Search

Compositional data from a soil survey over North Canberra, Australian Capital Territory, are used to develop and test an empirical soil provenancing method. Mineralogical data from Fourier transform infrared spectroscopy (FTIR) and magnetic susceptibility (MS), and geochemical data from X‐ray fluorescence (XRF; for total major oxides) and inductively coupled plasma‐mass spectrometry (ICP‐MS; for both total and aqua regia‐soluble trace elements) are performed on the survey's 268 topsoil samples (0–5 cm depth; 1 sample per km2). Principal components (PCs) are calculated after imputation of censored data and centered log‐ratio transformation. The sequential provenancing approach is underpinned by (i) the preparation of interpolated raster grids of the soil properties (including PCs); (ii) the explicit quantification and propagation of uncertainty; (iii) the intersection of the soil property rasters with the values of the evidentiary sample (± uncertainty); and (iv) the computation of cumulative provenance rasters (“heat maps”) for the various analytical techniques. The sequential provenancing method is tested on the North Canberra soil survey with three “blind” samples representing simulated evidentiary samples. Performance metrics of precision and accuracy indicate that the FTIR and MS (mineralogy), as well as XRF and total ICP‐MS (geochemistry) analytical methods, offer the most precise and accurate provenance predictions. Inclusion of PCs in provenancing adds marginally to the performance. Maximizing the number of analytes/analytical techniques is advantageous in soil provenancing. Despite acknowledged limitations and gaps, it is concluded that the empirical soil provenancing approach can play an important role in forensic and intelligence applications.

In this study, we aim to compare the performance of systems and forensic facial comparison experts in terms of likelihood ratio computation to assess the potential of the machine to support the human expert in the courtroom. In forensics, transparency in the methods is essential. Consequently, state‐of‐the‐art free software was preferred over commercial software. Three different open‐source automated systems chosen for their availability and clarity were as follows: OpenFace, SeetaFace, and FaceNet; all three based on convolutional neural networks that return a distance (OpenFace, FaceNet) or similarity (SeetaFace). The returned distance or similarity is converted to a likelihood ratio using three different distribution fits: parametric fit Weibull distribution, nonparametric fit kernel density estimation, and isotonic regression with pool adjacent violators algorithm. The results show that with low‐quality frontal images, automated systems have better performance to detect nonmatches than investigators: 100% of precision and specificity in confusion matrix against 89% and 86% obtained by investigators, but with good quality images forensic experts have better results. The rank correlation between investigators and software is around 80%. We conclude that the software can assist in reporting officers as it can do faster and more reliable comparisons with full‐frontal images, which can help the forensic expert in casework.

A wet‐vacuum‐based collection method with the M‐Vac® was compared to a wet‐swabbing collection method by examining the recovery of diluted blood on 22 substrates of varying porosity. The wet‐vacuum method yielded more total nuclear DNA than wet‐swabbing on 18 porous substrates, recovering on average 12 times more DNA. However, both methods yielded comparable amounts of total DNA on two porous and two nonporous substrates. In no instance did wet‐swabbing significantly recover more DNA. The wet‐vacuum method also successfully collected additional DNA on previously swabbed substrates. Mitochondrial DNA yields were assessed, and outcomes were generally similar to the nuclear DNA outcomes described above. Results demonstrate that wet‐vacuuming may serve as an alternative collection method to swabbing on difficult porous substrates and could potentially recover additional DNA on previously swabbed substrates. However, swabbing remains the preferred collection method on substrates with visible stains and/or nonporous surfaces for reasons of convenience, simplicity, and lower cost relative to the wet‐vacuum method.

The need for agile and proper identification of drugs of abuse has encouraged the scientific community to improve and todevelop new methodologies. The drug lysergic acid diethylamide (LSD) is still widely used due to its hallucinogenic effects. The use ofvoltammetric methods to analyze narcotics has increased in recent years, and the possibility of miniaturizing the electrochemical equipmentallows these methods to be applied outside the laboratory; for example, in crime scenes. In addition to portability, the search for affordable andsustainable materials for use in electroanalytical research has grown in recent decades. In this context, employing paper substrate, graphite pen-cil, and silver paint to construct paper-based electrodes is a great alternative. Here, a paper-based device comprising three electrodes was drawnon 300 g/m2watercolor paper with 8B pencils, and its efficiency was compared to the efficiency of a commercially available screen-printedcarbon electrode. Square wave voltammetry was used for LSD analysis in aqueous medium containing 0.05 mol/L LiClO4. The limits of detec-tion and quantification were 0.38 and 1.27 mol/L, respectively. Both electrodes exhibited a similar voltammetric response, which was alsoconfirmed during analysis of a seized LSD sample, with recovery of less than 10%. The seized samples were previously analyzed by GCMStechnique, employing the full scan spectra against the software spectral library. The electrode selectivity was also tested against 3,4-methylene-dioxymethamphetamine (MDMA) and methamphetamine. It was possible to differentiate these compounds from LSD, indicating that the devel-oped paper-based device has potential application in forensic chemistry analyses., Financial support provided by the Polícia Científica do Estado de São Paulo for the partnership, and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP - Process 2016/23825-3), Fundação para a Ciência e Tecnologia, FCT, Portugal, (UID/QUI/00686/2016 and UID/QUI/00686/2019), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Process Capes Pro Forenses 25/2014).

