the challenges of profiling inhibited and degraded samples remain. This review examines the evolution of the PCR from its inception in the 1980s, through to its current application in forensic science. The driving factors behind PCR evolution for DNA profiling are discussed along with a critical comparison of cycling conditions used in commercial PCR kits. Newer PCR methods that are currently used in forensic practice and beyond are examined, and possible future directions of PCR for DNA profiling are evaluated.

This article investigates a critical part of professional careers: adhering to a code of ethics that defines proper behavior within the context of one's specialized practice. It aims to improve the setting of forensic entomology work by highlighting the ethical issues that frequently emerge. The primary goal is to uncover common ethical issues involved in this field's everyday activities and provide informed advice on appropriate resolution options. This investigation dives into the complex interaction of religious beliefs, cultural aspects, and issues of ethics, evaluating their relevance in forensic entomology. It examines a wide range of ethical issues, from the ethics involved in collecting samples at crime scenes and morguesto the ethics created into scientific investigations, report writing, and court testimony. These guiding principles are critical in setting scientific standards and building public trust in forensic entomology's ability to provide exceptional services. Notably, regardless of whether the forensic entomology service provider is accredited by any validating organization, these ethical requirements remain critical, emphasizing their global application., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Chartered Society of Forensic Sciences. Published by Elsevier B.V. All rights reserved.)

In Colombia there are estimated to be over 121,000 missing people and victims of forced disappearances. Forensic investigators therefore need assistance in determining optimal detection techniques for buried victims, to give victims' families closure and for the wider community to see that justice is being served. Previous research has created 12 controlled simulated clandestine graves of typical Colombian murder victim scenarios at 0.5 m - 1.2 m depths in savannah and rainforest sites in Colombia. The 0-3 years of geophysical monitoring results of were published, with this paper reporting on 4-8 years monitoring of both UAV drone results and geophysical data. The UAV results from the year 8 survey, published for first time from Colombia, showed that the simulated graves could still be located using NDVI and NIR multi-spectral data, but not using optical or other multi-spectral data. The 0-3 years of geophysical data found the simulated clandestine graves could be detected with electrical resistivity and GPR methods, with the 4-8 year surveys evidencing that they could still be detected using bulk ground conductivity surveys, GPR horizontal time slice datasets and 2D ERT profiles. Research implications suggest initial use of UAV remote sensing technology to pinpoint likely search areas, before subsequent ground reconnaissance, geophysical surveys and their interpretation, before intrusive investigation methods are employed for detecting missing and disappeared persons in Colombia., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Research in many forensic science fields commonly uses domestic pigs (Sus spp.) as proxies for human remains, due to their physiological and anatomical similarities, as well as being more readily available. Unfortunately, previous research, especially that which compares the decompositional process, has shown that pigs are not appropriate proxies for humans. To date, there has not been any published research that specifically addresses whether domestic pigs are adequate human proxies for the geophysical detection of clandestine graves. As such, the aim of this paper was to compare the geophysical responses of pig cadavers and human donor graves, in order to determine if pigs can indeed be used as adequate human proxies. To accomplish this, ground penetrating radar (GPR) and electrical resistivity tomography (ERT) responses on single and multiple pig cadaver graves were compared to single and multiple human donor graves, all of which are in known locations within the same geological environment. The results showed that under field conditions, both GPR and ERT were successful at observing human and pig burials, with no obvious differences between the detected geophysical responses. The results also showed that there were no differences in the geophysical responses of those who were clothed and unclothed. The similarity of the responses may reflect that the geophysical techniques can detect graves despite what their contents are. The study implications suggest that experimental studies in other soil and climate conditions can be easily replicated, benefiting law enforcement with missing persons cases., (© 2023 The Authors. Journal of Forensic Sciences published by Wiley Periodicals LLC on behalf of American Academy of Forensic Sciences.)

