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Hyperspectral imaging (HSI) has become a crucial innovation in forensic science, particularly for analysing bodily fluids. This advanced technology captures both spectral and spatial data across a wide spectrum of wavelengths, offering comprehensive insights into the composition and distribution of bodily fluids found at crime scenes. In this review, we delve into the forensic applications of HSI, emphasizing its role in detecting, identifying, and distinguishing various bodily fluids such as blood, saliva, urine, vaginal fluid, semen, and menstrual blood. We examine the benefits of HSI compared to traditional methods, noting its non-destructive approach, high sensitivity, and capability to differentiate fluids even in complex mixtures. Additionally, we discuss recent advancements in HSI technology and their potential to enhance forensic investigations. This review highlights the importance of HSI as a valuable tool in forensic science, opening new pathways for improving the accuracy and efficiency of crime scene analyses. • Hyperspectral imaging enhances non-destructive detection of fluids at crime scenes. • Ability to precisely identify and differentiate complex fluid mixtures. • Various detection methods for blood, saliva, urine, vaginal fluid, & semen. • Compares hyperspectral imaging with traditional forensic methods. • Adaptability provides advantages over conventional techniques. [ABSTRACT FROM AUTHOR]

Given that microbiological analysis can be an alternative method that overcomes the shortcomings of traditional forensic technology, and skin samples may be the most common source of cases, the analysis of skin microbiome was investigated in this study. High-throughput sequencing targeting the V3-V4 region of 16S rRNA gene was performed to reveal the skin microbiome of healthy individuals in Guangdong Han. The bacterial diversity of the palm, navel, groin and plantar of the same individual was analyzed. The overall classification based on 16S rRNA gene amplicons revealed that the microbial composition of skin samples from different anatomical parts was different, and the dominant bacterial genus of the navel, plantar, groin and palm skin were dominated by Cutibacterium , Staphylococcus , Corynebacterium and Staphylococcus , respectively. PCoA analysis showed that the skin at these four anatomical locations could only be grouped into three clusters. A predictive model based on random forest algorithm showed the potential to accurately distinguish these four anatomical locations, which indicated that specific bacteria with low abundance were the key taxa. In addition, the skin microbiome in this study is significantly different from the dominant microbiome in saliva and vaginal secretions identified in our previous study, and can be distinguished from these two tissue fluids. In conclusion, the present findings on the community and microbial structure details of the human skin may reveal its potential application value in assessing the location of skin samples and the type of body fluids in forensic medicine. • In this study, four skin sites showed differences in the abundance of dominant flora and specific low-abundance bacteria. • The random forest model showed high accuracy in distinguishing skin locations of four different anatomical structures. • Microbial composition of skin, saliva and vaginal secretions varies widely for inferring forensic tissue fluid types. [ABSTRACT FROM AUTHOR]

Criminal investigations, particularly sexual assaults, frequently require the identification of body fluid type in addition to body fluid donor to provide context. In most cases this can be achieved by conventional methods, however, in certain scenarios, alternative molecular methods are required. An example of this is the detection of menstrual fluid and vaginal material, which are not able to be identified using conventional techniques. Endpoint reverse-transcription PCR (RT-PCR) is currently used for this purpose to amplify body fluid specific messenger RNA (mRNA) transcripts in forensic casework. Real-time quantitative reverse-transcription PCR (RT-qPCR) is a similar method but utilises fluorescent markers to generate quantitative results in the form of threshold cycle (C q) values. Despite the uncertainty surrounding body fluid identification, most interpretation guidelines utilise categorical statements. Probabilistic modelling is more realistic as it reflects biological variation as well as the known performance of the method. This research describes the application of various machine learning models to single-source mRNA profiles obtained by RT-qPCR and assesses their performance. Multinomial logistic regression (MLR), Naïve Bayes (NB), and linear discriminant analysis (LDA) were used to discriminate between the following body fluid categories: saliva, circulatory blood, menstrual fluid, vaginal material, and semen. We identified that the performance of MLR was somewhat improved when the quantitative dataset of the original C q values was used (overall accuracy of approximately 0.95) rather than presence/absence coded data (overall accuracy of approximately 0.94). This indicates that the quantitative information obtained by RT-qPCR amplification is useful in assigning body fluid class. Of the three classification methods, MLR performed the best. When we utilised receiver operating characteristic curves to observe performance by body fluid class, it was clear that all methods found difficulty in classifying menstrual blood samples. Future work will involve the modelling of body fluid mixtures, which are common in samples analysed as part of sexual assault investigations. [ABSTRACT FROM AUTHOR]

