Recovery of human DNA profiles from poached deer remains part 2: Improved recovery protocol without the need for LCN analysis

Although poaching is a common wildlife crime, the high and prohibitive cost of specialised animal testing means that many cases are left un-investigated. We previously described a novel approach to wildlife crime investigation that looked at the identification of human DNA on poached animal remains (Tobe, Govan and Welch, 2011). Human DNA was successfully isolated and amplified from simulated poaching incidents, however a low template protocol was required which made this method unsuitable for use in many laboratories. We now report on an optimised recovery and amplification protocol which removes the need for low template analysis.Samples from 10 deer (40 samples total — one from each leg) analysed in the original study were re-analysed in the current study with an additional 11 deer samples. Four samples analysed using Chelex did not show any results and a new method was devised whereby the available DNA was concentrated. By combining the DNA extracts from all tapings of the same deer remains followed by concentration, the recovered quantity of human DNA was found to be 29.5 pg ± 43.2 pg, 31 × greater than the previous study. The use of the Investigator Decaplex SE (QIAGEN) STR kit provided better results in the form of more complete profiles than did the AmpF?STR® SGM Plus® kit at 30 cycles (Applied Biosystems). Re-analysis of the samples from the initial study using the new, optimised protocol resulted in an average increase of 18% of recovered alleles. Over 17 samples, 71% of the samples analysed using the optimised protocol showed sufficient amplification for comparison to a reference profile and gave match probabilities ranging from 7.7690 × 10^? 05 to 2.2706 × 10^? 14.The removal of low template analysis means this optimised method provides evidence of high probative value and is suitable for immediate use in forensic laboratories. All methods and techniques used are standard and are compatible with current SOPs. As no high cost non-human DNA analysis is required the overall process is no more expensive than the investigation of other volume crime samples. The technique is suitable for immediate use in poaching incidents.     

Duplex-specific nuclease (DSN): A method for the rehabilitation of low-copy number DNA profiles

A continual challenge in the field of forensic DNA analysis is the amplification and interpretation of degraded and low-copy number (LCN) DNA obtained from amounts of limited biological evidence. It has been well established that DNA profiles obtained from the amplification of low quality, degraded, and/or LCN DNA samples are often of limited value due to the frequent occurrence of preferential amplification during polymerase chain reaction (PCR). The by-products of preferential PCR amplification are often observed as inter- and intra-locus peak imbalance, allelic dropout, and/or locus dropout. These are all artifacts that are identified during the interpretation phase of analysis rather than by improving the quality of the DNA present. While it is theoretically possible to obtain a complete DNA profile from a single cell, in reality, profiles obtained from suboptimal amounts of DNA are difficult to interpret and frequently inconsistent when replicated. Inspired by advances in next-generation sequencing techniques, we propose a methodology for simultaneously normalizing the abundance of PCR products across all short tandem repeat (STR) loci using the DNA exonuclease, duplex-specific nuclease (DSN). DSN is an enzyme isolated from the hepatopancreas of Red King (Kamchatka) crab that possesses a strong affinity for digesting double stranded DNA (dsDNA) and has limited activity toward single stranded DNA (ssDNA). Degraded DNA known to display peak imbalance and allele dropout was amplified using AmpFlSTR^® Identifiler^® Plus for 28 cycles. Following amplification, samples were denatured at 99.9 °C for 5 min and incubated with one unit of DSN at 62 °C in a 28 μl volume for 1 min. Nuclease activity was terminated through the addition of equal volume of 10 mM EDTA and 95 °C incubation for 2 min. Following DSN treatment, 21 of 30 alleles within the known profile exhibited some improvement in peak height balance. The findings obtained support the potential use of DSN treatment as a method for normalizing STR profiles and improving the quality of data from degraded and low quantity DNA samples.     