Forensic facial image comparison based on recognition algorithms has been widely applied in forensic science. Previous researches have been concentrating on the cases of using single system during comparison, while how to use multiple systems has not yet been studied. In this paper, a dual-systems model (including SeetaFace and FaceNet) for facial comparison was constructed, and Bayesian networks were utilized as the basic frame. In order to prove its superiority, a large-scale experiment (on the dataset CelebA) has been carried on to evaluate the score-based likelihood ratio. We used three likelihood ratio evaluation tools (Empirical Cross-Entropy, Cost Likelihood Ratio, Limit Tippett Plots) to assess the performance of the model. The Wasserstein distance was also used to evaluate the detailed likelihood ratio performance. The experimental results show that the likelihood ratio performance of our dual-systems model is better than single system. Besides, our method of model building and evaluation can also be used in the condition of triple or more systems., Competing Interests: Declaration of Competing Interest All authors disclosed no relevant relationships., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Background: Bodily fluid stains are one of the most relevant evidence that can be found at the crime scene as it provides a wealth of information to the investigators. They help to report on the individuals involved in the crime, to check alibis, or to determine the type of crime that has been committed. They appear as stains in different types of substrates, some of them porous, which can interfere in the analysis. The spectroscopy techniques combined with chemometrics are showing increasing potential for their use in the analysis of such samples due to them being fast, sensitive, and non-destructive., Findings: This is a comprehensive review of the studies that used different spectroscopic techniques followed by chemometrics for analysing biological fluid stains on several surfaces, and under various conditions. It focuses on the bodily fluid stains and the most suitable spectroscopic techniques to study forensic scientific problems such as the substrate's characteristics, the influence of ambient conditions, the aging process of the bodily fluids, the presence of animal bodily fluids and non-biological fluids (interfering substances), and the bodily fluid mixtures. The most widely used techniques were Raman spectroscopy and attenuated total reflection Fourier transform infrared spectroscopy (ATR FTIR). Nonetheless, other non-destructive techniques have been also used, like near infrared hyperspectral imaging (HSI-NIR) or surface enhanced Raman spectroscopy (SERS), among others. This work provides the criteria for the selection of the most promising non-destructive techniques for the effective in situ detection of biological fluid stains at crime scene investigations., Significance and Novelty: The use of the proper spectroscopic and chemometric approaches on the crime scene is expected to improve the support of forensic sciences to criminal investigations. Evidence may be analysed in a non-destructive manner and kept intact for further analysis. They will also speed up forensic investigations by allowing the selection of relevant samples from occupational ones., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

With the advent of the post-genome era, omics technologies have developed rapidly and are widely used, including in genomics, transcriptomics, proteomics, metabolomics, and microbiome research. These omics techniques are often based on comprehensive and systematic analysis of biological samples using high-throughput analysis methods and bioinformatics, to provide new insights into biological phenomena. Currently, omics techniques are gradually being applied to forensic entomology research and are useful in species identification, phylogenetics, screening for developmentally relevant differentially expressed genes, and the interpretation of behavioral characteristics of forensic-related species at the genetic level. These all provide valuable information for estimating the postmortem interval (PMI). This review mainly discusses the available omics techniques, summarizes the application of omics techniques in forensic entomology, and their future in the field., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Wildlife forensics is defined as providing forensic evidence to support legal investigations involving wildlife crime, such as the trafficking and poaching of animals and/ or their goods. While wildlife forensics is an underexplored field of science, the ramifications of poaching can be catastrophic. The consequences of wildlife crime include disease spread, species and habitat loss, human injury, and cultural loss. Efforts to use forensic science to combat poaching are currently limited to DNA-based techniques. However, fingermark analysis for the identification of perpetrators of wildlife crimes has not been explored to the same extent, despite being a cost-effective, simple-to-use forensic method that is easy to deploy in-field. This review covers literature that has explored fingermark examination techniques used on wildlife-related samples, such as pangolin scales, ivory-based substances, bone, and eggs, as well as feathers and skins, among more obscure trafficked items. Useful preliminary work has been conducted in this subject area, demonstrating that commonly used fingermark analysis techniques can be applied to wildlife-based items. However, many of these studies suffer from limitations in terms of experimental design. More work should be done on creating studies with larger sample sizes and novel approaches should be validated under environmental conditions that mimic real crime scenes. Further research into determining the forensic fingermark analysis techniques that perform the most efficiently in the environmental conditions of the countries where they are needed would therefore benefit legal investigations and help to reduce instances of poaching., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Further steps toward understanding the time-related information contained within bloodstains found at the crime scene are rightly considered a top priority in forensic science. Contrary to widely held assumptions, the reason for the delayed exploitation of bloodstains dating methods in practice is not the lack of suitable analytical techniques for monitoring degradation processes. The problem lies in the variability of the environmental and circumstantial conditions, playing a vital role in the degradation kinetics of blood deposits. The present article demonstrates the possibility of breaking with current approaches based on absolute age estimations to finally answer time-centered questions in real forensic scenarios. The proposed novel framework for situating forensic traces in time is based on the likelihood ratio assessment of the (dis)similarity between the evidence decomposition and sets of reference materials obtained through supervised aging. In such a strategy, every dating procedure is constructed on a case-by-case basis to fit examined blood traces, thereby limiting the adverse influence of external factors on the validity of age estimations and providing a way for future crime scene implementation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Many different issues have been identified in forensic science for more than 10 years. While quality management has often been suggested as a path forward, research is generally considered as an essential part of the solution. Through an overview of current forensic science research, this paper aims at evaluating if and how research answer the challenges forensic science is currently facing. While forensic related publications have massively increased over the years, approximately half of the publications were published in non-forensic sources, indicating that forensic science research tends to be led by other disciplines. Over the years, forensic science research has remained largely oriented towards methodological and technological development rather than relevance to the forensic science discipline and practice. Practical implementation of the techniques is rarely discussed from a forensic perspective, and thus research rarely move from the "proof-of-concept" stage to its utilisation in case investigation. The digital transformation also generated a massive increase of data, making it challenging to find the relevant pieces of information in the mass of "forensic" publications available on-line. Thus, we propose to refocus forensic science research on forensic fundamental and practical questions to strengthen the discipline and its impact on crime investigation and security issues. Our propositions represent an incentive to further discuss forensic science research and knowledge transmission through the definition of a common culture within the community, focusing on common fundamental knowledge such as a better understanding of the concept of trace and its case-based information content., Competing Interests: Conflict of interests none., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Forensic Microbiology is an emerging branch of science that has great potential to assist criminal investigations. Having said that, microbial analysis is not performed routinely during forensic investigations in India. This could be attributed to lack of specific training and lack of evidence-based standard protocol., Objectives: The authors attempt to highlight the key areas in forensic microbiology that need to be explored in a developing nation like India., Content: Forensic microbiology could help in linking a person to a crime, determining the cause of death, estimating postmortem interval (PMI), etc. Additionally, applications are being developed by forensic microbiologists across the globe to investigate the coordinated and dynamic changes in microbial activity which occur after the death of a human host. Such evidence from the human postmortem microbiome can aid in criminal investigations and administration of justice. These recent advances and developments have the potential to transform the field of forensic microbiology in a developing country., Competing Interests: Declaration of competing interests The authors declare that there is no conflict of interest., (Copyright © 2023 Indian Association of Medical Microbiologists. Published by Elsevier B.V. All rights reserved.)

