A simple bioluminescence procedure for early warning detection of coliform bacteria in drinking water

Abstract

Traditional cultivation-dependent tests for coliform bacteria in food and drinking water take 18-24 h to complete. Bioluminescence-based enzyme assays can potentially reduce analysis time for indicator bacteria such as coliforms. In the present study, we developed a simple presence/absence (P/A) bioluminescence procedure for rapid detection of coliform bacteria in groundwater-based drinking water. The bioluminescence procedure targeting b-D-galactosidase activity in coliform bacteria was based on hydrolysis of 6-O-b-galactopyranosyl-luciferin. Bacteria immobilized on membrane filters were enriched for 6-8 h in selective media containing isopropyl-b-D-thiogalactopyranoside (IPTG) to induce b-D-galactosidase activity in coliform bacteria. The equivalent of approximately 300 E. coli cells was required for bioluminescence detection of b-D-galactosidase activity. In comparison, PCR based detection of E. coli in drinking water required approximately 30 target cells. Analysis of contaminated drinking water samples showed comparable results for coliform bacteria using traditional multiple-tube fermentation, Colilert-18, and the bioluminescence procedure. Aeromonas hydrophila or indigenous groundwater bacteria did not interfere with the procedure. The bioluminescence procedure can be combined with commercial substrates such as Fluorocult or Colilert-18, and will allow the detection of one coliform in 100 ml drinking water within one working day. The results suggest the bioluminescence assays targeting b-D-galactosidase activity may be used for or for early warning screening of drinking water and/or rapid identification of contaminated drinking water wells.

An Intact Cell Bioluminescence-Based Assay for the Simple and Rapid Diagnosis of Urinary Tract Infection

“…Bioluminescent bacteria are prokaryotes that can produce their own light solely from their unique metabolic activity. Marine luminous bacteria have been studied and extensively utilized in the biotechnological detection of toxic chemicals, pollutants, pesticides, dissolved oxygen, freshness, quorum sensing, ATP, and other physicochemical conditions [17][18][19][20][21][22][23]. Bioluminescence is living light; the partial or complete inhibition depends on the cells' viability, which is affected by the cells' immediate physicochemical condition and/or direct exposure to toxicants.…”

“…Bioluminescence is living light; the partial or complete inhibition depends on the cells' viability, which is affected by the cells' immediate physicochemical condition and/or direct exposure to toxicants. Thus, bioluminescence lends itself well as a biological sensing tool for the direct observation and measurement of cell viability and well-being [18,19,24]. In addition, it contributes to important applications in other scientific disciplines, particularly in the field of diagnostic medicine.…”

Amperometric detection of Enterobacteriaceae in river water by measuring β-galactosidase activity at interdigitated microelectrode arrays

“…In the case of coliforms, this enzyme is ␤-galactosidase (␤-gal). The whole procedure is rather long and the analysis time ranges from 18 to 24 h. Efforts to detect these microorganisms faster have led to the development of new methods based on fluorescence [2], bioluminescence [3], PCR [4] and biosensing [5,6].…”

Relations fecal coliforms/ fecal Streptococci as indicators of the origin of fecal pollution in urban and rural water bodies of Temuco, Chile

“…The presence of these bacteria is considered as an index of fecal contamination and therefore contamination with pathogenic organisms (Sankararamakrishnan and Guo, 2005;Bastholm et al, 2008).…”