Persistence of transferred fragrance on fabrics for forensic reconstruction applications

It has recently been established that volatile organic compounds (VOCs) successfully transfer between clothing even with a short contact of 10 s, highlighting the potential to use VOCs in forensic reconstruction scenarios, such as sexual assault cases. The mid and low volatility compounds transferred in greater amounts than high volatility compounds. This study presents empirical data addressing the persistence of transferred VOCs on clothing for the first time. A series of experiments were carried out to determine the persistence of VOCs on clothing for time periods of up 4 weeks, on natural and synthetic fibres, and at three different environmental temperatures. The data indicate that the highest VOC amounts are generally obtained for shorter persistence times of up to 1 d. Whilst high volatility compounds were not recovered in sufficient amounts to allow quantification, the four other transferred VOCs were successfully quantified for persistence times of up to 4 weeks. The persistence for mid-volatility compounds follows decay curve trends in line with those previously obtained for fibres, glass and pollen. When comparing the persistence of VOCs on a natural and a synthetic fibre, for a persistence time of 1 h, the transferred VOCs were retained on a natural fibre in higher amounts than on a synthetic fibre. However, for longer persistence times the concentration of VOCs was similar between the two fabrics. Lastly, lower environmental temperatures resulted in higher recoveries for most VOCs, especially for short persistence times. These findings demonstrate that optimal recovery of VOCs from clothing occurs when the fabric is kept at cooler temperatures and analysed soon after the fragrance transfer occurred, although VOC recovery was possible at higher temperatures and after longer persistence times. Given the transfer and persistence characteristics of VOCs from fragrance, there is potential for fragrance to be used as a form of trace in forensic reconstruction approaches.     

The impact of force,time, and rotation on the transfer of ammonium nitrate: A reductionist approach to understanding evidence dynamics

Empirical studies evaluating the conditions under which the transfer of forensic materials occurs can provide contextual information and offer insight into how that material may have been transferred in a given scenario. Here, a reductionist approach was taken to assess the impact of force, time, and rotation on the transfer of an explosive compound. An Instron ElectroPuls E3000 material testing instrument was used to bring porous and non-porous surfaces adulterated with an ammonium nitrate into direct contact with a human skin analogue, controlling for the force of contact, duration of contact, and rotation applied during contact. Quantifiable amounts of ammonium nitrate were recovered from all of the recipient surfaces demonstrating that ammonium nitrate is readily transferred from one surface to another, even when contact occurs for a short duration with a relatively low force. More particulates were transferred from non-porous surfaces onto the human skin analogue, but the amount of ammonium nitrate transferred did not depend upon the force of contact, duration of contact, or the amount of rotation applied. However, when contact occurred and involved rotation, a greater transfer of ammonium nitrate was observed, compared to those contacts occurring without rotation being applied. This approach complements more commonly-used holistic experiments that test multiple interacting variables in a realistic setting by isolating these variables, allowing them to be examined individually. This can be utilised to better understand the individual impact that specific variables have on the transfer of trace evidence in relevant crime reconstruction contexts.     

Visualising DNA transfer: Latent DNA detection using Diamond Dye

A central question is ‘how did DNA get there’? To help answer this, we visually monitored and recorded DNA transfer from one substrate to another. When an individual touches a substrate, traces of their DNA are transferred (primary/direct) which can then subsequently be transferred to a second substrate (secondary/indirect). Currently DNA transfer and how much remains can only be determined by collecting the biological material from the substrate, isolating the DNA and quantifying the amount recovered. However, Diamond™ Dye (DD) enables such DNA transfer events to be visualised by monitoring the movement of cellular material.We examined primary and secondary DNA transfer using aluminium as a primary substrate with cotton, polyester, aluminium and plastic as secondary substrates and four contact types between two substrates (passive, pressure, friction and friction with pressure). Participants pressed their index finger against the aluminium for 15 s and then DD was applied to the area of contact; cellular material was detected via a fluorescence microscope. Contact between that substrate and a second substrate was performed, using one of the four contact types. After this contact between substrates each was viewed microscopically and transfer of cellular material was recorded.Cellular material could be recorded as having transferred from one substrate to another. Substrate and contact type had an effect on the extent DNA transfers. DNA transferred at a high rate with aluminium as a primary substrate and cotton, polyester and aluminium as secondary substrates when pressure with friction was applied. This information expands our understanding of how DNA transfers and which factors affect it, thus assisting greatly with activity level reporting as to how DNA came to be where it was found.     

