Evaluating Bioinformatic Pipeline Performance for Forensic Microbiome Analysis*,†,‡

Microbial communities have potential evidential utility for forensic applications. However, bioinformatic analysis of high-throughput sequencing data varies widely among laboratories. These differences can potentially affect microbial community composition and downstream analyses. To illustrate the importance of standardizing methodology, we compared analyses of postmortem microbiome samples using several bioinformatic pipelines, varying minimum library size or minimum number of sequences per sample, and sample size. Using the same input sequence data, we found that three open-source bioinformatic pipelines, MG-RAST, mothur, and QIIME2, had significant differences in relative abundance, alpha-diversity, and beta-diversity, despite the same input data. Increasing minimum library size and sample size increased the number of low-abundant and infrequent taxa detected. Our results show that bioinformatic pipeline and parameter choice affect results in important ways. Given the growing potential application of forensic microbiology to the criminal justice system, continued research on standardizing computational methodology will be important for downstream applications.     

Ethyl Glucuronide Concentration in Hair of Detainees: A Preliminary Study

Through the measurement of ethyl glucuronide in hair (hETG), it is possible to assess chronic alcohol abuse over time. In this paper, we present a study on hETG in Italian prison inmates. Analyses were performed by LC-MS according to a previously published method. Results were evaluated using the cut-offs established by the Society of Hair Testing. Positives samples (ETG > 30 pg/mg) accounted for 6% of all subjects, with concentrations ranging from 42 pg/mg up to 270 pg/mg, abstinent subjects (ETG < 7 pg/mg) accounted for 88%, and moderate alcohol consumption (7 < ETG < 30 pg/mg) for 6% of the subjects. No females displayed ETG values above 30 pg/mg. Among positive samples, only two subjects did not declare heavy alcohol consumption and were found strongly positive at 210 and 270 pg/mg. To the best of our knowledge, this represents the first study on ETG hair concentration on prison inmates.     

A Study of the Background Levels of Male DNA on Underpants Worn by Females

This study is the first to examine the background level of male DNA on underpants worn by females in the absence of sexual contact. Here, we examined 103 samples from the inside front of underpants from 85 female volunteers. Samples were examined for the presence of male DNA using NGM SElect and PowerPlex Y23 kits. Only five samples gave a “complete” Y-STR profile, even though 83.5% of our volunteers cohabited with a male. In all cases where a partner reference sample was available, the Y-STR profile matched the cohabiting partner. We have demonstrated that a Y-STR profile is not expected on the inside front of underpants worn by females after social contact alone. The results of this study are informative for evaluating the significance of a Y-STR profile on underpants in cases of alleged sexual assault.     

Development of a Novel Finger-Trigger Interface for Trigger Pull Measurement

Trigger pull is the force that needs to be exerted on the trigger to discharge a firearm. The measurement of trigger pull can assist in the evaluation of the safety, function, and manufacturing characteristics associated with a firearm during the forensic firearm examination process. Nonetheless, the accuracy and uncertainty of trigger pull measurements may be affected by the measuring device, test procedure, and environmental conditions. In this work, an innovative finger-trigger interface device was developed to facilitate accurate trigger pull measurements. The idea was to reduce the variation related to the position of the measurement device on the trigger in existing measuring methods and devices. Three force sensors based on different technologies were initially evaluated. While two of the three sensors failed to produce data, the miniature capacitive plate sensor exhibited high precision and a linear response over the range of typical trigger pulls. To examine the effects of the finger-trigger interface on trigger pull measurement, different sensor housing prototypes were designed in silico and 3D printed for the construction of three finger-trigger interface devices. The performance of each finger-trigger interface device was evaluated by measuring the trigger pulls of several selected firearms and comparing the data to a previously published study. Our preliminary results demonstrated the novel finger-trigger interface device offered a new way to measure trigger pull in situ with acceptable accuracy and precision.     

Improved Area of Origin Estimation for Bloodstain Pattern Analysis Using 3D Scanning

An important component of crime scene reconstruction is bloodstain pattern analysis (BPA). Where BPA concerns impact patterns, estimating the area of origin is critical information for scene reconstruction. Traditionally, this is achieved by measuring individual bloodstains and performing trigonometric calculations; however, 3D scanning has been proposed as a viable alternative for overcoming logistical and practical concerns with the manual method. Therefore, this project aimed to establish whether the FARO Focus 3D scanner and FARO Zone 3D software can improve the accuracy of area of origin estimates relative to the manual method. We created a series of eight bloodstain impact patterns and performed paired analysis using the two methods to estimate areas of origin for each pattern. Our data suggested that FARO-derived estimates were generally more accurate than using the manual method. FARO-estimated heights of origin areas were generally closer to the true distance. Both methods underestimated the distance from the wall for most patterns originating 150mm or greater from the wall, but overestimated distances for patterns originating closer to the wall. The degree to which distances were underestimated increased significantly the further the blood source was from the wall and was greater for FARO-derived estimates. The results of this research contribute to the validation of these instruments for operational implementation for BPA and should be considered alongside the practical benefits of 3D scanning relative to manual methods. Further, 3D scanning can provide reliable BPA reconstruction documentation for technical review and court presentation.     

Juvenile and Family Court Judges’ Knowledge and Attitudes About Sex Trafficking of Minors: Associations with Gender,Race, and U.S. Region

Juvenile and family court judges are a professional group that have a significant amount of decision-making power in cases of sex trafficking of minors. The purpose of this project is to examine the association of juvenile and family court judges’ gender, race, and U.S. region with their attitudes and knowledge about sex trafficking of minors. Drawing from a survey of 55 juvenile and family court judges in the U.S., this study used standardized scales to measure attitudes and knowledge about child sex trafficking. Results indicate some differences by gender and geography in a sample of experienced judges across the U.S. The consistency of these findings are discussed in the context of other research and implications for targeted training.     

Exploring Gender-Specific Trends in Underage Drinking Across Adolescent Age Groups and Measures of Drinking: Is Girls’ Drinking Catching Up with Boys’?

Underage drinking is among the most serious of public health problems facing adolescents in the United States. Recent concerns have centered on young women, reflected in media reports and arrest statistics on their increasing problematic alcohol use. This study rigorously examined whether girls’ alcohol use rose by applying time series methods to both arrest data, Uniform Crime Reports, and self-report data from Monitoring the Future, a nationally representative long-term survey gathered independently of crime control agents. All self-reported drinking behaviors across all age groups show declining or unchanged female rates and no significant change in the gender gap, while the official source displays a steady narrowing gender gap and some increase of female arrest rates for liquor law violations. Results indicate that social control measures applied to underage drinking have shifted to target young women’s drinking patterns, but their drinking has not become more widespread/problematic. Girls’ increased alcohol use and abuse is a socially constructed problem, rather than the result of normalization of drinking or more strain in girls’ lives. Future underage drinking policies and practices that apply legal intervention strategies to less chronic adolescent drinking behaviors will increase the visibility of girls’ drinking.