Date of Award
8-2024
Degree Type
Masters Thesis
Degree Name
Master of Science (MS)
School
Ocean Science and Engineering
Committee Chair
Kristina Mojica
Committee Chair School
Ocean Science and Engineering
Committee Member 2
Alan Shiller
Committee Member 2 School
Ocean Science and Engineering
Committee Member 3
Stephan Howden
Committee Member 3 School
Ocean Science and Engineering
Abstract
A multi-color flow cytometry method coupled with fluorescent staining was developed to detect and quantify E. faecalis concentrations in coastal waters as an alternative to the traditional membrane filtration method. The developed method was used to characterize E. faecalis during batch growth and yielded consistent results across the various physiological states of the cells. Additionally, the application of propidium iodide (i.e., a viability stain), afforded the method the ability to differentiate between cells with intact and compromised cell membranes, which raises the potential of quantifying cells that are in viable-but-non-culturable (VBNC) state. Initial field test using natural seawater samples spiked with E. faecalis, demonstrated high accuracy (98-120 % recovery rate) and precision (6-10% CV) of the method to detect E. faecalis. The short analysis time (~ 2 hours) and absence of labor-intensive steps (vacuum filtration, agar plating, and a series of confirmatory steps), increase the potential for the method to increase the throughput of routine monitoring of water quality. A shorter response time to potential outbreaks of E. faecalis and the ability to quantify VBNC cells would further reduce risks to public health and safety.
Copyright
2024, Riza Magbitang
Recommended Citation
Magbitang, Riza, "Development of a Flow Cytometry Method for the Identification and Enumeration of Enterococcus faecalis" (2024). Master's Theses. 1048.
https://aquila.usm.edu/masters_theses/1048
Included in
Analytical Chemistry Commons, Environmental Monitoring Commons, Marine Biology Commons, Water Resource Management Commons