Microfluidic technology is a powerful tool to enable the rapid, accurate, and on-site analysis of forensically relevant evidence on a crime scene. This review paper provides a summary on the application of this technology in various forensic investigation fields spanning from forensic serology and human identification to discriminating and analyzing diverse classes of drugs and explosives. Each aspect is further explained by providing a short summary on general forensic workflow and investigations for body fluid identification as well as through the analysis of drugs and explosives. Microfluidic technology, including fabrication methodologies, materials, and working modules, are touched upon. Finally, the current shortcomings on the implementation of the microfluidic technology in the forensic field are discussed along with the future perspectives.

Ghosting is the phenomenon that exists when a footprint has a lighter area around the tip of one or more toes or a shadow-like area at the back of the heel. To date, ghosting has been considered primarily a finding of dynamic (walking) footprints, rather than static (standing) footprints. The prevalence of ghosting in static footprints is unknown, and research on its presence in static and dynamic footprints from the same participant is sparse, as are studies on its occurrence in different geographic populations. This study is among the first to evaluate the occurrence of ghosting in the static and dynamic footprints from a particular individual with participants in two geographic populations. A combination of both inkless and ink footprint collection systems were used to obtain a total of 206 bare footprints from 103 adult participants from the United States and India. The data comprised 103 static and 103 dynamic footprints. Ghosting occurred significantly in static footprints, though less frequently than in dynamic footprints. Ghosting in static footprints was seen most often at the first toe, followed by the third and second respectively. This aspect appeared least at the heel. In dynamic footprints, it occurred most at the first toe, followed by the second and fourth toes, and then the third toe and the heel. The prevalence of ghosting in footprints from the United States and India differed in their locations, notably at the first and second toes in the static footprints and at the heel in the dynamic footprints., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Chartered Society of Forensic Sciences. Published by Elsevier B.V. All rights reserved.)