The identification of body fluids at a crime scene is an important aspect of forensic casework analysis, being a source for investigative leads and contributing to case evidence. Yet, current methods for the forensic identification of body fluids suffer from several limitations, ranging from poor sensitivity and specificity, to sample destruction and interference with subsequent DNA analysis. Moreover, current identification assays target only one body fluid at the time. Besides being inefficient in terms of time, money and sample consumption, poor identification methods can also negatively influence the outcome of a (court) case. In this study, eleven potential protein biomarkers and antibodies were selected and assessed on their suitability for serving as identification markers, as a first step towards the development of a new multiplex protein-based body fluid identification assay relying on antigen-antibody interactions. Performing antibody-based dot blot assays, the specificity of the biomarkers for their target body fluids was evaluated, and biomarker detection was studied in diluted, mixed, aged and simulated casework samples. Hereby, nine out of eleven markers were identified as promising biomarkers to identify blood, semen, saliva, urine and sweat. With the identification of these targets and detection antibodies, a major step forward has been taken towards the development of a highly sensitive and specific, fast and non-labour-intensive protein-based body fluid identification assay, suitable for on-site analysis and able to test for multiple body fluids in a single reaction. [ABSTRACT FROM AUTHOR]

The Phadebas® Forensic Press Test is routinely used for the detection of saliva. However, assessment of the use of Phadebas® paper for this purpose has not been studied extensively. The suitability of Phadebas® paper as a presumptive screening tool for saliva on forensic exhibits, was investigated by analysing the following: (1) sensitivity, (2) specificity, (3) effects of temperature on sensitivity and specificity, (4) detection of saliva in mixed body fluid samples, and (5) influence of substrate porosity. The results of this study demonstrated that Phadebas® paper is more sensitive to α-amylase activity and less specific for saliva than previously reported. The use of an examination temperature of 37°C had no effect on sensitivity, but increased the incidence of cross-reactivity with other forensically relevant body fluid stains. Blood, urine and vaginal secretions can inhibit the detection of α-amylase activity with Phadebas® paper in mixed stains of saliva and body fluid. Substrate porosity is a weak predictor for the time taken for a saliva stain to achieve a strong positive result on Phadebas® paper. Overall, this study demonstrated that the Phadebas® Forensic Press Test has limitations as a presumptive test for the accurate identification of saliva. [ABSTRACT FROM AUTHOR]

Field based forensic tests commonly provide information on the presence and identity of biological stains and can also support the identification of species. Such information can support downstream processing of forensic samples and generate rapid intelligence. These approaches have traditionally used chemical and immunological techniques to elicit the result but some are known to suffer from a lack of specificity and sensitivity. The last 10 years has seen the development of field-based genetic profiling systems, with specific focus on moving the mainstay of forensic genetic analysis, namely STR profiling, out of the laboratory and into the hands of the non-laboratory user. In doing so it is now possible for enforcement officers to generate a crime scene DNA profile which can then be matched to a reference or database profile. The introduction of these novel genetic platforms also allows for further development of new molecular assays aimed at answering the more traditional questions relating to body fluid identity and species detection. The current drive for field-based molecular tools is in response to the needs of the criminal justice system and enforcement agencies, and promises a step-change in how forensic evidence is processed. However, the adoption of such systems by the law enforcement community does not represent a new strategy in the way forensic science has integrated previous novel approaches. Nor do they automatically represent a threat to the quality control and assurance practices that are central to the field. This review examines the historical need and subsequent research and developmental breakthroughs in field-based forensic analysis over the past two decades with particular focus on genetic methods Emerging technologies from a range of scientific fields that have potential applications in forensic analysis at the crime scene are identified and associated issues that arise from the shift from laboratory into operational field use are discussed. [ABSTRACT FROM AUTHOR]