The transfer of touch DNA from hands to glass, fabric and wood

The transfer of DNA from hands to objects by holding or touching has been examined in the past. The main purpose of this study was to examine the variation in the amount of DNA transferred from hands to glass, fabric and wood. The study involved 300 volunteers (100 for glass, 100 for fabric and 100 for wood) 50% of which were male and 50% female. The volunteers held the material for 60 s. The DNA was recovered from the objects using a minitape lift, quantified using the Quantifiler kit assay, extracted using a ‘Qiagen^® QIAamp DNA mini kit’ and amplified using the AmpFlSTR^® SGM Plus™ Amplification Kit at 28 cycles. The results show that using ANOVA there was a significant difference (F = 8.2, p < 0.05) between the three object types in the amount of DNA recovered. In terms of DNA transfer and recovery, wood gave the best yield, followed by fabric and then glass. The likelihood of success of obtaining a profile indicative of the holder was approximately 9% for glass samples, 23% for fabric and 36% for wood. There was no significant difference between the amount of DNA transferred by male or female volunteers. In this study good shedder status, as defined by obtaining useful profiles of 6 or more alleles, is estimated at approximately 22% of the population. The phenomenon of secondary transfer was observed when mixed DNA profiles were obtained but the incidence was low at approximately 10% of the total number of samples. DNA profiles corresponding to more than one person were found on objects which had been touched by only one volunteer. Although secondary transfer is possible the profiles obtained from touched objects are more likely to be as a result of primary transfer rather than a secondary source.     

Transfer of DNA without contact from used clothing,pillowcases and towels by shaking agitation

DNA is frequently retrieved from commonly used objects or surfaces with no apparent biological stains. This DNA may have come from one or more individuals who directly deposited their DNA, or indirectly transferred the DNA of others, when physically contacting the sampled object or surface. Furthermore, contactless indirect DNA transfer of this ‘touch DNA’ from fabric substrates was recently demonstrated to be possible in a controlled laboratory environment. The circumstances and extent to which this form of contactless DNA transfer occurs are largely unknown. This study investigated indirect DNA transfer without contact by applying a gentle shaking agitation to used clothing, pillowcases and towels, with known usage and history, of 10 volunteers above the collection zone of the secondary surface. DNA transfer frequently occurred and was possible from all three investigated items. It occurred at levels that often produced informative profiles where transferred profiles closely resembled the profiles generated from the primary item. The outcomes of this study contribute to expanding the understanding of indirect DNA transfer without contact. However, this field would benefit from investigating a wider range of agitations and/or item types with various histories of use to determine the level of transfer and its detectability under different conditions.     

Evaluation of a Visualization Assay for Blood on Forensic Evidence

In forensics, bloodstains on dark fabrics might be invisible for the naked eye. Although several visualization, presumptive, and confirmatory blood tests have been developed, all have one or more disadvantages, especially on DNA analysis. We report here the use of a visualization assay that can visually detect blood drops up to 1/20 dilution. In this assay, the fabric is placed between two wet filter papers and covered by glass surfaces on both sides. Pressure is applied on the glass surfaces in which bloodstains transfer onto the filter papers through capillary forces. Detected stains can be tested with other more sensitive presumptive blood tests performed on the filter paper. Even more, DNA analysis can be performed on the transferred bloodstains. The presented visualization assay is easy to perform, extremely cheap, requires little hands on time, and does not affect bloodstain pattern analysis.     

Trace evidence dynamics of cocaine on banknotes: A comparison study of paper and polymer banknotes

