DNA Barcoding for Molecular Prospecting of Platyhelminthes

DNA barcoding (Folmer et al. 1994), is an efficient method to distinguish species by short specific DNA sequences from a common region of their genome, such as the cytochrome c oxidase subunit 1 (CO1) gene. The early primers of Folmer et al. (1994) were not as universal as desired (Elias et al. 2007). Platyhelminthes (flatworms), being the fourth most speciose animal phylum, is a taxonomic group where universal barcoding primers are not very effective (Vanhove, et al. 2013). Recently, Van Steenkiste et al. (2014) and Elbrecht and Leese (2017) developed primers for parasitic and free-living flatworms that show promise as molecular barcodes. We tested the primers developed by Van Steenkiste et al. (2014), Dice 1F, Dice 11R, and Dice 14R, on a diverse collection of trematodes, a group with a significant number of undescribed taxa. A total of 120 amplifications were performed on 69 trematode samples from 27 genera. Dice 1F/11R and Dice 1F/14R primer sets were tested alongside the JB3/JB5 primer set (Bowles et al. 1992; Derycke et al. 2005). Overall amplification efficacy was notably larger for the Dice 1F/11R primer set. However, the JB3/JB5 primer pair led to a higher percentage of successful sequences, as compared to either of the Dice primers.https://digitalcommons.snc.edu/collaborative_presentations/1061/thumbnail.jp

Antimicrobial Effects of Compounds Used in Cleaning Wipes

The antimicrobial effectiveness and level of biofilm inhibition of three compounds from Rockline Industries were tested, along with five compounds made by the St. Norbert College Organic Chemistry Department. The Rockline compounds, ColaLipid (CL), Sodium Benzoate (NaB) and Benzoic Acid (BA), were solubilized in 10% DMSO and tested at concentrations of 0.15%, 0.10%, and 0.01%. To ensure that any growth effects were due to the compounds in question, DMSO was also tested for antimicrobial activity. The compounds were tested against safe relative of eight bacteria of clinical importance (Acinetobacter baylyi, Bacillus subtilis, Escherichia coli, Enterococcus raffinosus, Enterobacter aerogenes, Pseudomonas fluorescens, Pseudomonas putida, Staphylococcus epidermidis) and one species of fungus (Candida albicans). The antimicrobial properties of these compounds were measured by reading absorbance at 600 nm of overnight cultures. Biofilm formation inhibition was measured using a crystal violet biofilm assay. ColaLipid and Benzoic Acid were found to inhibit growth of the bacterial and fungal strains, while Sodium Benzoate did not. Of the compounds synthesized by in St. Norbert organic chemists, two showed consistent inhibition of growth and three did not. ColaLipid, Benzoic Acid, and one organic compound showed consistent inhibition of biofilm formation.https://digitalcommons.snc.edu/collaborative_presentations/1081/thumbnail.jp