The identification of vaginal fluids is crucial in forensic science. Several molecular protocols based on PCR amplification of mfDNA (microflora DNA) specific for vaginal bacteria are now available. Unfortunately mfDNA extraction and PCR reactions require manual optimization of several steps. The aim of present study was the verification of a partial automatization of vaginal fluids identification through two instruments widely diffused in forensic laboratories: EZ1 Advanced robot and Rotor Gene Q 5Plex HRM. Moreover, taking advantage of 5-plex thermocycler technology, the ForFluid kit performances were improved by expanding the mfDNA characterization panel with a new bacterial target for vaginal fluids and with an internal positive control (IPC) to monitor PCR inhibition. Results underlined the feasibility of a semi-automated extraction of mfDNA using a BioRobot and demonstrated the analytical improvements of the kit. [ABSTRACT FROM AUTHOR]

Introduction: Oral fluid testing is widely used for detecting drug exposure, but data describing methadone and metabolites in oral fluid during pharmacotherapy for opioid-dependence are relatively limited.Methods: 414 oral fluid specimens from 16 opioid-dependent pregnant women receiving daily methadone were analyzed for methadone, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), and methadol by liquid chromatography-mass spectrometry.Results: All oral fluid specimens contained methadone greater than 1 ng/mL; 88% were positive for EDDP and 12% for methadol. Over 95% of oral fluid specimens exceeded the 20 ng/mL methadone cutoff set by the European Driving Under the Influence of Drugs, Alcohol and Medicines (DRUID) study. Methadone and EDDP oral fluid concentrations were highly variable within and between participants, did not predict methadone dose, but were negatively correlated with pH.Conclusion: Methadone was readily identified in oral fluid at concentrations greater than 20 ng/mL following daily 30-110 mg/day methadone pharmacotherapy. As no specimens contained only EDDP or methadol, there was no advantage to including these analytes for identification of methadone exposure. As nearly all oral fluid specimens from methadone-maintained patients exceeded the DRUID guideline, the 20 ng/mL cutoff appears to be sensitive enough to detect daily methadone exposure; however, additional indicators of behavioral and/or motor impairment would be necessary to provide evidence of driving impairment. [ABSTRACT FROM AUTHOR]

Abstract: This study has been extended to compare directly the accuracy of determining death interval from biochemical parameter of synovial fluid and vitreous humour. In 154 medicolegal cases with known time of death both fluids were taken and analyzed for sodium, potassium, chloride, calcium, creatinine, glucose, and urea. Both fluids showed comparable values. The study indicates that level of potassium and glucose levels in synovial fluid for determining death interval are atleast as accurate as if not more accurate than potassium and glucose levels in vitreous humour. Nevertheless determination of level of potassium in synovial fluid and vitreous humour can afford the most accurate method of determining death interval. [ABSTRACT FROM AUTHOR]

Abstract: Aim: To study several methods for estimating the prevalence of high blood concentrations of tetrahydrocannabinol and amphetamine in a population of drug users by analysing oral fluid (saliva). Methods: Five methods were compared, including simple calculation procedures dividing the drug concentrations in oral fluid by average or median oral fluid/blood (OF/B) drug concentration ratios or linear regression coefficients, and more complex Monte Carlo simulations. Populations of 311 cannabis users and 197 amphetamine users from the Rosita-2 Project were studied. Results: The results of a feasibility study suggested that the Monte Carlo simulations might give better accuracies than simple calculations if good data on OF/B ratios is available. If using only 20 randomly selected OF/B ratios, a Monte Carlo simulation gave the best accuracy but not the best precision. Dividing by the OF/B regression coefficient gave acceptable accuracy and precision, and was therefore the best method. None of the methods gave acceptable accuracy if the prevalence of high blood drug concentrations was less than 15%. Conclusion: Dividing the drug concentration in oral fluid by the OF/B regression coefficient gave an acceptable estimation of high blood drug concentrations in a population, and may therefore give valuable additional information on possible drug impairment, e.g. in roadside surveys of drugs and driving. If good data on the distribution of OF/B ratios are available, a Monte Carlo simulation may give better accuracy. [Copyright &y& Elsevier]

