Characteristics of Forensic Imaging Performance—An Analysis of Forensic Imaging Bottlenecks

Disk imaging involves copying all of the data from a source disk drive to a target. Typically, the target for the copy is another disk drive. Forensic processes developed years ago do not appear to be adequate for current storage technology. For example, with disk drive capacities now exceeding 1 Terabyte, a typical disk imaging can take over 8 hours at typical rates. With disk drive capacities increasing, forensic copying is expected to take even longer. Along with increase in disk capacity, the industry has also seen an increase in data transfer rates. In many cases, forensic imaging is taking longer than necessary. To identify the bottlenecks, an examination of different methods used to transfer data from a source disk was performed. Factors considered were differing disk access technologies. One finding is that the USB disk access technology (version 2.0 and earlier) is a significant bottleneck for data transfer rates, especially when the USB device is a write‐blocker. Other factors that contribute to the efficiency of a forensic copy are the file system used to write a forensic image and the data transfer size used when reading from a disk drive. Optimal parameters for performing a forensic acquisition from a disk drive are identified.     

Investigations into Enhancing Yersinia pestis Cells Viability following Environmental Sampling for Forensic Analysis

Following an intentional or accidental bio-warfare agent (BWA) release, environmental sample analysis is absolutely critical to determine the extent of contamination. When dealing with nonspore forming BWA (e.g., Yersinia pestis), retention of cell viability is central to such analyses. Even though significant advances have been achieved in DNA sequencing technologies, a positive identification of BWAs in environmental samples must be made through the ability of cells to form colony-forming units upon culturing. Inability to revive the cells between collection and analysis renders such studies inconclusive. Commercial kits designed to preserve the viability of pathogens contained within clinical samples are available, but many of them have not been examined for their ability to preserve samples containing suspected BWAs. The study was initiated to examine the applicability of commercial solutions aiding in retention of Y. pestis viability in samples stored under nonpermissive temperatures, that is, 40 and 37°C. While none of the tested solutions sustained cell viability at 40°C, the results show five out of 17 tested preservatives were capable of supporting viability of Y. pestis at 37°C.     

Detection of Tooth‐Colored Restorative Materials for Forensic Purposes Based on Their Optical Properties: An In Vitro Comparative Study

Victim identification using dental records involves antemortem and postmortem comparison of dental charts. Since dental restorations may be part of such records, identifying them accurately is critical. The objective of this study was to compare the diagnostic reliability and validity of two optical methods for identifying tooth‐colored restorations (digital imaging fiber optic transillumination (DiFOTI) using near infrared light, and fluorescence‐aided identification of restorations (FAIR)) with conventional diagnostic methods. Four examiners identified and charted tooth‐colored restorations in three sets of typodonts on the bench using conventional visual and tactile examination, DiFOTI (DIAGNOcam?) and FAIR. All examinations were repeated after 4 weeks. Both the sensitivity (95%) and specificity (97%) of the FAIR method were significantly higher than those for DiFOTI (82% and 82%) and for conventional inspection (71% and 82%). In conclusion, FAIR method performed better than conventional examination and DiFOTI, and was more reliable for identifying tooth‐colored restorations.     

A Comparative Taphonomic Analysis of 24 Trophy Skulls from Modern Forensic Cases,

Cranial remains retained from fallen enemies are commonly referred to as “trophy skulls,” and many such crania were acquired as souvenirs by U.S. servicemembers during WWII and the Vietnam conflict. These remains increasingly have become the subject of forensic anthropological analysis as their possessors, typically veterans or their relatives, try to discard or repatriate them. The present research uses a qualitative analytical approach to review 24 cases of reported trophy skulls (14 previously unpublished cases and 10 from the literature) to determine which perimortem and postmortem characteristics are most useful for generating a taphonomic profile. Overall, the taphonomic signature of trophy remains includes traits relating to acquisition and preparation, ornamental display, and subsequent curation. Contextual evidence and the biological profile also are considered when determining the possible origin of human cranial remains as a trophy skull. Thorough taphonomic analysis will aid in identifying these types of remains as trophy skulls.     