Environmental trace evidence offers useful circumstantial intelligence to link persons and scenes of forensic interest. An increasing empirical research base is dedicated towards understanding the transfer and persistence dynamics of environmental indicators including pollen, soils, and diatoms, within a diverse range of experimental frameworks. This paper presents two discrete studies exploring transfer and persistence of soils and sediments on footwear and diatomaceous earth adhered to clothing in forensically pertinent scenarios. Variables including sediment type, foot position, clothing type, and body positioning were also explored throughout. Both experiments incorporated a field-based methodology during the sampling effort. Photographs were collected of an initial transfer sample and of a retained assemblage following hours, days, and up to one-week of wear, facilitating macroscopic assessment of trace evidence dynamics. All images were processed using accessible, open-source software before spatial analysis of evidence distribution within and temporal assessment (% retention) upon each evidential surface. The results highlighted consistent loss of transferred sediment from footwear with significantly greater retention of loamy clay soil than dune sand which was absent beyond 24 h of wear. Loss was not influenced by wearer gait but was more rapid from those areas of the shoe sole in direct contact with the ground. Diatomaceous earth was retrieved from all three clothing types tested after one week - significant losses of material occurred before 48 h with a consistent assemblage identified beyond this. Denim was significantly more effective than acrylic and fleece for diatomaceous earth retention and significantly more material was lost from clothing worn on the lower body. These findings highlight the value of using visual environmental markers and a macroscopic analytical approach during the investigation of environmental trace dynamics. The methodology offers a novel, non-destructive assessment of soil and diatom transfer and persistence, complementing more extensive laboratory-based examinations to ensure the development of a well-rounded research base within the forensic sciences., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Recently, digital forensics has become increasingly important as it is used by investigation agencies, corporate, and private sector. To supplement the limitations of evidence capacity and be recognized in court, it is essential to establish an environment that ensures the integrity of the entire process ranging from collecting and analyzing to submitting digital evidence to court. In this study, common elements were extracted by comparing and analyzing ISO/IEC 17025, 27001 standards and Interpol and Council of Europe (CoE) guidelines to derive the necessary components for building a digital forensic laboratory. Subsequently, based on 21 digital forensic experts in the field, Delphi survey and verifications were conducted in three rounds. As a result, 40 components from seven areas were derived. The research results are based on the establishment, operation, management, and authentication of a digital forensics laboratory suitable for the domestic environment, with added credibility through collection of the opinions of 21 experts in the field of digital forensics in Korea. This study can be referred to in establishing digital forensic laboratories in national, public, and private digital forensic organizations as well as for employing as competency measurement criteria in courts to evaluate the reliability of the analysis results., (© 2023 American Academy of Forensic Sciences.)

Analysis of evidence is a challenge. Crime scene materials are complex, diverse, sometimes of an unknown nature. Forensic science provides the most critical applications for their examination. Chemical tests, analytical methods, and techniques to process the evidence must be carefully selected by the forensic scientist. Ideally, it may be interpreted, analyzed, and judged in the original context of the crime scene. In this sense, hyperspectral imaging (HSI) has been employed as an analytical tool that maintains the integrity of the samples/objects for multiple and sequential analysis and for counter-proof exams. This paper is an overview of forensic science trends for the application of HSI techniques in the last ten years (2011-2021). The examination of documents was the main area of exploration, followed by bloodstain analysis aging process; trace analysis of explosives and gunshot residue. Chemometric tools were also addressed since they are crucial to obtain the most important information from the samples. There are great challenges in applying HSI in forensic science, but there have been clear technological and scientific advances, and a solid foundation has been built for the use of HSI in real-life cases., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Chartered Society of Forensic Sciences. Published by Elsevier B.V. All rights reserved.)

Diatoms (Bacillariophyta) are unicellular photosynthetic organisms commonly occurring in aquatic habitats on Earth. Their autecology makes them almost perfect indicators of environmental conditions and so have high potential for use in forensics. Both eurytopic and stenotopic species are important in site identification: eurytopic forms due to their abundance, and stenotopic forms due to their narrow range of tolerance to environmental conditions. Their presence can hence provide a clear insight into an ecosystem and its microhabitats. The diatoms are useful as significant indicator in diagnosing of drowning. However, to definitively link a corpse with the place where it was found and to indicate whether this was also the crime scene, it is essential that any comparative diatomaceous analyses are performed correctly. The following study presents selected cases in which a comparative diatomaceous analysis was performed. In all cases, the biological samples secured during the autopsy were compared with environmental samples collected from the site of the cadaver disclosure. Our findings show both the possibilities and limitations of using this method in forensics. These forensic investigations need close collaboration between coroners and diatomology experts. It is crucially important to reveal whether the place of corpse finding is the same as the place of drowning., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Forensic gait analysis is used to visually and quantitatively analyze information regarding the appearance and style of walking that can be presented as evidence in the court. The demand for analyzing CCTV pedestrian footage in video surveillance has been increasing. The dependence of the accuracy of semiautomatic silhouette-based analysis, often used in forensic science, on the differences in the viewing directions is a very challenging issue that is yet to be resolved for real case applications. Currently, the different viewing directions used in comparison footage significantly decrease the accuracy of same person analysis when using the silhouette-based method, often used in the Japanese forensic science domain. A calibration-based method was previously prosed to resolve this problem, but it requires performing an elaborate measurement procedure at the camera installation site for an accurate analysis. In this study, we propose a novel in-silico silhouette-based analysis method that significantly expands the number of viewing direction pre-set settings to 900 from the 24 used in the previous method. Several software tools have been developed to ensure that all the procedures can be executed on a computer. The experimental results confirm that the accuracy of the proposed method is comparable to that of the calibration-based method. Furthermore, the practical comparison results from actual consultation confirmed the effectiveness of the proposed method under existing viewing direction differences. We therefore anticipate that the proposed method will be beneficial for improving the analysis accuracy in real cases and therefore serve as a substitute of the previous method., (© 2023 American Academy of Forensic Sciences.)