Abstract: We evaluated the performance of real-time RT-PCR and ELISA assays for detection of dermcidin (DCD) in sweat and body-fluid stains. DCD, a small antibiotic peptide secreted into human sweat, was detected by real-time RT-PCR in 7-day-old stains containing as small as 10μL of sweat, and the assay showed high specificity when testing 7-day-old stains containing 30μL of other body-fluid. ELISA using anti-human dermcidin mouse monoclonal antibody detected DCD sweat diluted up to approximately 10,000-fold and could specifically detect DCD in 10μL of body-fluid stains. The performance of the two assays was tested during winter on samples that simulated forensic case samples: an undershirt and a sock worn for 20h, a handkerchief used to wipe the brow several times within 12h, a cap and a cotton glove worn for 4h, and a white robe worn at intervals for 2 years. The result showed that the former assay detected DCD in all sites of the undershirt examined (armpit, back, and breast), and the latter gave a relatively high OD value in the armpit among the three sites. For the socks, although the latter assay gave very high OD values in both the center and toe of the foot sole, the former could not detect DCD in both of them. These results indicate that highly damp conditions, such as inside a shoe, might promote the degradation of mRNA in samples such as socks. In the other case samples, sweat was adequately detected by both assays. This study is the first demonstration of the use of real-time RT-PCR to sensitively identify sweat among body-fluid stains, and it confirmed that dermcidin was an excellent marker for sweat identification. In addition, the usefulness of ELISA was also verified. Positive sweat identification using these assays is expected to assist forensic practice. [Copyright &y& Elsevier]

Abstract: A great potential of Raman spectroscopy for non-destructive, confirmatory identification of body fluids at the crime scene has been reported recently (Virkler and Lednev, Forensic Sci. Int. 2008 ). However, that analysis was carried out on only one sample of each body fluid and did not take into account any variations that might occur between different donors of the same fluid. This paper reports on the role of heterogeneity within a sample as well as among multiple donors for human semen. Near-infrared (NIR) Raman spectroscopy was used to measure spectra of pure dried human semen samples from multiple donors in a controlled laboratory environment. The major chemical components that contributed to the Raman spectrum of semen were determined and used to tentatively identify the principal spectral components. The issue of potential spectral variations that could arise between different donors of semen was also addressed. Advanced statistical analysis of spectra obtained from multiple spots on dry samples showed that dry semen is heterogeneous and its Raman spectra could be presented as a linear combination of a fluorescent background and three spectral components. The relative contribution of each of the three components varies with donor, so no single spectrum could effectively represent an experimental Raman spectrum of dry semen in a quantitative way. The combination of the three spectral components could be considered to be a spectroscopic signature for semen. This proof-of-concept approach shows the potential for Raman spectroscopy to identify an unknown substance to be semen during forensic analysis. [Copyright &y& Elsevier]

Abstract: Body fluids and tissues in eight doxepin (Dox)-related deaths were investigated in order to prove whether the individual concentration of Dox, the concentration sum of parent drug and its active metabolite N-desmethyldoxepin (NDox) or the concentration ratio Dox/Ndox valuably contribute to making a cause of death determination. Individual case histories were shortly described. Dox and NDox concentrations were determined by LC–MS/MS. Dox concentration measured from two cases was well within a concentration range considered therapeutic, whereas subtherapeutic dosing may have occurred in another two cases. There were two cases of fatal Dox ingestion, as well as a case of high dosage and advanced putrefaction, respectively. The liver concentration sum may be more useful if a fatal ingestion cannot be clearly separated from a person''s medication usage. High concentrations could be observed in lung tissue, and combined concentrations of Dox and NDox may also be helpful in making a cause of death determination. There was a trend to a higher concentration sum in the brain with increasing combined levels in blood. Overall, the sum of the absolute figures allows a more accurate interpretation in Dox-related deaths as compared to the molar concentration ratio which may be helpful in acute ingestion. Determination of the N-desmethyl metabolite along with its parent is recommended and analysis should include more than a single specimen. [Copyright &y& Elsevier]

