ATP Bioluminescence for Rapid and Selective Detection of Bacteria and Yeasts in Wine

Abstract

Microbial contamination may represent a loss of money for wine producers as several defects can arise due to a microorganism’s growth during storage. The aim of this study was to implement a bioluminescence assay protocol to rapidly and simultaneously detect bacteria and yeasts in wines. Different wines samples were deliberately contaminated with bacteria and yeasts at different concentrations and filtered through two serial filters with decreasing mesh to separate bacteria and yeasts. These were resuscitated over 24 h on selective liquid media and analyzed by bioluminescence assay. ATP measurements discriminated the presence of yeasts and bacteria in artificially contaminated wine samples down to 50 CFU/L of yeasts and 1000 CFU/L of bacteria. The developed protocol allowed to detect, rapidly (24 h) and simultaneously, bacteria and yeasts in different types of wines. This would be of great interest for industries, for which an early detection and discrimination of microbial contaminants would help in the decision-making process.

Oxygen Sensor-Based Respirometry and the Landscape of Microbial Testing Methods as Applicable to Food and Beverage Matrices

“…Additionally, food samples, such as meat or milk, can contain somatic cell ATP, which can cause interference and high background levels of BL. In order to detect microbial ATP, pre-treatment of samples is required [15,18]. Subsequently, to convert RLU signals into CFU/g, a standard curve must be established using known standards of bacteria [15].…”

“…ATP BL was also applied to the detection of yeast and bacteria in wine; however, special treatment of samples was required. Wine samples were filtered through 2 sets of membrane filters, which were then incubated in selective media for 24 h with added ATPase (hydrolyses free ATP) and analyzed via luminometer to eventually discern between varying levels of contamination with yeast and bacteria [18].…”

“…BL detection suffers from matrix effects and requires clear samples (diluted or processed). Therefore, lengthy steps of sample clarification and enrichment are often used [15,18]. DNA/RNA amplification assays have long and tedious sample preparation steps that include enrichment of target cells, cell lysis and RNA extraction and purification followed by RT-PCR.…”

Metabolomics of Pseudomonas aeruginosa Treated by Atmospheric-Pressure Cold Plasma

“…Samples were taken from the control group and the treatment group, and the reagent was injected into the reaction tube and had full contact with the swab. After the complete reaction, the swab was put into the ATP detector to detect the ATP content of bacteria [31,32].…”

Breaking the Mold: Towards Rapid and Cost-Effective Microbial Contamination Detection in Paints and Cosmetics Using ATP-Bioluminescence

“…Further limitations include possible preceding enrichment steps and the choice of media and/or incubation conditions that orientate the microbial groups to be cultivated. Most importantly, many microorganisms are not (yet) cultivable or exist in a viable but non-cultivable (VBNC) state, allowing microbial contaminations to arise quickly and without any warning [3]. To comply with those requirements, culture-independent methods have been recognized as a promising alternative to be used in conjunction with, or even supplanting, culture-dependent methods [4].…”