Premium
Use of a Real Time PCR Assay to Assess the Effect of Pulsed Light Inactivation on Bacterial Cell Membranes and Associated Cell Viability
Author(s) -
Garvey Mary,
Stocca Alessia,
Rowan Neil
Publication year - 2016
Publication title -
water environment research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.356
H-Index - 73
eISSN - 1554-7531
pISSN - 1061-4303
DOI - 10.2175/106143016x14504669767210
Subject(s) - membrane , real time polymerase chain reaction , ultraviolet light , chemistry , viability assay , lysis , viable but nonculturable , cell membrane , polymerase chain reaction , cell , biophysics , dna , dna damage , microbiology and biotechnology , bacteria , biology , biochemistry , gene , genetics , photochemistry
Research into more rapid and effective means of disinfecting water has become necessary due to the recognition that not all pathogenic species are being removed by chemical means. There is an extent of research highlighting the benefits of pulsed light for the disinfection of water. This study aims to determine the ability of a real time polymerase chain reaction assay to evaluate microbial inactivation of pulsed light treated cells. Findings show that pulsed light is a more rapid means of inactivating test species than standard UV lamp systems. A linear relationship between cell number and polymerase chain reaction amplification was obtained. A difference in threshold value (Ct) of approximately 4 ( p ≤ 0.05) was obtained for DNA amplification following the addition of the dye for pulsed ultrviolet (PUV)‐treated Bacillus cells. Membrane protein leakage proved an effective means of determining membrane damage for both Bacillus and E. coli test species following PUV treatment. This membrane damage was not evident for cells exposed to low pressure ultraviolet (LPUV). Findings describe suggest that PUV treatment induced a viable but nonculturable state in treated cells.