Abstract: Body fluid traces recovered at crime scenes are among the most important types of evidence to forensic investigators. They contain valuable DNA evidence which can identify a suspect or victim as well as exonerate an innocent individual. The first step of identifying a particular body fluid is highly important since the nature of the fluid is itself very informative to the investigation, and the destructive nature of a screening test must be considered when only a small amount of material is available. The ability to characterize an unknown stain at the scene of the crime without having to wait for results from a laboratory is another very critical step in the development of forensic body fluid analysis. Driven by the importance for forensic applications, body fluid identification methods have been extensively developed in recent years. The systematic analysis of these new developments is vital for forensic investigators to be continuously educated on possible superior techniques. Significant advances in laser technology and the development of novel light detectors have dramatically improved spectroscopic methods for molecular characterization over the last decade. The application of this novel biospectroscopy for forensic purposes opens new and exciting opportunities for the development of on-field, non-destructive, confirmatory methods for body fluid identification at a crime scene. In addition, the biospectroscopy methods are universally applicable to all body fluids unlike the majority of current techniques which are valid for individual fluids only. This article analyzes the current methods being used to identify body fluid stains including blood, semen, saliva, vaginal fluid, urine, and sweat, and also focuses on new techniques that have been developed in the last 5–6 years. In addition, the potential of new biospectroscopic techniques based on Raman and fluorescence spectroscopy is evaluated for rapid, confirmatory, non-destructive identification of a body fluid at a crime scene. [Copyright &y& Elsevier]

Abstract: Many plants contain toxic alkaloids which may be dangerous to humans. Despite the large number of poisonous plants, cases of fatal plant poisonings are relatively rare. The frequencies of poisonings and the plants involved are often regionally specific. Plant poisonings can be aggregated into three categories: unintended ingestions, intended ingestions, and poisoning due to abuse of plant material. Unintended ingestions often occur in children or from a mix-up of plants and mushrooms in adults. Intended ingestions are common in homicides and suicides. Increasingly common is the abuse of plants for hallucinogenic reasons. Toxicological analysis of such alkaloids may help in diagnosis of poisoning or abuse cases. This review describes the toxic alkaloids aconitine, atropine, coniine, colchicine, cytisine, dimethyltryptamine, harmine, harmaline, ibogaine, kawain, mescaline, scopolamine, and taxine, which are often involved in fatal and non-fatal poisonings. The paper summarizes the symptoms of the intoxications and reviews the methods of detection of their toxic constituents in biological fluids. [Copyright &y& Elsevier]

Abstract: The detection of diabetic coma postmortem requires accurate biochemical analysis. Due to continuous consumption of glucose by surviving cells postmortem, blood glucose levels decrease rapidly. Therefore, vitreous fluid has been used as a substitute in forensic practice, since it has a very low cell count. It has been repeatedly reported that the sum value of vitreous glucose and lactate should be used to estimate the original antemortem blood glucose level, based on the assumption that pre-existing glucose is gradually converted to lactate under anaerobic conditions during agonal phase and the early postmortem period. In this study, we applied a strategy including consistent sampling of vitreous fluid from the centre of both eyes of deceased subjects as soon as possible after arrival at the morgue, and immediate bedside analysis using a blood gas instrument. In total, 3076 cases were included during 2004–2006. We found that, after an initial drop of vitreous glucose during the very early postmortem period, the levels stayed stable for appreciable time postmortem. Analysis of a second sample collected at autopsy 1–3 days later gave similar results (R 2 =0.90). In contrast, the vitreous lactate levels showed a steady increase. This implies that the sum value of glucose and lactate increases with postmortem time, as reflected by vitreous potassium level. In fact, a statistically significant difference in the sum value was seen between subjects with potassium below 10mmol/L (n =1086) and above 20mmol/L (n =531), p <.001. In addition, in this large material, we did not identify a single case with circumstantial indication of hyperglycemia that only showed high vitreous lactate. We therefore suggest that vitreous glucose alone should be used to diagnose hyperglycemia postmortem and that the limit of 10mmol/L should have a good specificity for diabetic coma, which theoretically would equal an original blood glucose value of about 26mmol/L. As to the methodology, we found that sonication, centrifugation and addition of fluoride to the samples are unnecessary procedures when using a blood gas instrument. The strategy resulted in a doubling of the number of diabetic coma identified at our department compared to preceding period when analysis only was performed on selected cases. [Copyright &y& Elsevier]

