Defense Date


Document Type

Directed Research Project

First Advisor

Dr. Sarah Seashols-Williams

Second Advisor

Dr. Baneshwar Singh

Third Advisor

Dr. Tomasz Arodz


The accurate identification of body fluids is vital for crime scene investigation and forensic analysis. While many body fluids can be presumptively identified, there is a lack of confirmatory testing for verification. Therefore, the purpose of this study was to determine if body fluids (menstrual blood, vaginal fluid, and semen) could be differentiated based on their eukaryotic microbial communities. The target area of interest was the V9 hyper-variable region of 18S ribosomal DNA (18S rDNA). The samples were extracted, barcoded using a duel-index strategy, and PCR amplified before undergoing high throughput sequencing using the Illumina Miseq FGx instrument. The samples were then analyzed using an open source platform called Mothur (v1.39.5) alongside the R (v3.6.3) program. Using an Analysis of Molecular Variance (AMOVA) test, the results showed that vaginal fluid and semen were significantly different from one another, but menstrual blood could not be differentiated from vaginal fluid or semen. From the relative abundance data, it was determined that the most abundant eukaryotic families in vaginal fluid were Debaryomycetaceae and Saccharomycetaceae while in semen it was Malasseziaceae. Menstrual blood appeared to show combinations of Debaryomycetaceae, Saccharomycetaceae, and Malasseziaceae. This data helped show that vaginal fluid and semen could potentially be used for the differentiation of body fluids in crime scenes. In the future, this study may be combined with the previous 16S rDNA study to develop a confirmatory test for the accurate identification of human body fluids.


© The Author(s)

Is Part Of

VCU Master of Science in Forensic Science Directed Research Projects

Date of Submission


Included in

Biology Commons


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