Ultrafast STR Separations on Short‐Channel Microfluidic Systems for Forensic Screening and Genotyping

There are situations in which it is important to quickly and positively identify an individual. Examples include suspects detained in the neighborhood of a bombing or terrorist incident, individuals detained attempting to enter or leave the country, and victims of mass disasters. Systems utilized for these purposes must be fast, portable, and easy to maintain. DNA typing methods provide the best biometric information yielding identity, kinship, and geographical origin, but they are not portable and rapid. This study details the development of a portable short‐channel microfluidic device based on a modified Agilent 2100 bioanalyzer for applications in forensic genomics. The system utilizes a denaturing polymer matrix with dual‐channel laser‐induced fluorescence and is capable of producing a genotype in 80 sec. The device was tested for precision and resolution using an allelic ladder created from 6 short tandem repeat (STR) loci and a sex marker (amelogenin). The results demonstrated a precision of 0.09–0.21 bp over the entire size range and resolution values from 2.5 to 4.1 bp. Overall, the results demonstrate the chip provides a portable, rapid, and precise method for screening amplified short tandem repeats and human identification screening.     

High‐Resolution Melting (HRM) of Hypervariable Mitochondrial DNA Regions for Forensic Science

Forensic strategies commonly are proceeding by analysis of short tandem repeats (STRs); however, new additional strategies have been proposed for forensic science. Thus, this article standardized the high‐resolution melting (HRM) of DNA for forensic analyzes. For HRM, mitochondrial DNA (mtDNA) from eight individuals were extracted from mucosa swabs by DNAzol reagent, samples were amplified by PCR and submitted to HRM analysis to identify differences in hypervariable (HV) regions I and II. To confirm HRM, all PCR products were DNA sequencing. The data suggest that is possible discriminate DNA from different samples by HRM curves. Also, uncommon dual‐dissociation was identified in a single PCR product, increasing HRM analyzes by evaluation of melting peaks. Thus, HRM is accurate and useful to screening small differences in HVI and HVII regions from mtDNA and increase the efficiency of laboratory routines based on forensic genetics.     

Hydrocarbon and Fatty Acid Composition from Blowfly Eggs Represents a Potential Complementary Taxonomic Tool of Forensic Importance,

One of the most important contributions of forensic entomology is to assist criminal expertise to determine the postmortem interval, which depends on the duration of the immature stages of insects of forensic interest. On the other hand, the time of development of the different stages varies according to the species; therefore, its identification is essential. Currently, few studies have investigated the use of cuticular hydrocarbons, and none regarding fatty acids, as complementary taxonomic tools to expedite species identification. Therefore, we evaluated whether cuticular hydrocarbons together with fatty acids of eggs of flies of the family Calliphoridae, main group of forensic interest, can be used to distinguish species. The analyses were performed by chromatographic techniques. The results show that there are significant differences between the composition of cuticular hydrocarbons and fatty acids between species and, therefore, they can be used to provide a complementary taxonomic tool to expedite the forensic expertise.     

Detection of Anthrax and Other Pathogens Using a Unique Liquid Array Technology

A bead‐based liquid hybridization assay, Luminex^® 100?, was used to identify four pathogenic bacteria, Bacillus anthracis, Clostridium botulinum, Francisella tularensis subsp. tularensis, and Yersinia pestis, and several close relatives. Hybridization between PCR‐amplified target sequences and probe sequences (located within the 23S ribosomal RNA gene rrl and the genes related to the toxicity of each bacterium) was detected in single‐probe or multiple‐probe assays, depending on the organism. The lower limits of detection (LLDs) for the probes ranged from 0.1 to 10 ng. Sensitivity was improved using lambda exonuclease to digest the noncomplementary target strand. All contributors in 33 binary, ternary, and quaternary mixtures in which all components were present in a 1:1 ratio were identified with an 80% success rate. Twenty‐eight binary mixtures in which the two components were combined in various ratios were further studied. All target sequences were detected, even when the minor component was overshadowed by a tenfold excess of the major component.     