Research potential and limitations of trace analyses of cremated remains

Human cremation is a common funeral practice all over the world and will presumably become an even more popular choice for interment in the future. Mainly for purposes of identification, there is presently a growing need to perform trace analyses such as DNA or stable isotope analyses on human remains after cremation in order to clarify pending questions in civil or criminal court cases. The aim of this study was to experimentally test the potential and limitations of DNA and stable isotope analyses when conducted on cremated remains.For this purpose, tibiae from modern cattle were experimentally cremated by incinerating the bones in increments of 100 °C until a maximum of 1000 °C was reached. In addition, cremated human remains were collected from a modern crematory. The samples were investigated to determine level of DNA preservation and stable isotope values (C and N in collagen, C and O in the structural carbonate, and Sr in apatite). Furthermore, we assessed the integrity of microstructural organization, appearance under UV-light, collagen content, as well as the mineral and crystalline organization. This was conducted in order to provide a general background with which to explain observed changes in the trace analyses data sets. The goal is to develop an efficacious screening method for determining at which degree of burning bone still retains its original biological signals. We found that stable isotope analysis of the tested light elements in bone is only possible up to a heat exposure of 300 °C while the isotopic signal from strontium remains unaltered even in bones exposed to very high temperatures. DNA-analyses seem theoretically possible up to a heat exposure of 600 °C but can not be advised in every case because of the increased risk of contamination. While the macroscopic colour and UV-fluorescence of cremated bone give hints to temperature exposure of the bone's outer surface, its histological appearance can be used as a reliable indicator for the assessment of the overall degree of burning.     

Transfer of fibres on the hands of living subjects and their persistence during hand washing

Textile fibres were transferred to the hands of ten living subjects and their persistence was determined after hand washing. Average number of fibres transferred was 300 ± 133 (female 288 ± 92, male 311 ± 163) per 100 cm² hand area in the 100 experiments. However the number of fibres transferred was not gender dependent but individual dependent. The hand texture of subjects was compared with the number of fibres transferred but the relationship was not observed. The number of fibres transferred varied significantly for the 10 repeated experiments performed under the same conditions for the same subject.The subjects were then asked to wash their hands with water. One test group washed their hands with standing water, and the other with running tap water. Afterwards, the number of fibres remaining on the test subjects' hands were investigated. Migration of the fibres on the surface of the observed hands did occur but total loss of transferred fibre after hand washing did not occur. The average number of fibres remaining per 100 cm² hand area was 14 ± 10 (range = 3–72) for hand washing with standing water, and 10 ± 12 (range = 0–79) for washing with running tap water. The results of this study show the possibility of finding fibres on the hands of a person involved in a criminal case even after hand washing before fibre collection.     

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.     

Suspect burial excavation procedure: A cautionary tale

Geographic location, time of reporting and need for rapid evaluation contributed to a lack of intelligence concerning a suspect burial site in scrub woodland (～15 km from the last known location of a missing person) in Northern Ireland. Police received reports of a subsiding ‘grave’, which was evaluated positively using GPR and victim recovery dogs (VRD). After 24 h work, archaeological excavation showed a vertical-sided, stepped excavation on undisturbed clay with no inhumation. Subsequent research showed the feature to be an engineering trial pit. The GPR response was a water table and rocks, VRD were possibly reacting to disturbed ground. The work serves as a demonstration of good archaeological practice in suspect burial excavation, following a lack of landscape evaluation and poor overall intelligence.     

Characterisation of forward stutter in the AmpFlSTR® SGM Plus® PCR

PCR amplification of tetrameric short tandem repeats (STRs) can lead to Taq enzyme slippage and artefact products typically one repeat unit less in size than the parent STR. These back stutter or n ? 4 amplification products are low-level relative to the amplification of the parent STR but are widely seen in the forensic community where tetrameric STRs are employed in the generation of DNA profiles. To aid the interpretation of DNA mixtures where minor contributor(s) might be present in comparable amounts to the back stutter products, the typical amounts of back stutter generated have been well characterised and guidelines for interpretation are in place. However, further artefacts thought to be Taq enzyme slippage leading to products with one repeat unit greater than the parent sequence (n + 4 or forward stutter) or two repeats less (n ? 8 or double back stutter) also occur, but these have not been well characterised despite their potential influence in mixture interpretations. Here we present findings with respect to these additional artefacts from a study of 10,000 alleles and include guidelines for interpretation.     

The spatial and temporal distribution of pollen in a room: Forensic implications

This paper presents two experimental studies that deal with the spatial and temporal distribution of pollen grains within a room of a domestic dwelling. The findings concur with the preliminary work of Morgan et al. [1] and provide greater detail as to the behaviour of pollen grains within indoor locations that are pertinent for forensic investigations. The spatial distribution of pollen in a room exhibits strong distance decay trends, with the majority of pollen recovered within 0.8 m of its source. The pollen was found to persist in increasing quantities during the time the flowers were in the room. This study also shows that 20 days after the flowers were removed, 25–32% of the original pollen was still present within the room. The influence of disturbance was investigated and whilst areas of high disturbance were found to retain less pollen than undisturbed locations, the influence of the proximity to source was a more dominant factor.These findings have significant implications for forensic investigation protocols, particularly the collection and interpretation phases of trace evidence analysis. The distribution of pollen around a room ensures that viable sources of trace pollen are available for transfer if contact is made between a location in the room and a suspect. The persistence of pollen many days after the flowers have been removed from a room indicates that many rooms in domestic dwellings will have distinctive assemblages that reflect the history of the flowers that have been displayed within that room in the past, and that these assemblages will persist and therefore be available for transfer. These preliminary findings indicate that investigation by forensic palynology in indoor domestic settings may well be an underutilised technique that has the potential to provide accurate and valuable intelligence and evidence for forensic enquiry.     

