The kinetics of colour change in textiles and fibres treated with detergent solutions: Part II — Spectrophotometric measurements

The aim of this study was to assess colour variations that occur in several types of textiles and their constituent fibres, resulting from the long-term influence of various laundry detergents. A 14-day experiment was conducted using blue, red and grey/black cotton, wool, acrylic and polyester textiles. The spectrophotometric measurement of colour changes in fabric samples and test solutions, as well as the microspectrophotometric analysis of colour changes in single fibres were described. An evaluation of the observed colour changes from a forensic fibre analysis expert's point of view, as well as that of an average user/consumer of the textiles and laundry detergents is also provided.     

Evaluation of two DNA/RNA co-extraction methods for body fluid identification in forensics

Body fluid identification has become a field of interest in forensic casework as it can add value to particular investigative scenarios. Identifying the source of the biological material is not always an upfront task using conventional methods; therefore, profiling of specific mRNA markers can provide the answer. The implementation of RNA based analyses in forensic casework must focus on the quality and sensitivity of methods, starting with nucleic acid extraction, and without loss of DNA for STR profiling. In this work, two methods for DNA and RNA co-extraction were tested and compared: a commercial kit that uses a spin, mini column methodology, and a quick, simple nucleic acid isopropanol precipitation based protocol. Both methods simultaneously extract DNA and RNA, crucial for forensic casework and were tested in semen samples. Nucleic acid quantifications as well as purity assessment ratios (OD₂₆₀/OD₂₃₀ and OD₂₆₀/OD₂₈₀) were obtained by both methods to infer on the use of extracts in downstream applications such as PCR. The performance of the two tested protocols was further evaluated by analyzing two semen mRNA specific markers, PRM1 and SEMG1. When compared to the commercially developed kit, results suggest that the literature adapted protocol allowed more carryover of contaminants absorbing at 230 nm and 280 nm as the purity ratios were below the accepted standard ranges. Negative results for mRNA profiling supported the QC results obtained by spectrophotometry. On the hand, PRM1 and SEGM1 were positive in RNA samples extracted with the commercial kit.     

An investigation into the persistence of textile fibres on buried carcasses

A significant amount of research has been carried out on fibres to aid the forensic scientist in determining the significance of these when found on a victim or suspect. This work has focused on open-air environments, and as such no research has been undertaken to examine the persistence of fibres on bodies in the burial environment.Wool and cotton fibres, known to fluoresce under ultraviolet (UV) light, were transferred onto the skin of four porcine (Sus scrofa) carcasses (two carcasses per fibre type). The number of fibres transferred was recorded from images taken under UV light. The remains were subsequently placed in four burial sites and left interred for 14 days. After this period the carcasses were excavated and lightly brushed down to remove the soil layer that had adhered to the skin. Once again photography under UV light was used to record the number of fibres which persisted on the skin.Results showed that after 14 days, wool and cotton fibres remain on the surface of the buried carcasses. In no circumstance was there a total loss of fibres suggesting that in such scenarios, the likelihood of finding fibres is high but the initial number of fibres transferred would be strongly diminished. This has important implications for both the excavation protocol for buried remains and the subsequent analysis for physical evidence.     

Forensic DNA laboratory automation – Principles and guidelines

There is a lack of clear guidelines for project managers, laboratory managers and forensic scientists on strategies for the automation of forensic DNA laboratory processes and operational implementation of new technologies. This is reflected in the failure rate of projects in the forensic DNA testing environment. We present a set of guidelines and concepts important for forensic laboratory automation. Some case studies from past projects are presented. These consist of partial (or modular) automation (n = 2) and full automated robotically integrated systems (n = 2).Technology Management principles and concepts are crucial to prevent failure of projects, e.g. early adoption of untried technologies, and organizational factors. The future of laboratory automation is modular until such time as new discontinuous technologies will replace the need of the traditional manual laboratory configuration in totality.     