Forensic Use of the Piracatinga Fish (Callophysius macropterus) to Locate and Identify Human Remains Retrieved From the Amazon River

Piracatinga (Callophysius macropterus) are a type of bottom feeder catfish encountered in the Amazon River and its tributaries. We report two cases in which human remains were first located based on a characteristic circular distortion of the surface of the river that the Piracatinga make while they feed. Human skin samples of one of the victims recovered from the Piracatinga digestive tract were subjected to mitochondrial DNA analysis that allowed identification of the body of Case 1; the family recognized body parts of Case 2. Importantly, the location of human body parts and their identification based on DNA analysis enabled the respective families to obtain a death certificate expeditiously in the absence of identifiable remains—a process that normally requires 5 years under current Brazilian law, and in the absence of closure, imposes severe emotional stress on the family of the deceased.     

The Effect of Flunitrazepam (Rohypnol®) on the Development of Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae) and its Implications for Forensic Entomology

This study investigated the potential effects of flunitrazepam (known as “date rape drug”) on the developmental cycle of Chrysomya megacephala, an important forensic species, and their possible implications for the calculation of the PMI. A 1050 C. megacephala eggs were divided into five groups with seven replications each. The eggs were placed on artificial diet prepared with four drug concentrations of flunitrazepam (4, 8, 16, and 32 ng/g), besides the control group (prepared with water). Were evaluated the potential effects on development time, weight gain, and mortality during the cycles. The drug had no significant effect on development time or mortality although it did affect the weight of the pupae and adults (Kruskal–Wallis, p < 0.05). The result can be deduced that the determination of the postmortem interval is not affected.     

The Impact of Daubert on the Admissibility of Forensic Anthropology Expert Testimony

Forensic anthropologists anticipated a significant impact from the 1993 Supreme Court Daubert decision, which addressed the standard of admissibility for expert testimony. In response, many forensic articles cited Daubert in the search for objective techniques or a critique of established subjective methods. This study examines challenges to forensic anthropological expert testimony to evaluate whether Daubert has actually affected the admissibility of such testimony. Thirty cases were identified that addressed the admissibility of the testimony, including 14 cases prior to Daubert and 16 after Daubert. Examination of these cases indicates that post‐Daubert cases do not result in more exclusions. Yet, this lack of exclusions may instead be viewed as a manifestation of the field's overall surge toward more objective and quantifiable techniques in a self‐regulating response to Daubert.     

An Optimized Centrifugal Method for Separation of Semen from Superabsorbent Polymers for Forensic Analysis

Connection of a perpetrator to a sexual assault is best performed through the confirmed presence of semen, thereby proving sexual contact. Evidentiary items can include sanitary napkins or diapers containing superabsorbent polymers (SAPs), complicating spermatozoa visualization and DNA analysis. In this report, we evaluated the impact of SAPS on the current forensic DNA workflow, developing an efficient centrifugal protocol for separating spermatozoa from SAP material. The optimized filtration method was compared to common practices of excising the top layer only, resulting in significantly higher sperm yields when a core sample of the substrate was taken. Direct isolation of the SAP‐containing materials without filtering resulted in 20% sample failure; additionally, SAP material was observed in the final eluted DNA samples, causing physical interference. Thus, use of the described centrifugal‐filtering method is a simple preliminary step that improves spermatozoa visualization and enables more consistent DNA yields, while also avoiding SAP interference.     