A fatal poisoning involving Bromo-Dragonfly

This paper reports a fatal overdose case involving the potent hallucinogenic drug Bromo-Dragonfly (1-(8-bromobenzo[1,2-b; 4,5-b′]difuran-4-yl)-2-aminopropane). In the present case, an 18-year-old woman was found dead after ingestion of a hallucinogenic liquid. A medico-legal autopsy was performed on the deceased, during which liver, blood, urine and vitreous humour were submitted for toxicological examination. Bromo-Dragonfly was identified in the liver blood using UPLC–TOFMS, and was subsequently quantified in femoral blood (0.0047 mg/kg), urine (0.033 mg/kg) and vitreous humour (0.0005 mg/kg) using LC–MS/MS. Calibration standards were prepared from Bromo-Dragonfly isolated from a bottle found next to the deceased. The structure and purity of the isolated compound were unambiguously determined from analysis of UPLC–TOFMS, GC–MS, HPLC–DAD, ¹H and ¹³C NMR data and by comparison to literature data.The autopsy findings were non-specific for acute poisoning. However, based on the toxicological findings, the cause of death was determined to be a fatal overdose of Bromo-Dragonfly, as no ethanol and no therapeutics or other drugs of abuse besides Bromo-Dragonfly were detected in the liver, blood or urine samples from the deceased. To our knowledge, this is the first report of quantification of Bromo-Dragonfly in a biological specimen from a deceased person. This case caused the drug to be classified as an illegal drug in Denmark on 5th December 2007.     

Analysis of hair after contamination with blood containing cocaine and blood containing benzoylecgonine

In post-mortem work, blood is a potential source of external contamination of hair. The present study was carried out to investigate the amount of drug absorbed into hair which has been contaminated with blood containing either cocaine or BE. Solutions were prepared containing 0.05, 0.1, 0.2, 0.5 and 3.0 μg/mL of either cocaine or BE in human blood. Samples of approximately 3.2 g of drug-free hair were contaminated by soaking in the blood solutions for 5 min. They were then removed and left at room temperature. Approximately 0.5 g of hair was collected from each of the blood soaked hair samples at 6 h, 1, 2, 4 and 7 days after contamination. As each hair sample was collected it was shampoo-washed to prevent further drug absorption. Hair samples were analysed in triplicate using a fully validated method described previously. EME and cocaethylene were also measured in order to find out if cocaine or BE was breaking down to these compounds. Both cocaine and BE were absorbed into hair in significant concentrations when the concentration in the blood was 0.5 μg/mL or greater; cocaine was more readily absorbed than BE. Cocaine broke down to EME (<LOQ) at 0.5 μg/mL and to EME (>LOQ) and BE (<LOQ) at 3.0 μg/mL. When the blood concentration of cocaine was 0.5 μg/mL or less, there was no evidence of it breaking down to form BE. From the samples soaked in blood containing BE, there was no evidence of the BE breaking down. The absorption of drug into hair did not increase as the contamination period increased from 6 h to 7 days.     

Fast quantification of 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid (THCA) using microwave-accelerated derivatisation and gas chromatography–triple quadrupole mass spectrometry

A rapid and sensitive determination of cannabinoids in urine is important in many fields, from workplace drug testing over toxicology to the fight against doping. The detection of cannabis abuse is normally based on the quantification of the most important metabolite 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid (THCA) in urine. In most fields THCA needs to be present at a concentration of exceeding 15 ng/mL before a positive result can be reported.The method described in this paper, combines a 4 min GC–MS/MS method with a fast sample preparation procedure using microwave assisted derivatisation in order to complete the quantification of THCA in urine in 30 min, using only 1 mL of urine.The method is selective, linear over the range 5–100 ng/mL and shows excellent precision and trueness and hence, the estimated measurement uncertainty at the threshold level is small. The method also complies with applicable criteria for mass spectrometry and chromatography. Therefore the method can be used for rapid screening and confirmatory purposes.     