Postmortem computed tomography (PMCT) is integrated into the evaluation of decedents in several American medical examiner offices and medicolegal death investigative centers in many other countries. We retrospectively investigated the value of PMCT in a series of firearm homicide cases from a statewide centralized medical examiner’s office that occurred during 2016. Autopsies were performed or supervised by board‐certified forensic pathologists who reviewed the PMCT scans prior to autopsy. PMCT scans were re‐evaluated by a forensic radiologist blinded to the autopsy findings and scored by body region (head–neck, thoracoabdominal, and extremities). Injury discrepancies were scored using a modified Goldman classification and analyzed with McNemar’s test. We included 60 males and 20 females (median age 31 years, range 3–73). Based on PMCT, 56 (79.1%) cases had injuries relevant to the cause of death in a single body region (24 head–neck region, 32 thoracoabdominal region). Out of these 56 cases, 9 had a missed major diagnosis by PMCT outside that region, including 6 extremity injuries visible during standard external examination. Yet all had evident lethal firearm injury. We showed that PMCT identifies major firearm injuries in homicide victims and excludes injuries related to the cause of death in other regions when a single body region is injured. Although PMCT has a known limited sensitivity for soft tissue and vascular pathology, it can be combined with external examination to potentially reduce or focus dissections in some of these cases depending on the circumstances and medicolegal needs.

In November 2018, Butte County, California, was decimated by the Camp Fire, the deadliest wildfire in state history. Over 150,000 acres were destroyed, and at its peak, the fire consumed eighty acres per minute. The speed and intensity of the oncoming flames killed scores of people, and weeks before the fire was contained, first responders began searching through the rubble of 18,804 residences and commercial buildings. As with most mass disasters, conventional identification modalities (e.g., fingerprints, odontology, hardware) were utilized to identify victims. The intensity and duration of the fire severely degraded most of the remains, and these approaches were useful in only 22 of 84 cases. In the past, the remaining cases would have been subjected to conventional DNA analysis, which may have required months to years. Instead, Rapid DNA technology was utilized (in a rented recreational vehicle outside the Sacramento morgue) in the victim identification effort. Sixty‐nine sets of remains were subjected to Rapid DNA Identification and, of these, 62 (89.9%) generated short tandem repeat profiles that were subjected to familial searching; essentially all these profiles were produced within hours of sample receipt. Samples successfully utilized for DNA identification included blood, bone, liver, muscle, soft tissue of unknown origin, and brain. In tandem with processing of 255 family reference samples, 58 victims were identified. This work represents the first use of Rapid DNA Identification in a mass casualty event, and the results support the use of Rapid DNA as an integrated tool with conventional disaster victim identification modalities.

A developmental validation was performed to demonstrate reliability, reproducibility, and robustness of the ANDE Rapid DNA Identification System for processing of crime scene and disaster victim identification (DVI) samples. A total of 1705 samples were evaluated, including blood, oral epithelial samples from drinking containers, samples on FTA and untreated paper, semen, bone, and soft tissues. This study was conducted to address the FBI’s Quality Assurance Standards on developmental validation and to accumulate data from a sufficient number of unique donors and sample types to meet NDIS submission requirements for acceptance of the ANDE Expert System for casework use. To date, no Expert System has been approved for such samples, but the results of this study demonstrated that the automated Expert System performs similarly to conventional laboratory data analysis. Furthermore, Rapid DNA analysis demonstrated accuracy, precision, resolution, concordance, and reproducibility that were comparable to conventional processing along with appropriate species specificity, limit of detection, performance in the presence of inhibitors. No lane‐to‐lane or run‐to‐run contamination was observed, and the system correctly identified the presence of mixtures. Taken together, the ANDE instrument, I‐Chip consumable, FlexPlex chemistry (a 27‐locus STR assay compatible with all widely used global loci, including the CODIS core 20 loci), and automated Expert System successfully processed and interpreted more than 1200 unique samples with over 99.99% concordant CODIS alleles. This extensive developmental validation data provides support for broad use of the system by agencies and accredited forensic laboratories in single‐source suspect‐evidence comparisons, local database searches, and DVI.