It is well established that a large proportion of paper banknotes in circulation contain traces of cocaine. Being able to discriminate between the innocent transfer of illicit drug particles acquired through everyday interactions with surfaces such as banknotes, as opposed to transfer resulting from criminal activities can provide valuable intelligence that can inform an investigation. With many countries adopting polymer banknotes as legal tender, it is important to consider the transfer of cocaine from these surfaces as well as the retention of these particulates on polymer banknotes for evaluative interpretation in crime reconstruction. This comparison study assessed three contact variables (force, time, and rotation) on the transfer of cocaine particulates from paper and polymer banknotes onto a human skin proxy. The persistence of cocaine particulates was assessed through a realistic scenario which mimicked a cash transaction. Quantifiable amounts of cocaine were transferred onto the human skin proxy across all of the contacts assessed, with a greater transfer observed with contacts involving polymer banknotes and those contacts which involved rotation. Following extensive handling, cocaine persisted on both banknote types, with paper banknotes retaining larger amounts of cocaine than polymer banknotes. These findings show that cocaine can persist on both paper and polymer banknotes for extended periods of time following handling and is therefore available for transfer. This transfer then readily occurs, even when contact is brief and involves relatively small forces. A key distinction between the banknote types was that cocaine particulates are more likely to transfer from polymer banknotes due to the lower retention rate of particulates on this surface. Such insights can aid in evaluating the relevance of illicit drug particles identified on items or persons of interest in crime reconstruction approaches.     

Is it possible to predict the origin of epithelial cells? – A comparison of secondary transfer of skin epithelial cells versus vaginal mucous membrane cells by direct contact

In alleged sexual assault and rape cases, the focus has often been to collect samples from the victim's body, for detection of body fluids or skin cells from the offender. But in many cases intimate body samples from the perpetrator(s) can also be informative. However, in cases where the female victim claims vaginal penetration, the defendant may display an alternative explanation to the DNA findings, i.e. that the victim’s skin cells has been secondarily transferred to his penis. We hypothesized that female DNA will be detected in a significantly greater amount on swabs from penis after intercourse than after secondary transfer by skin contact.Fourteen male-female couples were recruited to test the above hypothesis, by collecting penile swabs from 3 specified anatomical locations: Glans, shaft, and the coronal sulcus, after two different situations: Vaginal intercourse and secondary transfer of epithelial cells by skin contact. The results show that penile swabs following intercourse produce significantly higher DNA concentrations than after secondary transfer by skin contact. Our results, indicates which of the anatomical regions is best suited for sampling. The DNA profiling results show a preponderance of female profiles over male profiles following intercourse compared to secondary skin contact.Based on these data, it is possible to make a statistical model to distinguish between samples taken after intercourse and samples taken after secondary transfer by skin contact based on the amount of female DNA and mixture proportion (Mx) between female and male DNA in samples collected from penis swabs.     

An innovative transfer DNA experimental design and qPCR assay to identify primary,secondary, and tertiary DNA transfer

“Touch DNA” is a form of trace DNA that is presumed to be deposited when an individual touches something and leaves behind DNA-containing skin cells, sweat, or other fluids. While touch DNA is often the result of direct contact (i.e., primary transfer), it can also be indirectly transferred between surfaces or individuals (e.g., secondary or tertiary transfer). Even experts cannot distinguish between different types of transfer and do not fully understand which variables affect direct versus indirect transfer or how often each type of transfer occurs. In this study, we utilize an innovative protocol that combines a paired male and female transfer DNA experimental design with an Amelogenin qPCR assay to generate data on primary, secondary, and tertiary DNA transfer. We report frequencies of indirect DNA transfer and also investigate the potential effects of participant age, self-identified ethnicity, and skin conditions on DNA transfer. Out of 22 experimental trials, we detected primary transfer (male + female) in 71% of trials, secondary DNA transfer in 50% of trials, and tertiary DNA transfer in 27% of trials. No significant associations were found between primary DNA transfer and age, self-identified ancestry, or skin conditions, however, all individuals with sloughing skin conditions demonstrated primary DNA transfer and we suggest this variable be explored in larger samples. These results contribute to a better understanding of the conditions under which secondary and tertiary DNA transfer occurs and can be used to propose realistic DNA transfer scenarios in court cases.     