The effect of re-annealing on the distribution of refractive index in a windscreen and a windowpane classification of glass samples

Tiny glass fragments of two typical objects being of interest in forensic investigations: car windscreen and windowpane were examined from the point of view of their importance as crime evidence. Both examined objects were made from float glass and were toughened. The present paper concerns examination of refractive index distribution across the objects under investigation before and after the glass fragments were annealed according to previously chosen procedure. The annealing procedure was carried out in order to increase discrimination power of refractive index (RI). The following conclusions can be drawn from the results obtained. For both examined objects the mean RI was significantly higher after annealing and, at the same time, standard deviations in RI were smaller. The distributions of RI for both examined objects appeared not to be normal; the deviations from normality were observed at both sides of RI distributions. It was found that the difference in the values DeltaRI (difference between mean values of RI after annealing and before annealing) for both examined object was not significant and thus it would be not a good parameter to differentiate between two heat strengthen objects. The attempt to classify 181 glass samples on the basis of theirs RI and DeltaRI was performed. Increased discrimination of glass samples was observed.     

油浸法鉴别汽车后视镜玻璃的研究

目的研究油浸法对汽车后视镜玻璃的鉴别能力。方法应用油浸法对20种汽车后视镜玻璃进行折射率值测定并用t检验法对数据进行处理。结果在置信度为95％时，20个后视镜玻璃样品的区分率为97．9％。结论油浸法对汽车后视镜玻璃有很好的区分能力，而且使用样品量少，可以达到微克级，是一种鉴别后视镜玻璃的有效方法。     

A More Efficient Method for Synthetic Textile Fiber Analysis Using Polarized Light Microscopy

The efficiency of conventional polarized light microscopy (PLM) methods for analyzing synthetic fiber evidence analyses is improved. Historically, using PLM for fiber identification relied on measuring refractive index. This prior PLM technology is reliable, but it is not efficient. Most fibers are optically anisotropic, having two principal refractive index values, N_(High) and N_(Low). When the fiber is mounted in intermediate refractive index medium, efficiency is improved by observing the change in contrast while the polarized light’s vector is rotated relative to the fiber’s axis. Minimum contrast occurs when the refractive indices of the mounting medium and fiber are equal. This angle of equality is determined by orienting the fiber’s highest refractive index parallel to the polarized light’s electric field vector, rotating the fiber or polarizing element, observing minimum contrast and measuring the angle of equality. This method is rapid, reduces remounting fibers in different mounting media and provides a quantitative measure for fiber comparisons.     

Effectiveness of Single‐Nucleotide Polymorphisms to Investigate Cattle Rustling

Short tandem repeats (STR)s have been the eligible markers for forensic animal genetics, despite single‐nucleotide polymorphisms (SNP)s became acceptable. The technology, the type, and amount of markers could limit the investigation in degraded forensic samples. The performance of a 32‐SNP panel genotyped through OpenArrays^TM (real‐time PCR based) was evaluated to resolve cattle‐specific forensic cases. DNA from different biological sources was used, including samples from an alleged instance of cattle rustling. SNPs and STRs performance and repeatability were compared. SNP call rate was variable among sample type (average = 80.18%), while forensic samples showed the lowest value (70.94%). The repeatability obtained (98.7%) supports the used technology. SNPs had better call rates than STRs in 12 of 20 casework samples, while forensic index values were similar for both panels. In conclusion, the 32‐SNPs used are as informative as the standard bovine STR battery and hence are suitable to resolve cattle rustling investigations.     

Axonal injury – a diagnostic tool in forensic neuropathology?: A review

We used β-amyloid precursor protein (β-APP) to investigate our own forensic neuropathological case material (n=252) in light of the current literature on the phenomenon “axonal injury” (AI) to determine the incidence, specificity and biomechanical significance of AI and its significance for determining vitality and survival time. The case material consisted of cases of fatal nonmissile closed-head injury (n=119), gunshot injury (n=30), fatal cerebral ischemia/hypoxia (n=51), brain death caused by mechanical trauma (n=14) or nonmechanical injury (n=18), and acute hemorrhagic shock (n=20). AI was observed in 65% to 100% of cases of closed-head injury, fatal cerebral ischemia/hypoxia, and brain death with a survival time of more than 3 h; AI could not be detected in the cases of acute hemorrhagic shock. A statistically significant difference between traumatically and nontraumatically induced (nondisruptive) AI was not found. There was no statistical evidence of a correlation between AI and the different types of external force, since AI could be demonstrated after both acceleration/deceleration injuries and traumatic impact. Therefore, biomechanical inferences for reconstruction purposes are not possible. On the other hand, β-APP was found to be a definite marker of vitality. In our material, cases with a posttraumatic interval of under 180 min did not express β-APP. Moreover, the literature shows that the posttraumatic interval can be determined by other methods for demonstration of AI such as by ubiquitin immunostaining (360 min), silver staining (15–18 h), hematoxylin and eosin staining (about 24 h), or by demonstration of a microglial reaction (about 4 to 10 days) or of a few remaining isolated bulbs, without accompanying fibers, which can be detected after a survival time of up to 17 months.     