Forensic entomology includes the analysis of organisms colonizing various parts of the body in order to determine the circumstances of an incident, mainly the time, place, and cause of death. The presence of insects and other arthropods on carcasses can be a source of knowledge for the judicial system. However, this type of research (on submerged bodies) is less published. The aim of our study was to analyse the qualitative and quantitative structure of macroinvertebrates colonizing potential evidence in an upland river. The experimental research involved an eight-week exposure to articles of clothing made of different materials: natural materials (bottom sediments with plants from a river), synthetic (socks), and cotton (t-shirts). Control samples of water after 2, 4, 6, and 8 weeks were taken from experiment locations in the River Bystrzyca with a tube apparatus and hand net. The results indicated that the abundance of organisms on a given substrate depended on the period of development of invertebrate macrofauna and the time of exposure of the substrates. The abundance of aquatic macrofauna on the exposed items increased in direct proportion to the duration of the experiment, which may indicate the adaptability of these organisms to new habitat conditions. Among the taxonomic groups used in forensic entomology, Diptera, Coleoptera, and Odonata were the most abundant. The remaining taxa (including Heteroptera), though not widely used in judicial proceedings, can also provide valuable information about the circumstances of an incident.

Recent advances in next-generation sequencing technologies (NGS) coupled with machine learning have demonstrated the potential of microbiome-based analyses in applied areas such as clinical diagnostics and forensic sciences. Particularly in forensics, microbial markers in biological stains left at a crime scene can provide valuable information for the reconstruction of crime scene cases, as they contain information on bodily origin, the time since deposition, and donor(s) of the stain. Importantly, microbiome-based analyses provide a complementary or an alternative approach to current methods when these are limited or not feasible. Despite the promising results from recent research, microbiome-based stain analyses are not yet employed in routine casework. In this review, we highlight the two main gaps that need to be addressed before we can successfully integrate microbiome-based analyses in applied areas with a special focus on forensic casework: one is a comprehensive assessment of the method's strengths and limitations, and the other is the establishment of a standard operating procedure. For the latter, we provide a roadmap highlighting key decision steps and offering laboratory and bioinformatic workflow recommendations, while also delineating those aspects that require further testing. Our goal is to ultimately facilitate the streamlining of microbiome-based analyses within the existing forensic framework to provide alternate lines of evidence, thereby improving the quality of investigations.

Field research within latent print comparison has remained sparse in the context of an otherwise growing body of literature examining the discipline. Studies examining how ACE-V procedures are implemented within active crime laboratories are especially lacking in light of research suggesting significant variability in examiner practices despite standardized ACE-V procedures. To date, no studies have examined a potentially important aspect of the Analysis phase: digital image editing. We provide information on the prevalence and types of latent print image editing within one laboratory (i.e., Houston Forensic Science Center), examine the potential effect of image editing on objective print quality and clarity (i.e., LQMetrics scores), and explore potential examiner differences in editing effectiveness. Results indicate that most latent prints are edited in some manner, and that image editing improves the quality and clarity of print images as defined by an objective quality metric, although examiners varied in their ability to improve the clarity of print images. Findings suggest that formal guidance or documentation of standard editing procedures would likely improve the reliability of examiner conclusions early in the latent print comparison process., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Chartered Society of Forensic Sciences. Published by Elsevier B.V. All rights reserved.)

Given the size and complexity of many digital forensic science device examinations, there is a need for practitioners to formally and strategically determine a course of conduct which allows them to undertake the most robust and efficient examination possible. This work outlines both the need for practitioners to have a digital evidence strategy (DES) when tackling any given examination scenario, how to construct one and the concerns which exist when no formal DES is in place. Approaches to DES development are examined and the context to which they should be deployed are analysed, with focus being on the use of DESs at the examination/processing stage of the investigative workflow. Finally, a 'DES skeleton' is offered to guide practitioners as they seek to create their own DES., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Chartered Society of Forensic Sciences. Published by Elsevier B.V. All rights reserved.)

The blood fingerprint enhancement is not so eye-catching as latent fingerprint development in forensic community, but it is indeed an important piece of evidence for personal identification, forensic analysis and even reconstruction of crime scenes. In over past ten years, novel reagents, advanced materials and emerging techniques have growingly participated in blood fingerprint enhancement, which not only leads to a higher level of developing sensitivity, selectivity and contrast, but also endows blood impressions with more forensic significance. This review summarizes recent advances in conventional chemical reagents targeting at heme, protein and amino acid as well as emerging enhancement techniques based on advanced materials, new equipment or methods. Some critical issues in forensic science are also discussed, including partial blood fingerprint enhancement, false positive of developing reagents, the compatibility of blood enhancement technique and DNA, fingerprint age determination, and so on. Finally, we have proposed several urgent problems to be solved and the prospects of some promising techniques were proposed in the field of blood fingerprint enhancement in future work.