Estimation of legal age using calcification stages of third molars in living individuals

The increased number of adolescents and young adults with unknown or inaccurately given date of birth is a current issue in justice and legal medicine. The objective of this study was to determine the extent to which third molar calcification stages assessed on panoramic X-rays could be useful as additional criteria for forensic age estimation in living individuals, focusing on the legally important ages 17 and 18.In a retrospective multi-center study, the developmental stage of each individual's third molar was analyzed using Demirjian's scale in 2360 cases. Additionally, sex, age and ancestry were assessed.Individuals with the lowest calcification stage of all present molars in stage H were ≥ 18 years with a likelihood of ≥ 99.05% in the female (n = 388), and ≥ 99.24% in the male (n = 482) population.The lowest calcification stage of all present third molars proved to be useful as an additional reliable criterion for the determination of an age ≥ 18 years.     

Relationship between blood and urine alcohol concentrations in apprehended drivers who claimed consumption of alcohol after driving with and without supporting evidence

For various reasons, many people suspected of driving under the influence of alcohol (DUIA) are not apprehended sitting behind the wheel, but some time after the driving. This gives them the opportunity to claim they drank alcohol after the time of driving or after they were involved in a road-traffic crash. Alleged post-offence drinking is not easy for the prosecution to disprove, which often means that the DUIA charge is dropped or the person is acquitted if the case goes to trial. The routine practice of sampling and measuring the concentration of alcohol in blood (BAC) and urine (UAC) and calculating urine/blood ratios (UAC/BAC) and the changes in UAC between two successive voids furnishes useful information to support or challenge alleged drinking after driving. We present here a retrospective case series of DUIA offenders (N = 40) in half of which there was supporting evidence of an after-drink (eye witness or police reports) and in the other half no such evidence existed apart from the suspect's admission. When there was supporting evidence of an after-drink, the UAC/BAC ratio for the first void was close to or less than unity (mean 1.04, median 1.08, range 0.54–1.21) and the UAC increased by 0.21 g/L (range 0.02–0.57) between the two voids. Without any supporting evidence of post-offence drinking the mean UAC/BAC ratio was 1.46 (range 1.35–1.93) for the first void, verifying that absorption and distribution of alcohol in all body fluids and tissues was complete. In these cases, the UAC between successive voids decreased by 0.25 g/L on average (range 0.10–0.49), indicating the post-absorptive phase of the BAC curve. Long experience from investigating claims of post-offence drinking leads us to conclude that in the vast majority of cases this lacks any substance and is simply a last resort by DUIA offenders to evade justice. Unless supporting evidence exists (eye witness, police reports, etc.) of post-offence drinking the courts are encouraged to ignore this defence argument.     

Evaluation of the composition of street cocaine seized in two regions of Brazil

This work evaluates cocaine purity and the concentration ranges of adulterants and inorganic constituents for 31 street cocaine samples seized in two different regions of Brazil from July 2008 to May 2010. Cocaine and adulterants, such as caffeine, lidocaine and benzocaine, were quantified by Gas chromatography–mass spectrometry (GC–MS), and the inorganic constituents were determined by Inductively Coupled Plasma-Optical Emission Spectrometry (ICP-OES) and ion chromatography (IC). The cocaine concentrations in the samples seized in the Amazonas state (AM samples) ranged from 154 to 978 mg g^? 1, and these samples did not contain any of the adulterants studied. The cocaine concentrations in the samples seized in the Minas Gerais state (MG samples) ranged from 63.9 to 753 mg g^? 1. Caffeine was the main adulterant found in 76% of the MG samples, ranging in concentration from 5.5 to 645.3 mg g^? 1. Lidocaine was found in 66.7% of the MG samples, with concentrations ranging from 16.3 to 576.7 mg g^? 1. Benzocaine was found in only one MG sample, at a concentration of 84.8 mg g^? 1. Fourteen elements were identified by ICP-OES, and a wide variation was observed in the concentrations of Ca, Mg, Na, P, Al, Fe, Mn and Zn. Pearson Product–moment Correlations between the analytes allowed the constituents to be associated with the chemicals used in the manufacturing of cocaine and with some common diluents. The study of the purity of cocaine and the presence and concentration of adulterants and inorganic constituents is important because the latter can have deleterious effects on health.     

