Some microemulsions and nanoemulsions may have antimicrobial properties and be effective anti-biofilm agents. We examined the abilities of two fine emulsions, designated BCTP and TEOP, to inactivate suspensions of vegetative cells of Salmonella spp. Escherichia coli 0157:H7 (VT-), Pseudomonas aeruginosa, Staphylococcus aureus and Listeria monocytogenes. BCTP is an O/W nanoemulsion of soybean oil and tri-n-butyl phosphate emulsified with Triton X-100, while TEOP is an O/W microemulsion of ethyl oleate with Tween 80 as emulsifier and n-pentanol as a co-emulsifier. BCTP was effective in reducing the cell numbers of L. monocytogenes, while TEOP was effective against all five organisms investigated. The abilities of these emulsions to reduce preformed biofilms of the five bacteria were also investigated. With the exception of the biofilm formed by L. monocytogenes, which surprisingly was not significantly affected by BCTP, all biofilms were inhibited by both BCTP and TEOP.
Molecular studies, especially in relation to the activity of secondary metabolite gene clusters, require the ability to extract good quality RNA from fungal biomass. This is often hindered by the cell wall structure and endogenous RNase activity in filamentous fungi. There is thus a need for rapid methods for the extraction of good quality RNA for use in microarrays and for quantitative PCR assays. The objective of this study was to examine the use of different systems for the high throughput method to extract intact RNA from filamentous fungi. Two bead beating systems with different motion patterns and speed capacities were tested in the development of the extraction protocol. They were evaluated based on the total RNA yield and overall RNA quality. The high speed bead beating with glass beads associated with an automated purification method gave more than three times higher total RNA yields with less than a quarter of the amount of mycelium required. Furthermore the integrity and overall quality was conserved, with RNA Quality Indicator (RQI) numbers consistently >7.5. This method also reduced cross contamination risks and kept RNA handling to a minimum while still being capable of multiple sample processing, reducing the time required to obtain RNA from filamentous fungi.
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