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.
Highlights
Microbial contamination may represent a consistent loss of money for wine producers, due to both yeasts and bacteria contaminating wines after primary fermentation
In the present study we developed a protocol, which coupled a two-filtration step with the bioluminescence assay, allowing for the simultaneous detection of yeasts and bacteria in wines
One filtration step through a 0.45 μm pore size filter would retain all microbial cells [18], hampering the possibility to discriminate between adenosine triphosphate (ATP) from yeasts or bacteria
Summary
Microbial contamination may represent a consistent loss of money for wine producers, due to both yeasts and bacteria contaminating wines after primary fermentation. Recognizing contaminations in time is fundamental to avoid economical losses but can be very tricky since a specific problem in wine can be provoked by different microorganisms such as lactic acid bacteria (LAB) or yeasts [2]. For these reasons it is important to develop a system that is faster than plating and able to discriminate between different sources of microbial contamination. Several molecular-based testing methods have been developed to detect spoilage microorganisms in wine, such as bio- and nanosensors, fluorescence cells sorting, and real-time polymerase chain reaction (RT-PCR). The choice of a method able to detect and distinguish the contamination from yeasts and bacteria would be of primary importance [13]
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