Detection of acute fentanyl exposure in fresh and decomposed skeletal tissues part II: The effect of dose–death interval

The effects of dose–death interval on the detection of acute fentanyl exposure in fresh and decomposed skeletal tissues (marrow and bone), by automated enzyme-linked immunosorbent assay (ELISA) are described. Rats (n = 14) were administered fentanyl acutely at a dose of 0 (n = 2) or 60 μg/kg (n = 12) by intraperitoneal injection, and euthanized within 20, 45, 135, or 225 min. Femora and tibiae were extracted from the fresh corpses and marrow was isolated from the femoral and tibial medullary cavities. The remains were then allowed to decompose outdoors to the point of complete skeletonization, and vertebrae, pelvi and miscellaneous (humeri and scapulae) were recovered for analysis. In all cases, bones were cleaned in alkaline solution and then ground into a fine powder. Marrow was homogenized in alkaline solution. Fentanyl was extracted from ground bone by methanolic extraction. Extracts were adjusted to pH 6 and analyzed by ELISA. Perimortem heart blood was also collected and diluted in phosphate buffer prior to screening by ELISA. The effect of tissue type on ELISA response was examined through determination of binary classification test sensitivity and the relative decrease in absorbance (%DA, drug-positive tissues vs. drug-free controls) in each tissue type. Overall, the %DA varied significantly between extracts from different skeletal tissues at a given dose–death interval, according to the general order of marrow > decomposed bone > fresh bone. Binary classification test sensitivity values for fentanyl in marrow, fresh epiphyseal (femoral and tibial) bone, fresh diaphyseal (femoral and tibial) bone, decomposed vertebrae, decomposed pelvic bone, and decomposed miscellaneous bone were 67–100%, 0–33%, 0–33%, 0–67%, 0–67% and 0–33%, respectively, over all dose–death intervals. Although group mean %DA values showed a strong negative correlation with dose–death interval in marrow, fresh epiphyseal bone, decomposed vertebrae, pelvic and miscellaneous bone (r = ?0.989, ?0.930, ?0.955, ?0.903, and ?0.974, respectively), the high variability in both fresh and decomposed bone precluded differentiation of the dose–death intervals based on %DA value alone. Overall, the results suggested that the type of skeletal tissue sampled may not be as important as the amount of residual marrow remaining in skeletonized remains.     

The acid phosphatase test two minute cut-off: An insufficient time to detect some semen stains

The ability to detect semen in sexual offence cases is a crucial first step to locating stains which may be suitable for DNA profiling. Since the development of the acid phosphatase test in the late1950s by Stuart S. Kind, the process undertaken to perform the test has gone largely unchanged. The method currently accepted by operational forensic science laboratories allows 2 min for a reaction to be obtained, and until relatively recently, this has not been challenged. In this research, samples of semen were obtained from three donors and a range of dilutions for each sample were prepared. Each dilution was subjected to acid phosphatase testing using both direct testing and the ‘press test’ method. The results showed that semen could be detected in excess of 15 min in dilutions up to 1 in 400 using the press test method and in dilutions up to 1 in 1000 using the direct method. Of further significance was the observation that using the press test method, the two minute cut-off was insufficient to detect the majority of stains and in some cases, semen stains as strong as 1 in 20 dilutions. This research provides compelling evidence for protocols currently utilised in forensic practice to be reviewed in order that forensic scientists do not overlook potential evidential material that may prove suitable for body fluid identification such as DNA STR profiling.     

Potential forensic application of closely linked autosomal STR haplotype in complex kinship testing

It has been noticed that the most commonly used commercial STR kits and mtDNA may not be able to solve some special kinship cases, such as alleged aunt, uncle, niece, nephew or half-siblings. Due to its unique hereditary pattern, the haplotype of genetic markers could be a solution of these questioned family relationships. In this study, we investigated the genetic features of an autosomal STR cluster by employing confirmed family samples. To evaluate the forensic practical value of autosomal STR haplotype, 5 closely linked STR loci, D1S2127-D1S2138-D1S3460-D1S1643-D1S518, which were arranged in about 2 cM region (from 186.29 cM to 188.02 cM; 1 cM represents 1% average recombination between two loci) on chromosome one, were selected to compose haplotype. Genotyping of 60 samples from 8 trios (father–mother–children), 8 duos (father or mother–children), and 4 three-generation pedigrees were performed using PAGE. Haplotypes were identified in the child by determining alleles for all 5 loci transmitted from each parent. Total 73 haplotypes were detected in all samples and 34 haplotypes were observed to be passed down as a whole and was corresponding with the inherited characteristics of haplotype. In all family members, 34 unrelated individuals contributed 65 haplotypes, of which 62 haplotypes appeared only once and the rest 3 haplotypes appeared twice. No recombination was observed in 4 three-generation pedigrees. In conclusion, the haplotype consisting of 5 closely linked autosomal STRs could pass down steadily as a whole. The family specificity of most haplotypes may provide a unique advantage in forensic complex kinship testing.     