If error rate is such a simple concept,why don’t I have one for my forensic tool yet?

The Daubert decision motivates attempts to establish error rates for digital forensic tools. Many scientific procedures have been devised that can answer simple questions. For example, does a soil sample contain component X? A procedure can be followed that gives an answer with known rates of error. Usually the error rate of a process that tries to detect something is associated with a random component of some measurement. Typically there are two types of error, type I, also called a false positive (detecting it when it is not really there), and type II, also called a false negative (missing it when it really is there). At first thought, an error rate for a forensic acquisition tool or a write blocking tool is a simple concept. An obvious possibility for the error rate of an acquisition is k/n, where n is the total number of bits acquired and k is the number of incorrectly acquired bits. However, the kinds of errors in the soil test and in digital acquisition are fundamentally different. The errors in the soil test can be modeled with a random distribution that can be treated statistically, but the errors that occur in a digital acquisition are systematic and triggered by specific conditions. The purpose of this paper is not to define any error rates for forensic tools, but identification of some of the basic issues to stimulate discussion and further work on the topic.     

A data collection of vehicle topcoat colours. 2. The measurement of colour samples used in the vehicle refinishing industry

Four collections of painted cards, of the type used as reference samples in the vehicle refinishing industry, have been examined for suitability for incorporation in a data base of vehicle topcoat colours. One of these was considered more suitable for forensic science applications than the others and tristimulus values (X,Y,Z) were obtained for over 6000 samples. The data were then mathematically transformed to a uniform colour space and the sample distribution was plotted according to three colour axes. The evidential value of the plots is briefly discussed.     

Assessing Carbon and Hydrogen Isotopic Fractionation of Diesel Fuel n-alkanes During Progressive Evaporation

Compound-specific isotope analysis offers potential for fingerprinting of diesel fuels, however, possible confounding effects of isotopic fractionation due to evaporation need to be assessed. This study measured the fractionation of the stable carbon and hydrogen isotopes in n-alkane compounds in neat diesel fuel during evaporation. Isotope ratios were measured using a continuous flow gas chromatograph/isotope ratio mass spectrometer. Diesel samples were progressively evaporated at 24 ± 2°C for 21 days. Increasing depletion of deuterium in nC₁₂–nC₁₇ alkanes in the remaining liquid with increasing carbon chain length was observed. Negligible carbon isotope fractionation was observed. Preferential vaporization was measured for the shorter chain n-alkanes and the trend decreased with increasing chain length. The decrease in δ²H values indicates the preferential vaporization of the isotopically heavier species consistent with available quantitative data for hydrocarbons. These results are most important in the application of stable isotope technology to forensic analysis of diesel.     

Multiplex-direct PCR assay for foodborne pathogen identification: An application in forensic investigation

Foodborne pathogens present serious concerns to human health and can even lead to fatalities. Microbial forensic science thus plays an important role in consumer protection, food security, and even in litigation. The gold standard for pathogen identification – bacterial culture – is costly and time-consuming. A cheaper and quicker alternative will benefit both forensic science and medical diagnosis. In this study, we developed and validated a molecular-based method termed ‘multiplex-direct PCR assay’ to simultaneously detect three common foodborne pathogens – Escherichia coli O157:H7, Campylobacter jejuni, and Listeria monocytogenes. Three previously reported species-specific primer pairs were modified and used to directly amplify samples without DNA extraction. The assay was also validated for its specificity, sensitivity, and applied to test several samples obtained from a local market and clinical samples. The results showed the expected PCR fragments of approximately 490, 343, and 209 bp for E. coli O157:H7, C. jejuni, and L. monocytogenes, respectively. The assay was specific to the targeted pathogens and was sufficiently sensitive and robust to effectively analyze market samples. The whole process took less than 1 h to complete indicating that the assay is suitable for reliable, rapid, and inexpensive identification of these three foodborne pathogens, which could be useful in microbial forensic investigation.     