Bruises are often difficult to detect on victims of violence, potentially impacting investigation and prosecution. The purpose of our randomized controlled trial was to measure the effectiveness of an alternate light source (ALS) within visible and long ultraviolet spectrums at improving bruise detection compared to white light over time. We also examined the effects of skin color, age, gender, localized fat, and injury mechanism on bruise detection. Participants included 157 healthy adults with balanced sampling across six skin color categories. Bruises were created under the controlled application of a paintball pellet and dropped weight to one upper and lower arm, respectively. Using a crossover design, both bruises were examined 21 times over 4 weeks. Ten different wavelength (350–535 nm) and filter (yellow, orange, red) combinations were used. Multilevel models were used to analyze 2903 examinations on both upper and lower arms. Results in multivariable models showed after controlling for other covariates 415 and 450 nm using a yellow filter had greater odds of detecting evidence of bruising than white light (Upper Arm: 415 nm: OR = 5.34, 95% CI: 4.35–6.56; 450 nm: OR = 4.08, 95% CI: 3.36–4.96). Under either light source, being female and having more localized fat had increased odds of detecting bruises created by the dropped weight (female: OR = 2.96, 95% CI: 2.37–3.70; fat: OR = 1.21, 95% CI: 1.09–1.34). Our results support ALS as an appropriate tool to enhance concurrent physical assessment of bruises in the presence of known history of injury. Future development and evaluation of clinical practice guidelines for ALS application are needed.

In 2007, the Bureau of Justice Statistics reported on 2004 data collected from the Census of Medical Examiner and Coroner Offices (CMEC). The CMEC was one of the first comprehensive reports on the state of the medicolegal death investigation system in the United States and included information on administration, expenditure, workload, specialized death investigations, records and evidence retention, and resources. However, the report did not include responses on questions that were related to toxicology such as specimen retention and type of testing. The purpose of this publication is to provide the community with toxicology laboratory‐specific responses from nearly 2000 medical examiner and coroner (MEC) offices. Data obtained from a BJS CMEC public use dataset for any remaining information that was not reported in the 2007 BJS report were evaluated specific to the operation of toxicology laboratories within a MEC office or specific to toxicology testing. The CMEC includes information on average operating budget for MEC offices with internal or external toxicology services, budget for toxicology/microbiology services, respondents’ routine uses of toxicology analysis, toxicology specimen retention time, average turnaround times, use of computerized information management systems, and participation in federal data collections. These historical data begin to address the present state of our nation’s toxicology laboratories within the medicolegal death investigation system and their preparedness for the current drug overdose epidemic.

This study examined associations between criminal recidivism after discharge from forensic treatment and variables related to either the time before the current forensic treatment, or the current forensic treatment, or the follow‐up after discharge. Participants were treated in 12 forensic clinics according to section 63 of the German penal code. A patient was classified as a criminal recidivist when the patient or the aftercare reported that the patient was delinquent at follow‐up. Patients without criminal recidivism were patients for which both perspectives (patient and aftercare) reported no delinquency at follow‐up. Mann–Whitney U‐tests and Fisher's exact tests were performed. Data to classify patients were available for N = 249 patients. Fifteen patients (6%) were classified as criminal recidivists. The follow‐up was M = 12.58 (SD = 1.84) months, and the criminal acts occurred M = 6.00 (SD = 5.55) months after discharge. Differences between patients with and without criminal recidivism were found in pretreatment (young age at first crime, early onset of mental disorder, previous forensic treatments), treatment‐related (disorder due to psychoactive substance use, gradual release abuses, outbreaks, assaults against staff, criminal act during treatment, type of discharge, outcome ratings), as well as follow‐up variables (no specified housing situation, not being abstinent from psychoactive substances, inpatient readmission, course of outpatient treatment, course of mental disorder) (all p