The Use of Forensic Tests to Distinguish Blowfly Artifacts from Human Blood,Semen, and Saliva

This study investigated whether routinely used forensic tests can distinguish 3‐day‐old or 2‐week‐old fly artifacts, produced after feeding on human blood, semen, or saliva, from the biological fluid. Hemastix^®, Hemident^?, and Hemascein^? were unable to distinguish blood from artifacts. Hemastix^® returned false positives from negative controls. ABAcard^® Hematrace^® and Hexagon OBTI could distinguish blood from 3‐day‐old artifacts, but not 2‐week‐old artifacts. Phadebas^® and SALIgAE^® were unable to distinguish saliva from artifacts. RSID^?‐Saliva was able to distinguish saliva from 3‐day‐old artifacts, but not 2‐week‐old artifacts. Semen tests Seminal Acid Phosphatase, RSID^?‐Semen, and ABAcard^® p30 were all able to distinguish semen from 3‐day‐old artifacts, but not 2‐week‐old artifacts. The tests investigated cannot be relied upon to distinguish artifacts from biological fluids. However, if an artifact is identified by its morphology, a positive result may indicate which biological fluid the fly consumed, and this knowledge may prove useful for investigators searching for DNA.     

Bacterial Cross‐contamination Potential Associated with Contaminated Currency

Previous studies have shown a significant amount of contaminants on paper currencies. It is important to study the transfer of microorganisms between paper currencies to determine whether it meets the level of a human health threat. This cross‐contamination potential was analyzed by seeding new US 1‐dollar bills with Bacillus thuringiensis, and pressing or rubbing them against clean currency to determine the amount of bacteria transfer to the unseeded currency. The transferred amount of bacteria was recovered, plated, incubated, and the colony‐forming units were quantified. Among the recovery methods tested, the most efficient method, vortexing for 10 min with a recovery efficiency of 40 ± 8.1%, was used in this analysis. The resulting transfer rates were 4.8%, 8.6%, and 14.3% when pressed for 24 h, 72 h, and rubbed together, respectively. These transferred amounts of bacteria are significant and have the potential to spread infectious diseases.     

Rapid Quantitative Analysis of Multiple Explosive Compound Classes on a Single Instrument via Flow‐Injection Analysis Tandem Mass Spectrometry

A flow‐injection analysis tandem mass spectrometry (FIA MSMS) method was developed for rapid quantitative analysis of 10 different inorganic and organic explosives. Performance is optimized by tailoring the ionization method (APCI/ESI), de‐clustering potentials, and collision energies for each specific analyte. In doing so, a single instrument can be used to detect urea nitrate, potassium chlorate, 2,4,6‐trinitrotoluene, 2,4,6‐trinitrophenylmethylnitramine, triacetone triperoxide, hexamethylene triperoxide diamine, pentaerythritol tetranitrate, 1,3,5‐trinitroperhydro‐1,3,5‐triazine, nitroglycerin, and octohy‐dro‐1,3,5,7‐tetranitro‐1,3,5,7‐tetrazocine with sensitivities all in the picogram per milliliter range. In conclusion, FIA APCI/ESI MSMS is a fast (<1 min/sample), sensitive (~pg/mL LOQ), and precise (intraday RSD < 10%) method for trace explosive detection that can play an important role in criminal and attributional forensics, counterterrorism, and environmental protection areas, and has the potential to augment or replace several of the existing explosive detection methods.     

Effect of Heat on the Fluorescence Properties of Tooth‐Colored Restorative Materials and Their Forensic Implications

During antemortem and postmortem comparison of dental records of carbonized victims, restorations may be part of such records. The objective of this study was to evaluate the effect of heat on the fluorescence behavior of contemporary tooth‐colored restorative materials and natural tooth structure when subjected to range of temperatures, using illumination with 405 nm wavelength light. A total of 132 human extracted teeth restored with tooth‐colored restorative materials were exposed to heat (200, 500, 900, 1200°C) in an oven for 30 min. Samples were imaged before and after heat treatment. All tooth‐colored restorative materials underwent changes in color and in fluorescence properties, at each of the temperatures used. Resin‐based restorative materials still fluoresced at 200°C, and at 500°C underwent major color changes due to volatilization of resin. Materials containing inorganic fluorophores still fluoresced at 900°C, while at 1200°C, none of the materials tested in this study showed any fluorescence.     