Population data for six X-chromosome STR loci in a Rio de Janeiro (Brazil) sample: Usefulness in forensic casework

This study presents data for the X-chromosome STR loci DXS7133, DXS7424, DXS8378, DXS6807, DXS7423 and DXS8377. In order to establish a database, unrelated individuals (males and females) from Rio de Janeiro were typed for the above loci. No significant differences were observed between allele frequencies in male and female samples (non-differentiation exact P values ≥ 0.156). Hardy-Weinberg equilibrium was tested in the female sample and no significant deviations were found. All six markers have shown to be highly polymorphic in our sample with gene diversities varying between 0.6797 for DXS7133, and 0.9260 for DXS8377. Pairwise linkage disequilibrium analysis did not allow discharging a possible association between DXS7133 and DXS7424 alleles in Rio de Janeiro population. Parameters of forensic interest, like PD_M, PD_F, Het_obs, Het_exp, were calculated for each locus. The high discrimination power estimated in both males and females, as well as mean exclusion chance in father/daughter duos and in father/mother/daughter trios, demonstrates the usefulness of these six markers in forensic investigation.     

Sequence variation at three X chromosomal short tandem repeats in Caucasian and African populations

Sequence variation for the X chromosome short tandem repeats (X-STRs) DXS9898, DXS6789 and GATA172D05 was studied in two major population groups, namely Caucasians and Africans. DXS6789 revealed two different subtype sequence polymorphisms: for shorter alleles, with less than 17 repeats, results showed a simple composition with the following structure: (TATG)_m-(TATC)_n. For longer alleles, a constant TATC insertion was observed at the beginning of the variable repeat unit. Additionally, alleles identical in size revealed structural variations regarding the TATG/TATC proportion. Africans showed a higher intra-allelic variation at this locus than the Caucasian population group. For all three loci, DXS9898, DXS6789 and GATA172D05, no unique structure was found among Africans and Caucasians.     

Variations in vitreous humor chemical values as a result of instrumentation

Urea nitrogen, glucose, sodium, potassium, and chloride were measured in common vitreous humor samples using a variety of instruments. There was found to be variation in values obtained by the different procedures for each of these constituents. The variation in electrolyte values between the different procedures can pose real problems in attempting to determine the presence of an antemortem dehydration or low salt condition. Possible reasons for these variations are discussed, and the normal range of values of both sodium and chloride for the different instrumentalities is provided. However, variations in values for both urea nitrogen and glucose would not pose any problems of interpretation for forensic science evaluations.     

Analysis of genetic polymorphisms associated with the presence of freckles for phenotypic prediction

The prediction of externally visible characteristics (EVCs) is a commonly used practice by the forensic sciences as an important resource in the investigation of criminal cases in which the identity of perpetrators or victims is unknown or even to recognize decomposed cadavers. With this purpose, genetic markers associated with pigmentation traits have been widely studied by forensic scientists and, nowadays, it is possible to predict phenotypic characteristics such as hair, eyes and skin colour, as well as the presence of skin freckles by analysing single nucleotide polymorphisms (SNPs). In this study, we analysed the association of six SNPs located in pigmentation genes to the presence of freckles in individuals from the Brazilian population for forensic DNA phenotyping. The study was based within the context of a larger project on a population sample of 534 adult Brazilians of both sexes and different skin colours. DNA was extracted from peripheral blood and genotyped using the TaqMan® OpenArray® Real-Time PCR System (ThermoFischer Scientific) technique. Statistical analyses were carried out with the R software (version 4.0.2). As for the results obtained, three SNPs were shown to be statistically associated to the freckling, rs12203592, rs1800404 and rs222847, with CT, AG and AA genotypes being the main contributors, respectively. Variables such as sex of the individuals and skin colour were found to also contribute to the manifestation of this pigmentation trait. Further statistical analyses will be carried out to evaluate the possibility of using the SNPs in this study for phenotyping prediction of the Brazilian population, improving existing DNA phenotyping models in forensic sciences.     