Visualizing latent evidence at a crime scene has gained popularity in the field of forensic science during the past few years. Thus, this study designs and develops a one-click device for the rapid visualization and extraction of latent evidence through multimodal light source integration and light-guiding technology. Our device exhibits multispectral and angle timing functions for storing the captured evidence images. Furthermore, the geometric registration, feature extraction, feature optimization, and feature integration of the evidence images are processed by a backend system, and the images are then presented. Overall, this study enhances the standard and the technical content of evidence extraction and simplifies the evidence extraction process. In addition to the rapid handling of the scenes captured at a crime scene, the one-click device has other notable advantages, such as fast imaging, portability, being independent of the environmental conditions and the operator's technical capabilities, and zero pollution to ensure the repeatability of material evidence extraction. Compared with the original optical forensics equipment, the spectrum and angle of our system are more extensive., (© 2022. The Author(s).)

Document dating is an important and challenging task in the field of forensic science. With the development of analytical techniques, related methods are emerging for document dating studies. Desorption electrospray ionization-mass spectrometry (DESI-MS) is an ambient ionization-MS technique that has been applied in the forensic discrimination of inks. This study focused on utilizing DESI-MS to distinguish between ink entries of different ages. Blue ballpoint ink and black gel ink samples were artificially aged by heat and light exposure and then screened by DESI-MS in the positive ion mode, with a methanol-water-formic acid solution (90:10:0.1%, v/v/v) as the desorption solvent. As a result, the artificially aged ink samples were directly differentiated from the reference ones in the chemical images of age-dependent compounds such as basic dyes or polyethylene glycol derivatives. The amount of ethoxylated fatty amine surfactants in copy paper was also found to decrease after artificial aging. Moreover, this method enabled us to visualize the differences between the composition of naturally aged and fresh ink samples. It has been proven to cause no apparent destruction of the samples. The identification of chronological changes in document materials suggests that the DESI-MS method is a promising approach for relative document dating., (© 2022 American Academy of Forensic Sciences.)

The increasing volume, variety, velocity, distribution, structural intricacy, and complexity of use of digital evidence can make it difficult for practitioners to find and understand the most forensically useful information (Casey E. Digital evidence and computer crime: Forensic science, computers, and the Internet. Academic Press; 2011. p. 31; Pollitt M. The hermeneutics of the hard drive: Using narratology, natural language processing, and knowledge management to improve the effectiveness of the digital forensic process [PhD dissertation]. University of Central Florida; 2011). Digital forensic practitioners currently search for information and solutions in an ad hoc manner, leading to results that are unstructured, unverified, and sometimes incomplete. As a result, certain digital evidence is being missed or misinterpreted. To mitigate risks of knowledge gaps, there is a pressing need for a systematic mechanism that practitioners can use to codify and combine their collective knowledge. This work presents the design and development of a solution that catalogs crowdsourced knowledge of digital forensic artifacts in a well-structured, easily searchable form to support efficient and automated extraction of pertinent information, improving availability and reliability of interpretation of artifacts (general acceptance). Technical implementation and artifact curation are discussed with illustrative examples and recommendations for future work., (Published 2022. This article is a U.S. Government work and is in the public domain in the USA. Journal of Forensic Sciences published by Wiley Periodicals LLC on behalf of American Academy of Forensic Sciences.)

Ground Truth Data is data that comes from a known source, where the truth about the data is known and not inferred (FSR, 2021). The Forensic Science Regulator requires forensic units (that carry out certain forensic processes) to undertake tests against ground truth data for the purposes of quality assurance/quality control processes such as accreditation. However, the data collected must form a meaningful dataset that will enable relevant tests to be performed that inform the end-user. This technical note discusses how a forensic unit in the UK planned and developed a method for collecting ground truth data for footwear mark evidence. It discusses the materials and variables that were considered when developing the method and the evidence-based, decision-making processes that enabled its creation. Recommendations and considerations are provided to assist other forensic units collect data relevant to their jurisdiction. Whilst the method is not prescriptive, if it is used as a guide, it may facilitate the development of a large and relevant national dataset., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Chartered Society of Forensic Sciences. Published by Elsevier B.V. All rights reserved.)