An investigation on the secondary transfer of organic gunshot residues

Gunshot residues (GSR) are an important forensic trace in firearm-related events. Currently, routine GSR analyses focus on the detection and characterisation of the inorganic components (IGSR). The increasing prevalence of heavy metal-free ammunition challenges these current protocols and there is an increasing interest in how the organic components of GSR (OGSR) can provide complementary information. Similar to the situation with IGSR, OGSR compounds originally deposited on the shooter during the firing process may further be transferred onto another individual or surface. Hence, the aim of this study was to provide additional information regarding the risk of a secondary transfer of OGSR. Two scenarios were investigated, the first one related to the arrest process and the possibilities of a secondary transfer arising between a shooter onto a non-shooter (e.g. between a police officer and a person of interest (POI)). The second scenario concerned the transfer of OGSR onto the non-shooter after handling a firearm for few minutes without discharging it. One calibre was chosen, the .40 S&W calibre, used by several Australian State police forces. A secondary transfer was observed in all cases for the two scenarios investigated, for three compounds of interest: ethylcentralite (EC), diphenylamine (DPA), N-nitrosodiphenylamine (N-nDPA). The firearm handling scenario resulted in a larger secondary transfer to that of the arrest scenario. Overall, the amounts of OGSR detected on the non-shooter were generally lower than that detected on the shooter and controls after the arrest scenario. The results of this study provide complementary knowledge about OGSR, which can be further used to improve the current practice and the interpretation of OGSR evidence. In particular, it highlights that the secondary transfer proposition must be considered during the interpretation of forensic findings, especially when small amounts of OGSR target compounds are detected.     

Investigation into "normal" background DNA on adult necks: implications for DNA profiling of manual strangulation victims

Others have investigated the role that DNA profiling could play as a method for identifying the perpetrator of manual strangulation. These studies have demonstrated that it is possible to collect offender DNA from the skin surface of a victim following physical contact. It is not known whether nonself biological material is normally present on the skin surface due to adventitious transfer occurring during innocent everyday interactions. To test the hypothesis that detectable amounts of nonself DNA are normally present on the skin surface of healthy adult individuals due to the adventitious transfer of DNA occurring during normal day-to-day social interactions, we designed an experiment in three phases. Phase 1 was used to deduce which DNA collection, extraction, and amplification methods were suited to investigating this question. During phase 2, the neck surface of 24 healthy adult volunteers was swabbed. DNA was extracted using the QIAamp DNA mini kit and amplified using the SGM Plus PCR amplification kit, using 28 PCR cycles. The work carried out during phase 3 involved a simulated assault to investigate primary and secondary transfer of DNA during physical contact. It was found that 23% of neck areas swabbed during phase 2 of this investigation showed nondonor alleles in the resulting DNA profile, with 5% of areas showing six or more nondonor alleles. The results of phase 3 showed that primary, secondary, and zero transfer of victim and/or offender DNA could be observed after physical contact and that alleles from an unknown source could still be detected in this more controlled experiment. The data presented in this paper demonstrate that DNA profiles generated after swabbing the skin surface of healthy adults can include components of an unknown source, present due to adventitious transfer. These components, if present in large quantities, have the potential to interfere with DNA profile interpretation of swabs taken for the investigation of physical assault by DNA profiling.     

Impact Spatter Bloodstain Patterns on Textiles

There are few reports of studies of impact spatter on textiles even though bloodstained textiles are found at many violent scenes. Impact spatter was deposited at 90° impact angle onto three knit fabrics of different yarn sizes and on paper. The resulting stain areas and number of stains were measured using ImageJ and compared with stains on paper using one‐factor ANOVA. The number of stains observed and their areas on the knit fabrics decreased as the yarn size increased. It was also found that blood that deposited on the fabric wicked only in the direction of the fibers at that location within the fabric which led to distorted stain shapes. Fewer observed impact spatter stains were found on cotton jersey knits for fabrics made with larger yarns than on paper. As the yarn size became smaller, the number of stains became the same as on paper.     

An innovative transfer DNA experimental design and qPCR assay: Protocol and pilot study

Forensic “touch” DNA samples are low-quantity samples that are recovered from surfaces that have been touched by single or multiple individuals. These samples can include DNA from primary contributors who directly touched the surface, as well as secondary contributors whose DNA was transferred to the surface through an intermediary. It is difficult to determine the type of transfer, or how often and under what conditions DNA transfer occurs. In this paper, we present an innovative protocol that combines (1) a paired male and female transfer DNA experimental design in which the presence of male DNA indicates secondary transfer and (2) a cost-effective quantitative PCR (qPCR) assay of a sex-specific region in the Amelogenin gene to detect male and female DNA. We evaluate the ability of the Amelogenin qPCR assay to detect low concentrations of male and female DNA in mixed samples. We also test experimental DNA samples using our transfer DNA protocol to differentiate primary and secondary DNA transfer. Male DNA was detected in the majority of known mixed samples, even in samples with 4× more female DNA—this result demonstrates the ability to detect low concentrations of male DNA and the presence of secondary transfer DNA in our experimental design. Primary DNA transfer was detected in 100% of our experimental trials and secondary DNA transfer was detected in 37.5% of trials. Our innovative protocol mimics realistic case scenarios to establish rates of primary and secondary DNA transfer in an inexpensive and simplified manner.     