Evaluation of neodymium isotope analysis of human dental enamel as a provenance indicator using 1013 Ω amplifiers (TIMS)

Human provenance studies employing isotopic analysis have become an essential tool in forensic and archaeological sciences, with multi-isotope approaches providing more specific location estimates compared to single isotope studies. This study reports on the human provenancing capability of neodymium isotopes (¹⁴³Nd/¹⁴⁴Nd), a relatively conservative tracer in the environment. Neodymium isotope ratios have only recently been determined on human remains due to low concentrations in human dental enamel (ppb range), requiring thermal ionisation mass spectrometry (TIMS) using 10¹³ Ω resistors. Dental elements (third molars) from 20 individuals born and raised in the Netherlands were analysed for Nd concentration (n = 12) and Nd isotope ratios (n = 15). The geological control on Nd isotope composition was examined using coupled Nd-Sr isotope analysis of the same third molar. Teeth from different geological environments were also analysed (Caribbean, Columbian, and Icelandic, n = 5). Neodymium elemental concentrations in dental elements ranged between 0.1 and 7.9 ppb (median 0.5 ppb). The Dutch ¹⁴³Nd/¹⁴⁴Nd ratios of the provinces of Limburg and Friesland were between 0.5118 and 0.5121, with Dutch ⁸⁷Sr/⁸⁶Sr ratios in agreement with the previously established local range (0.708–0.710). The current findings were compared to previously published results on Nd concentration and composition from Dutch individuals. The concentration of Nd and ¹⁴³Nd/¹⁴⁴Nd ratios were weakly correlated (R² = 0.47, n = 17) in Dutch human dental enamel. The majority (n = 25, 83.3%) of individuals had Nd and Sr isotope values isotopically indistinguishable from the geological environment in which their third molars formed and mineralised. However, the Nd isotope ratios of the Icelandic individual and several Dutch individuals (n = 4) suggested that Nd in enamel is not solely influenced by geological environment. In order for neodymium isotopes to be quantitatively applied in forensic and archaeological settings further analyses of individuals from various geographical regions with well-defined dietary Nd isotope data are required.     

Uncertainty in Widmark calculations: ABV variation in packaged versions of the most popular beers in the UK

Forensic practitioners regularly use the Widmark equation to determine theoretical blood alcohol concentrations for use in cases involving alcohol. It is important with these calculations to determine the uncertainty associated with any result. Previous work has investigated the uncertainty in percent alcohol by volume (%ABV) from beers produced by small independent breweries in the UK but did not study the top selling beers in the UK. The top selling lagers and ales/bitters in the UK were identified by sales volume and the %ABV determined. These data was then used to determine the percent coefficient of variation (%CV) that should be used by forensic practitioners when constructing alcohol technical defence reports for use in forensic cases. These samples, from what may be described as ‘big’ brewers, were determined to have a smaller root mean square error (RMSE) (±0.1%v/v, n?=?35), and %CV than those previously reported for beers produced by small, independent breweries in the UK. The results from this study shows that different RMSE's should be used for %ABV when determining the uncertainty of results from Widmark calculations depending if the drinks consumed have been from either ‘big’ brewers or small, independent breweries.     

Cytochrome b based genetic differentiation of Indian wild pig (Sus scrofa cristatus) and domestic pig (Sus scrofa domestica) and its use in wildlife forensics

The Indian wild pig (Sus scrofa cristatus) is a protected species and listed in the Indian Wildlife (Protection) Act, 1972. The wild pig is often hunted illegally and sold in market as meat warranting punishment under law. To avoid confusion in identification of these two subspecies during wildlife forensic examinations, we describe genetic differentiation of Indian wild and domestic pigs using a molecular technique. Analysis of sequence generated from the partial fragment (421 bp) of mitochondrial DNA (mtDNA) cytochrome b (Cyt b) gene exhibited unambiguous (> 3%) genetic variation between Indian wild and domestic pigs. We observed nine forensically informative nucleotide sequence (FINS) variations between Indian wild and domestic pigs. The overall genetic variation described in this study is helpful in forensic identification of the biological samples of wild and domestic pigs. It also helped in differentiating the Indian wild pig from other wild pig races. This study indicates that domestic pigs in India are not descendent of the Indian wild pig, however; they are closer to the other wild pig races found in Asia and Europe.