Improvements have been made in identifying the prevalence of risk factors/indicators for violent extremism. A consistent problem is the lack of base rates. How to develop base rates is of equal concern. This study has two aims: (i) compare two methods for developing base rates; the Unmatched Count Technique (UCT) and direct questioning, (ii) generate base rates in a general population sample and compare these to a sample of lone‐actor terrorists (n = 125). We surveyed 2108 subjects from the general population. Participants were recruited from an online access panel and randomly assigned to one of three conditions; direct survey, control, or UCT. Survey items were based on a lone‐actor terrorist codebook developed from the wider literature. Direct questioning was more suitable under our study conditions where UCT resulted in deflation effects. Comparing the base rates identified a number of significant differences: (i) lone‐actor terrorists demonstrated propensity indicators related to a cognitive susceptibility, and a crime‐ and/or violence‐supportive morality more often; the general sample demonstrated protective factors more often, (ii) lone‐actor terrorists demonstrated situational indicators related to a crime‐ and/or violence‐supportive morality more often, whereas the general sample experienced situational stressors more often, (iii) lone‐actor terrorists demonstrated indicators related to exposure to extremism more often. Results suggest there are measurable differences in the prevalence of risk factors between lone‐actor terrorists and the general population. However, no single factor “predicts” violent extremism. This bears implications for our understanding of the interrelation of risk and protective factors, and for the risk assessment of violent extremism.

Recent reports have demonstrated that genetically variant peptides derived from human hair shaft proteins can be used to differentiate individuals of different biogeographic origins. We report a method involving direct extraction of hair shaft proteins more sensitive than previously published methods regarding GVP detection. It involves one step for protein extraction and was found to provide reproducible results. A detailed proteomic analysis of this data is presented that led to the following four results: (i) A peptide spectral library was created and made available for download. It contains all identified peptides from this work, including GVPs that, when appropriately expanded with diverse hair‐derived peptides, can provide a routine, reliable, and sensitive means of analyzing hair digests; (ii) an analysis of artifact peptides arising from side reactions is also made using a new method for finding unexpected modifications; (iii) detailed analysis of the gel‐based method employed clearly shows the high degree of cross‐linking or protein association involved in hair digestion, with major GVPs eluting over a wide range of high molecular weights while others apparently arise from distinct non‐cross‐linked proteins; and (v) finally, we show that some of the specific GVP identifications depend on the sample preparation method.

Most DNA evidence is a mixture of two or more people. Cybergenetics TrueAllele® system uses Bayesian computing to separate genotypes from mixture data and compare genotypes to calculate likelihood ratio (LR) match statistics. This validation study examined the reliability of TrueAllele computing on laboratory‐generated DNA mixtures containing up to ten unknown contributors. Using log(LR) match information, the study measured sensitivity, specificity, and reproducibility. These reliability metrics were assessed under different conditions, including varying the number of assumed contributors, statistical sampling duration, and setting known genotypes. The main determiner of match information and variability was how much DNA a person contributed to a mixture. Observed contributor number based on data peaks gave better results than the number known from experimental design. The study found that TrueAllele is a reliable method for analyzing DNA mixtures containing up to ten unknown contributors.

Forensic examiners must determine whether both latent fingerprint development and DNA profiling can be performed on the same area of an evidence item and, if only one is possible, which examination offers the best chance for identification. Latent fingerprints can be enhanced by targeting different components of fingerprint residues with sequential chemical treatments. This study investigated the effects of single-reagent and sequential latent fingerprint development processes on downstream DNA analysis to determine the point at which latent fingerprint development should be stopped to allow for DNA recovery. Latent fingerprints deposited on copy paper by one donor were developed using three sequential processes: 1,8-diazafluoren-9-one (DFO) → ninhydrin → physical developer (PD); 1,2-indanedione-zinc (IND-Zn) → ninhydrin → PD; and IND-Zn → ninhydrin → Oil Red O (ORO) → PD. Samples were examined after the addition of each chemical treatment. DNA was collected with cotton swabs, extracted, quantified, and amplified. DNA yields, peak heights, number of alleles obtained, and percentage of DNA profiles eligible for CODIS upload were examined. DNA profiles were obtained with varying degrees of success, depending on the number and type of treatments used for latent fingerprint development. The treatments that were found to be the least harmful to downstream DNA analysis were IND-Zn and IND-Zn/laser, and the most detrimental treatments were DFO, DFO/laser, and PD. In general, as the number of treatments increase, the opportunities for DNA loss or damage also increase, and it is preferable to use fewer treatments when developing latent fingerprints prior to downstream DNA processing.

Image segmentation is a fundamental precursor to quantitative image analysis. At present, no standardised methodology exists for segmenting images of fluorescent proxies for trace evidence. Experiments evaluated (i) whether manual segmentation is reproducible within and between examiners (with three participants repeatedly tracing three images) (ii) whether manually defining a threshold level offers accurate and reproducible results (with 20 examiners segmenting 10 images), and (iii) whether a global thresholding algorithm might perform with similar accuracy, while offering improved reproducibility and efficiency (16 algorithms tested). Statistically significant differences were seen between examiners’ traced outputs. Manually thresholding produced good accuracy on average (within ±1% of the expected values), but poor reproducibility (with multiple outliers). Three algorithms (Yen, MaxEntropy, and RenyiEntropy) offered similar accuracy, with improved reproducibility and efficiency. Together, these findings suggest that appropriate algorithms could perform thresholding tasks as part of a robust workflow for reconstruction studies employing fluorescent proxies for trace evidence.