Forensic handwriting examination involves the comparison of writing samples by forensic document examiners (FDEs) to determine whether or not they were written by the same person. Here we report the results of a large-scale study conducted to assess the accuracy and reliability of handwriting comparison conclusions. Eighty-six practicing FDEs each conducted up to 100 handwriting comparisons, resulting in 7,196 conclusions on 180 distinct comparison sets, using a five-level conclusion scale. Erroneous "written by" conclusions (false positives) were reached in 3.1% of the nonmated comparisons, while 1.1% of the mated comparisons yielded erroneous "not written by" conclusions (false negatives). False positive rates were markedly higher for nonmated samples written by twins (8.7%) compared to nontwins (2.5%). Notable associations between training and performance were observed: FDEs with less than 2 y of formal training generally had higher error rates, but they also had higher true positive and true negative rates because they tended to provide more definitive conclusions; FDEs with at least 2 y of formal training were less likely to make definitive conclusions, but those definitive conclusions they made were more likely to be correct (higher positive predictive and negative predictive values). We did not observe any association between writing style (cursive vs. printing) and rates of errors or incorrect conclusions. This report also provides details on the repeatability and reproducibility of conclusions, and reports how conclusions are affected by the quantity of writing and the similarity of content.

A critical concern with crime scene documentation is the accuracy with which a crime scene can be reconstructed. Here, we discuss the accuracy of eight documentation methods as a function of measurement distance between reference ground targets in an outdoor scene. The relative accuracy of each documentation method was assessed with respect to a widely accepted and well-established standard method for land surveying, Total Station, from which measurements served as "ground truth" or reference data. For the majority of methods, the actual relative difference between measurements when compared to Total Station was small (less than a quarter of an inch). Measurements from FARO LiDAR agreed the most with to those of Total Station, while drone without the use of ground control points (GCPs) agreed the least. GCPs or a reference scale were also found to be important in mitigating increasing imprecision with increasing distance when measuring between two targets ~9-85 ft apart via drone and orthomosaic methods. Additionally, there were no statistical differences in the use of 2D (horizontal) or 3D (slope) measurement configurations for the Total Station. Overall, linear regression of difference plots did not reveal meaningful correlation between increasing distance measured and the error of a method when compared to Total Station. As more measurement methods become available, and the need for training and validating new tools become a necessity, these results point to the importance of establishing a ground truth or known distance range on which crime scene measurement methods can be validated., (Published 2022. This article is a U.S. Government work and is in the public domain in the USA.)

Individual identification is one of the research hotspots in the practice of forensic science, and the judgment is usually built on the comparison of the unique biological characteristics of the individual, such as fingerprints, iris and DNA. With the dramatic increase in the number of cases related to video image investigations, there is an increasing need for the technology to identify individuals based on the macroscopic comparison of facial appearance biometrics. At present, with the introduction of computer three-dimensional (3D) modeling and 3D superimposition comparison technology, considerable progress has been made in individual identification methods based on macroscopic comparison of facial appearance biometrics. This paper reviews individual facial appearance biometric methods based on macroscopical comparison, comprehensively analyzes the advantages and limitations of different methods, and puts forward recommendations and prospects for subsequent research.

Surface analysis techniques have rapidly evolved in the last decade. Some of these are already routinely used in forensics, such as for the detection of gunshot residue or for glass analysis. Some surface analysis approaches are attractive for their portability to the crime scene. Others can be very helpful in forensic laboratories owing to their high spatial resolution, analyte coverage, speed, and specificity. Despite this, many proposed applications of the techniques have not yet led to operational deployment. Here, we explore the application of these techniques to the most important traces commonly found in forensic casework. We highlight where there is potential to add value and outline the progress that is needed to achieve operational deployment. We consider within the scope of this review surface mass spectrometry, surface spectroscopy, and surface X-ray spectrometry. We show how these tools show great promise for the analysis of fingerprints, hair, drugs, explosives, and microtraces.

Introduction and Objectives: The human external ear is a morphologically perplexing structure with wide variations in its features across individuals. Advances in forensic science have established the use of morphological features and prints of different parts of the human body in establishing the identity of an individual. The present study aims to establish such an identification process by examining the use of comparative analysis of ear prints and ear photographs., Methodology: The study examined the morphological features of ear prints of 140 participants (71 males and 69 females) ranging in age from 18 to 25 years (with a mean age of 20.22 ± 1.77 years), recruited from a North Indian population. Prints of both the ears were recorded, using the inking method. The morphological features of the ear prints were compared with the photographs of the ear for their uniqueness and individualization using likelihood ratios., Results: The degree ofantihelix curvature demonstrated the highest correlation (98.57%) between the morphological features visible on the ear and the expressions of these features in an ear print. The impression of earlobes, Darwin tubercle, crus of the helix, and shape of the external ear showed accuracy between 70% and 90%, whereas correlations below 70% accuracy were recorded for the other features. The likelihood ratios for 12 parameters were reported to be more than 10x, representing that the identification made using these parameters was > 10 times more likely to be true identification as compared to false positives. Moreover, 5 parameters were found to show high sensitivity, with values greater than 3 standard deviations (0.997), with an additional 9 parameters found to have good sensitivity at values greater than 2 standard deviations (<0.95). Similarly, 2 parameters were found to show high specificity, with values greater than 3 standard deviations (0.997), with an additional 18 parameters found to have good sensitivity at values greater than 2 standard deviations (<0.95)., Conclusions: Elevated features of the ear, such as helix, antihelix, antitragus, tragus, and inter-tragic notch; particularly the degree of antihelix curvature, are more easily impressed and identifiable on an ear print. The results support that ear prints may provide important clues to assist in establishing the identity of an individual if several morphological features are evaluated concurrently., Competing Interests: The authors report no conflicts of interest., (Copyright © 2021 by Mutaz B. Habal, MD.)