Improving the effectiveness of fluorescence for the detection of semen stains on fabrics

The paper describes the use of the Polilight, a light source based on a xenon arc lamp, to exploit the fluorescence properties of semen as an aid to searching fabrics for stains in sexual assault cases. The broad excitation spectrum of semen allows the fluorescence to be generated at a range of wavelengths. This permits the excitation and emission conditions to be selected that minimize interference from background fluorescence of the fabric and thereby optimizes the contrast between the fabric and the stain. A common method for the observation of fluorescence is the use of colored plastic goggles or filters. The paper shows that the detection of fluorescence from semen stains is significantly enhanced using appropriate interference filters.     

Tearing of knicker fabrics

In Dunedin (South Island, New Zealand), a sexual assault is reported to police approximately once every two to three weeks, with some reports fictitious. Identifying a fictitious claim is difficult, and damage to apparel, especially knickers, may be the only form of evidence. In this paper, the tear behaviour of three knit fabrics, typical of those used to manufacture knickers is reported: the effect of laundering prior to tearing was considered. Tearing behaviour was determined using an Instron universal testing machine (Model 4464) operating in tensile mode to eliminate variability which is inevitable with human participants. Cotton and cotton-rich fabrics were more difficult to tear than modal-rich fabrics: the addition of elastane increased the time for the tear initiation as elastane fibres allowed the fabric to extend more before breaking. Specimens behaved differently depending on which direction they were torn (course-direction specimens down the length of the specimen, wale-direction specimens 50% down the length, 50% across the specimen). Laundered fabrics required less force to tear than new fabrics, therefore, when examining torn apparel, the fibre content and age of the garment need to be considered. Torn fibre ends appeared similar to those damaged by other means (e.g. knife, screwdriver) and no features visible under FESEM could be attributed solely to tearing damage in the fabrics studied.     

Assessing the potential fire risk of laundered fabrics after contamination with emollients using ATR-FTIR spectroscopy and chemometrics

Since 2010, more than 50 UK fire deaths, have been reported as linked with emollients. This prompted the Medicines and Healthcare products Regulatory Agency (MHRA) to issue advice on their safer use in 2018. The advice was in response to concerns raised by the National Fire Chiefs Council, coroners’ reports, and flammability tests. The test results show a significant reduction in ignition time of fabrics contaminated with paraffin-based and paraffin-free skin care product residues. The MHRA report also included advice on washing clothing and bedding at high temperatures but warned this may not remove all emollient residues. This paper reports on new research on the removal of skin care products from clothing investigated by laundering contaminated 100% cotton fabric at 30, 40 and 60 °C using both biological and non-biological based detergents. As part of the experiment, non-contaminated (blank) napkin samples were included in the wash experiments to assess the possible transfer from fabrics contaminated with emollients to uncontaminated clean fabrics during washing. Washed and dried fabrics were analysed using Attenuated Total Reflectance, Fourier Transform Infrared (ATR-FTIR) spectroscopy and further interpreted via principal component analysis (PCA) and network analysis. Results suggest that the majority of the 6% white soft paraffin-based lotion and paraffin-free cream were removed at all temperatures. Residues of 21% paraffin-based cream (6% light paraffin/15% white soft paraffin) remain, and more residues persist of the 100% paraffin-based ointment (5% light paraffin/95% white soft paraffin) after washing at 30, 40 and 60 °C. The wash experiments show unequivocal transfer of the 100% ointment from the contaminated napkins to clean control napkins placed within washes at 30 °C. Furthermore, residues of the ointment were observed within the machine drum, and washing machine door seal, though this did not cause secondary transfer to subsequent wash experiments. There were no differences observed when using biological versus non-biological detergents, nor when employing a pre-wash treatment in the removal of residues of the 21% cream and 100% ointment. These results suggest that a single application of an emollient when soaked and dried into a fabric is not removed by a single wash at 30, 40 or 60 °C. Instead, the residue remains a persistent potential fire risk and, its high paraffin content presents an additional fire risk via contamination of other fabrics.     