Freezing skeletal muscle tissue does not affect its decomposition in soil: Evidence from temporal changes in tissue mass,microbial activity and soil chemistry based on excised samples

The study of decaying organisms and death assemblages is referred to as forensic taphonomy, or more simply the study of graves. This field is dominated by the fields of entomology, anthropology and archaeology. Forensic taphonomy also includes the study of the ecology and chemistry of the burial environment. Studies in forensic taphonomy often require the use of analogues for human cadavers or their component parts. These might include animal cadavers or skeletal muscle tissue. However, sufficient supplies of cadavers or analogues may require periodic freezing of test material prior to experimental inhumation in the soil. This study was carried out to ascertain the effect of freezing on skeletal muscle tissue prior to inhumation and decomposition in a soil environment under controlled laboratory conditions. Changes in soil chemistry were also measured. In order to test the impact of freezing, skeletal muscle tissue (Sus scrofa) was frozen (?20 °C) or refrigerated (4 °C). Portions of skeletal muscle tissue (～1.5 g) were interred in microcosms (72 mm diameter × 120 mm height) containing sieved (2 mm) soil (sand) adjusted to 50% water holding capacity. The experiment had three treatments: control with no skeletal muscle tissue, microcosms containing frozen skeletal muscle tissue and those containing refrigerated tissue. The microcosms were destructively harvested at sequential periods of 2, 4, 6, 8, 12, 16, 23, 30 and 37 days after interment of skeletal muscle tissue. These harvests were replicated 6 times for each treatment. Microbial activity (carbon dioxide respiration) was monitored throughout the experiment. At harvest the skeletal muscle tissue was removed and the detritosphere soil was sampled for chemical analysis. Freezing was found to have no significant impact on decomposition or soil chemistry compared to unfrozen samples in the current study using skeletal muscle tissue. However, the interment of skeletal muscle tissue had a significant impact on the microbial activity (carbon dioxide respiration) and chemistry of the surrounding soil including: pH, electroconductivity, ammonium, nitrate, phosphate and potassium. This is the first laboratory controlled study to measure changes in inorganic chemistry in soil associated with the decomposition of skeletal muscle tissue in combination with microbial activity.     

An investigation into the effects of decomposition on the post-mortem location of trans-dermal artefacts

A search area of a crime scene to recover trans-dermal artefacts (here, earrings) was conducted using 12 pig carcasses, of various sizes, with pierced ears; 6 were buried and 6 deposited on the surface in a wooded area. After 28 months, the remains were excavated and recovered, and the final location of the earrings recorded. The furthest recorded earring from its associated surface remains was 119 cm. In the buried remains, on three occasions earrings were found located 6 cm below the recorded base of the grave. Formal recommendations for the search area of a crime scene have been established as 120 cm radius from the originally located remains, whether surface deposition or burial, and confirmation of excavation below the assumed floor of the grave in burials is essential, at least to 10 cm.     

Experimental forensic studies of the preservation of pollen in vehicle fires

The implications of the recent recommendations of the Law Commission regarding the use of admissibility tests have the potential to be far reaching for forensic disciplines that rely on the expertise of highly qualified expert witnesses. These disciplines will need a concomitant body of peer-reviewed experiments that provides a basis for the interpretations of such evidence presented in court. This paper therefore, presents such results from two experiments which were undertaken to address specific issues that were raised in cases presented in the British courtroom. These studies demonstrate that there is a variability in the persistence of Lily, Daffodil and Tulip pollen when exposed to high temperatures between 0.5 min and 1440 min (24 h). It was possible to identify all three pollen types after 30 min of exposure to 400 °C, and after shorter time frames the threshold for successful identification was 700 °C after 0.5 min for all pollen types tested and 500 °C for Daffodil and Lily after 5 min of heat exposure. Over longer time periods (18 h (1080 min)) the different pollen types were found to persist in a viable form for identification at 300 °C (Lily), 200 °C (Daffodil) and 50 °C (Tulip). These findings, albeit from a small sample of pollen types, provide empirical contextual information that would contribute to such evidence having sufficient scientific weight to meet admissibility criteria and be viable evidence for a court. These studies demonstrate the value in seeking pollen evidence from even such extreme crime scenes as encountered in vehicular fires.