Forensic pathologists’ decisions are critical in police investigations and court proceedings as they determine whether an unnatural death of a young child was an accident or homicide. Does cognitive bias affect forensic pathologists’ decision‐making? To address this question, we examined all death certificates issued during a 10‐year period in the State of Nevada in the United States for children under the age of six. We also conducted an experiment with 133 forensic pathologists in which we tested whether knowledge of irrelevant non‐medical information that should have no bearing on forensic pathologists’ decisions influenced their manner of death determinations. The dataset of death certificates indicated that forensic pathologists were more likely to rule "homicide" rather than "accident" for deaths of Black children relative to White children. This may arise because the base‐rate expectation creates an a priori cognitive bias to rule that Black children died as a result of homicide, which then perpetuates itself. Corroborating this explanation, the experimental data with the 133 forensic pathologists exhibited biased decisions when given identical medical information but different irrelevant non‐medical information about the race of the child and who was the caregiver who brought them to the hospital. These findings together demonstrate how extraneous information can result in cognitive bias in forensic pathology decision‐making.

A blind quality control (QC) program was successfully developed and implemented in the Toxicology, Seized Drugs, Firearms, Latent Prints (Processing and Comparison), Forensic Biology, and Multimedia (Digital and Audio/Video) sections at the Houston Forensic Science Center (HFSC). The program was put into practice based on recommendations set forth in the 2009 National Academy of Sciences report and is conducted in addition to accreditation required annual proficiency tests. The blind QC program allows HFSC to test its entire quality management system and provides a real‐time assessment of the laboratory’s proficiency. To ensure the blind QC cases mimicked real casework, the workflow for each forensic discipline and their evidence submission processes were assessed prior to implementation. Samples are created and submitted by the HFSC Quality Division to whom the expected answer is known. Results from 2015 to 2018 show that of the 973 blind samples submitted, 901 were completed, and only 51 were discovered by analysts as being blind QC cases. Implementation data suggests that this type of program can be employed at other forensic laboratories.

Respiratory pathogens have been detected in forensic investigations using multiple techniques; however, no study has examined the use of automated, nested, multiplex polymerase chain reaction (ANM‐PCR), commonly used in living patients, in the forensic setting. This retrospective study assessed the utility of ANM‐PCR in detecting respiratory pathogens in the pediatric forensic setting. Respiratory samples from 35 cases were tested for up to 20 respiratory pathogens. 51.4% of these cases yielded a positive ANM‐PCR result, 20% of which were considered the cause of or contributory to death. The most commonly detected pathogens were rhinovirus/enterovirus and respiratory syncytial virus, and these were the only pathogens determined to play a significant role in cause of death. The sampled sites and postmortem intervals tested did not affect the likelihood of a positive or negative test. ANM‐PCR panels are effective, affordable, and rapid ancillary tools in evaluating cause of death in the forensic pediatric population.

Crime scene investigators and laboratory analysts use chemical tests to detect and differentiate body fluids. Testing often requires a sample of the stain, and the chemicals may cause degradation of the fluid or interfere with subsequent tests. Colorimetric chemical tests do not differentiate between different types of the same fluid, such as venous and menstrual blood, and there is no presumptive test available to simultaneously differentiate several body fluids. In this study, we recorded ATR FT-IR spectra of venous and menstrual blood, semen, saliva, and breastmilk. Neat and simulated casework body fluid samples were analyzed on cotton, nylon, wood, paper, and glass substrates. Differences in fluid composition, including proteins and small molecules, resulted in spectral differences. Venous and menstrual blood is differentiated by the peak at 1039 cm−1 attributed to phosphoric acid found in menstrual blood. Peak intensity is influenced by the porosity and weave of the substrate fabric.

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.

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.

This study aimed to develop and to assess an algorithm to facilitate lip print visualization, and to digitally analyze lip prints on different supports, by superimposition. It also aimed to classify lip prints according to sex. A batch image processing algorithm was developed, which facilitated the identification and extraction of information about lip grooves. However, it performed better for lip print images with a uniform background. Paper and glass slab allowed more correct identifications than glass and the both sides of compact disks. There was no significant difference between the type of support and the amount of matching structures located in the middle area of the lower lip. There was no evidence of association between types of lip grooves and sex. Lip groove patterns of type III and type I were the most common for both sexes. The development of systems for lip print analysis is necessary, mainly concerning digital methods.

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.