Body fluid analysis has played a crucial role in ascertaining various characteristics and has greatly aided in reconstructing events during crime scene investigation. It is often presumed that crimes that involve violence and mental disturbances such as murder or sexual assault provide good sources of body fluids such as blood, saliva, semen, vaginal secretions, urine and tears. Tears are secreted in response to any emotional or stressful situations and may be found deposited on surfaces such as bedding, tissue paper or cloth. In the absence of the commonly noted body fluids such as blood or saliva, tears can play an important role that can lead to personal identification by examining the biochemistry and molecular aspects to obtain a full DNA profile. Additionally, identification of an individual may be done by carefully observing certain unique eye characteristics such as heterochromia which is highly individualistic. Characteristics of eyewear such as spectacles and contact lenses have unique properties and prescription criteria for correcting an individual's eyesight that can provide vital clues in understanding the visual ability of an individual. In crime scene investigation, the presence or absence of eyewear provides immense evidentiary value that has greatly aided in solving cases such as Janet Abaroa's Murder. This paper provides a systematic review of the possibility of using tears and eyewear for the purpose of forensic investigation and to statistically support the inferences with prescription databases which may be initiated across different populations. Forensic Optometry is yet to get streamlined along with the routinely followed investigative techniques and scientifically explored although no standard protocols exist to analyse eyewear. The use of behavioural optometry is gaining attention in the context of driving laws of different countries and is a simple but powerful indicator of abnormal behaviour. It is speculated that the last seen image referred to as an 'Optogram' of an individual may be captured in the retina since our eyes functions like a camera. Although this claim is considerably unexplored, it is quite possible that the last seen image of a criminal, objects or a place may be noted that can positively help in linking individuals at the scene of crime or identify the primary crime location. In this review, the potential for new insights into the analysis of tears, eye and eyewear characteristics have been explored. [ABSTRACT FROM AUTHOR]

Approximately during the 30th week of pregnancy, a woman gave birth to a still-born child in a hospital. After first citing an extraneous cause for the premature still-birth, the woman later admitted to having self-induced the abortion by injecting the antiseptic Rivanol® (active agent: ethacridine lactate) through her abdominal wall into the amniotic cavity. The investigating authorities ordered an autopsy of the fetus along with additional toxicological investigations. To the naked eye, no obvious cause of death was apparent. The main autopsy findings were four skin defects (puncture/stabbing wounds) on the ball of the fetus's left thumb, with slight bleeding around the punctures and into the underlying fatty tissue, and a yellowish discoloration of the fetus's body surface, especially of the umbilical cord and fingernails. On basis of the results, the child would have been viable. Femoral vein blood and urine from the fetus were analyzed for ethacridine, as were an amniotic fluid sample and maternal blood and urine samples, which had been collected as evidence. The concentration of ethacridine in the amniotic fluid was 16mg/l. In the postmortem fetal blood and urine samples, the concentrations were 0.36mg/l and 0.34mg/l, respectively, while concentrations of 0.091mg/l and 0.42mg/l, respectively, were found in the serum and urine samples from the mother. In many countries, foremost in China, ethacridine lactate, to which both mother and child are exposed, is widely used as safe abortion method. Although the ethacridine concentrations found in blood and urine samples of the mother in our case are consistent with published values, we believe to be the first to report postmortem ethacridine concentrations in a fetus. While exposure to ethacridine is not toxicologically relevant for the mother, it is fatal for the fetus because it causes the placental decidua capsularis to separate from the decidua parietalis or decidua placentalis, respectively. Prostaglandins that are then produced induce labor. In medicolegal contexts, the proof for an abortion through the administration of ethacridine lactate lies in the typical yellow discoloration of the fetus in conjunction with the toxicological demonstration of the substance in fetal body fluids, and if possible also in maternal body fluids. [ABSTRACT FROM AUTHOR]