A GIS‐based Quantitative Approach for the Search of Clandestine Graves,Italy

Previous research on the RAG color‐coded prioritization systems for the discovery of clandestine graves has not considered all the factors influencing the burial site choice within a GIS project. The goal of this technical note was to discuss a GIS‐based quantitative approach for the search of clandestine graves. The method is based on cross‐referenced RAG maps with cumulative suitability factors to host a burial, leading to the editing of different search scenarios for ground searches showing high‐(Red), medium‐(Amber), and low‐(Green) priority areas. The application of this procedure allowed several outcomes to be determined: If the concealment occurs at night, then the “search scenario without the visibility” will be the most effective one; if the concealment occurs in daylight, then the “search scenario with the DSM‐based visibility” will be most appropriate; the different search scenarios may be cross‐referenced with offender's confessions and eyewitnesses’ testimonies to verify the veracity of their statements.     

In‐Depth Analysis of Computer Memory Acquisition Software for Forensic Purposes

The comparison studies on random access memory (RAM) acquisition tools are either limited in metrics or the selected tools were designed to be executed in older operating systems. Therefore, this study evaluates widely used seven shareware or freeware/open source RAM acquisition forensic tools that are compatible to work with the latest 64‐bit Windows operating systems. These tools' user interface capabilities, platform limitations, reporting capabilities, total execution time, shared and proprietary DLLs, modified registry keys, and invoked files during processing were compared. We observed that Windows Memory Reader and Belkasoft's Live Ram Capturer leaves the least fingerprints in memory when loaded. On the other hand, ProDiscover and FTK Imager perform poor in memory usage, processing time, DLL usage, and not‐wanted artifacts introduced to the system. While Belkasoft's Live Ram Capturer is the fastest to obtain an image of the memory, Pro Discover takes the longest time to do the same job.     

SNP Miniplexes for Individual Identification of Random‐Bred Domestic Cats

Phenotypic and genotypic characteristics of the cat can be obtained from single nucleotide polymorphisms (SNPs) analyses of fur. This study developed miniplexes using SNPs with high discriminating power for random‐bred domestic cats, focusing on individual and phenotypic identification. Seventy‐eight SNPs were investigated using a multiplex PCR followed by a fluorescently labeled single base extension (SBE) technique (SNaPshot^®). The SNP miniplexes were evaluated for reliability, reproducibility, sensitivity, species specificity, detection limitations, and assignment accuracy. Six SNPplexes were developed containing 39 intergenic SNPs and 26 phenotypic SNPs, including a sex identification marker, ZFXY. The combined random match probability (cRMP) was 6.58 × 10^?19 across all Western cat populations and the likelihood ratio was 1.52 × 10¹⁸. These SNPplexes can distinguish individual cats and their phenotypic traits, which could provide insight into crime reconstructions. A SNP database of 237 cats from 13 worldwide populations is now available for forensic applications.     

Genetic Identification of Communist Crimes’ Victims (1944–1956) Based on the Analysis of One of Many Mass Graves Discovered on the Powazki Military Cemetery in Warsaw,Poland

As the result of the communist terror in Poland, during years 1944–1956 more than 50,000 people died. Their bodies were buried secretly, and most places are still unknown. The research presents the results of identification of people buried in one of many mass graves, which were found at the cemetery Pow?zki Military in Warsaw, Poland. Exhumation revealed the remains of eight people, among which seven were identified genetically. Well‐preserved molars were used for the study. Reference material was collected from the closest living relatives. In one case, an exhumation of victim's parents had to be performed. DNA from swabs was extracted with a PrepFiler^® BTA Forensic DNA Extraction Kit and organic method. Autosomal, Y‐STR amplification, and mtDNA sequencing were performed. The biostatistical calculations resulted in LR values from 1608 to 928 × 10¹⁸. So far, remains of more than 50 victims were identified.