Forensic facial image comparison lacks a methodological standardization and empirical validation. We aim to address this problem by assessing the potential of machine learning to support the human expert in the courtroom. To yield valid evidence in court, decision making systems for facial image comparison should not only be accurate, they should also provide a calibrated confidence measure. This confidence is best conveyed using a score-based likelihood ratio. In this study we compare the performance of different calibrations for such scores. The score, either a distance or a similarity, is converted to a likelihood ratio using three types of calibration following similar techniques as applied in forensic fields such as speaker comparison and DNA matching, but which have not yet been tested in facial image comparison. The calibration types tested are: naive, quality score based on typicality, and feature-based. As transparency is essential in forensics, we focus on state-of-the-art open software and study their power compared to a state-of-the-art commercial system. With the European Network of Forensic Science Institutes (ENFSI) Proficiency tests as benchmark, calibration results on three public databases namely Labeled Faces in the Wild, SC Face and ForenFace show that both quality score and feature based calibration outperform naive calibration. Overall, the commercial system outperforms open software when evaluating these Likelihood Ratios. In general, we conclude that calibration implemented before likelihood ratio estimation is recommended. Furthermore, in terms of performance the commercial system is preferred over open software. As open software is more transparent, more research on open software is urged for., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Crime scene investigation serves as an important source of information for law enforcement officers when running a case. Effective leads are uncovered based on the knowledge, technique, and experience of the crime scene investigator. This study utilized eye-tracking technology to understand different decision makers'visual behaviors and decision-making process when studying a crime scene picture. A total of 42 college students majored in crime investigation were recruited as the subjects of the study. Eye movement data were collected and analyzed using t-test and ANOVA on eye-tracking measures of areas of interest (AOI) for visual attention distributions as well as lag sequence analysis (LSA), heat map, and scan path observations for visual transfer patterns. The results showed that participants who made the correct decision were faster in identifying the key target. Regardless of whether they made the correct decision or not, their visual attention was fixated on the answer option of their choosing. Additionally, the subjects' attention must be shifted to the key target in order to make the correct decision. This study successfully applied the eye-tracking method to explore the investigation and decision-making process of inspecting a crime scene picture. Future studies can apply the eye-tracking method to deeply understand the process of information processing in investigating criminal scene pictures and provide more suggestions for professional training curriculum and instruction., (© 2022 American Academy of Forensic Sciences.)

Forensic entomology is the branch of forensic science that is related to using arthropod specimens found in legal issues. Fly maggots are one of crucial pieces of evidence that can be used for estimating post-mortem intervals worldwide. However, the species-level identification of fly maggots is difficult, time consuming, and requires specialized taxonomic training. In this work, a novel method for the identification of different forensically-important fly species is proposed using convolutional neural networks (CNNs). The data used for the experiment were obtained from a digital camera connected to a compound microscope. We compared the performance of four widely used models that vary in complexity of architecture to evaluate tradeoffs in accuracy and speed for species classification including ResNet-101, Densenet161, Vgg19&#95;bn, and AlexNet. In the validation step, all of the studied models provided 100% accuracy for identifying maggots of 4 species including Chrysomya megacephala (Diptera: Calliphoridae), Chrysomya (Achoetandrus) rufifacies (Diptera: Calliphoridae), Lucilia cuprina (Diptera: Calliphoridae), and Musca domestica (Diptera: Muscidae) based on images of posterior spiracles. However, AlexNet showed the fastest speed to process the identification model and presented a good balance between performance and speed. Therefore, the AlexNet model was selected for the testing step. The results of the confusion matrix of AlexNet showed that misclassification was found between C. megacephala and C. (Achoetandrus) rufifacies as well as between C. megacephala and L. cuprina. No misclassification was found for M. domestica. In addition, we created a web-application platform called thefly.ai to help users identify species of fly maggots in their own images using our classification model. The results from this study can be applied to identify further species by using other types of images. This model can also be used in the development of identification features in mobile applications. This study is a crucial step for integrating information from biology and AI-technology to develop a novel platform for use in forensic investigation., (© 2022. The Author(s).)