The killer outfit and timing: Impact of the fabric and time in body fluid identification and DNA profiling

The present work aimed to study the detection, through lateral flow immunochromatographic (LFI) tests, of saliva samples over time in three different types of fabrics, as well as, the possibility of DNA isolation and characterization from the sample tubes and the cassettes. Fifty microliters of saliva (three samples/time) were deposited in denim, cotton, and polyester. Saliva was identified by SERATEC® Amylase Test and the Crime Scene version SALIVA CS, being able to detect it up to six months of deposition, although with different band intensities. Polyester showed stronger bands than cotton, probably due to its synthetic nature, and denim, as an inked fabric, showed less band intensities. Statistical analyses confirmed significant differences among fabrics, but not over time in the same type of fabric. Total DNA from the sample tubes was successfully recovered, in contrast, from the cassettes, only polyester retrieved amplifiable DNA. These findings indicated that it is possible to recover and identify saliva up to six months after deposition, also obtaining DNA. Future research will be able to expand these results, analyzing the stability of other body fluids, and the sensitivity of lateral flow immunochromatographic tests to detect them.     

Could Secondary DNA Transfer Falsely Place Someone at the Scene of a Crime?,

The occurrence of secondary DNA transfer has been previously established. However, the transfer of DNA through an intermediary has not been revisited with more sensitive current technologies implemented to increase the likelihood of obtaining results from low‐template/low‐quality samples. This study evaluated whether this increased sensitivity could lead to the detection of interpretable secondary DNA transfer profiles. After two minutes of hand to hand contact, participants immediately handled assigned knives. Swabbings of the knives with detectable amounts of DNA were amplified with the Identifiler^® Plus Amplification Kit and injected on a 3130xl. DNA typing results indicated that secondary DNA transfer was detected in 85% of the samples. In five samples, the secondary contributor was either the only contributor or the major contributor identified despite never coming into direct contact with the knife. This study demonstrates the risk of assuming that DNA recovered from an object resulted from direct contact.     

A model for estimating the number of glass fragments transferred when breaking a pane: experiments with firearms and hammer.

Numerous publications have shown the importance of transfer in the interpretation of glass evidence. As this phenomenon is also highly variable, it was decided to test the hypothesis that there exists a means to predict the number of fragments recovered at time t = 0. Panes of float glass-of different types and thickness-were broken using either a firearm or a hammer. It was decided to choose a firearm as the main breaking device, as it allowed not only to have more reproducible conditions but also to acquire knowledge in a field where little has been published. Despite the inherent variation in the breaking process, the results show that using a statistical model it is possible to predict the number of fragments transferred onto a garment from the number of fragments transferred to the ground. This research also indicates the size and number of particles transferred onto a person, when breaking window panes of different types (float, laminated or toughened) with different breaking procedures.     

A quantitative assessment of the extent and distribution of textile fibre transfer to persons involved in physical assault

Knowledge of the number of fibres transferred during a particular activity is essential for the interpretation of findings in similar criminal cases. In this regard, violent contacts and physical assaults still present a challenge, due to a lack of robust published data. Hereby, we present the outcome of an empirical study where different assault activities were simulated by a Jiu Jitsu team and participants were asked to play either the role of an aggressive ‘assailant’ or a defensive ‘victim’, wearing cotton garments (i.e., Gi’s). Four different scenarios were simulated in replicates (n = 5), each of them involving different intensity levels (low and high) and duration times (30 and 60 s). Results showed that approximately 1,000 to 44,000 fibres were cross-transferred between the participants’ garments, with noticeable differences between the different scenarios. These values were significantly larger than those published in previous studies and, therefore, suggested the possibility of a current underestimation of the number of fibres transferred in physical assaults. Furthermore, statistical analysis by ANOVA indicated that the all the variables tested (i.e., intensity level, duration time, and participants role) had a significant effect on the number of transferred fibres (p < 0.001) and, consequently, that some knowledge of the case circumstances may be important to make more educated estimations. This is the first time that such a methodology has been applied for the quantitative assessment of fibre transfer between participants in assault activities. Data are expected to help practitioners with the interpretation of findings in real casework and lead to a more robust evidential assessment.