ATP Bioluminescence for Rapid and Selective Detection of Bacteria and Yeasts in Wine
Abstract
:1. Introduction
2. Materials and Methods
2.1. Wine Samples
2.2. Bacterial Strains and Deliberate Wine Contamination
2.3. Bioluminescence Assay
2.4. Plating
3. Results
4. Discussion
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Wine Samples | Sparkling | Alcohol (%) | Sugar (g/L) |
---|---|---|---|
R1 | No | 13.00 | 3.7 |
R2 | No | 13.00 | 1 |
R3 | No | 13.00 | 3.1 |
R4 | No | 6.00 | 125 |
R5 | Yes | 6.00 | 110 |
R6 | Yes | 12.00 | 15 |
W1 | No | 12.00 | 3.5 |
W2 | No | 13.00 | 23.9 |
W3 | Yes | 5.00 | 125 |
W4 | Yes | 11.00 | 13 |
W5 | Yes | 6.00 | 110 |
W6 | Yes | 12.00 | 5 |
W7 | Yes | 8.00 | 80 |
W8 | Yes | 12.00 | 10 |
R7 | Yes | 12.00 | 10 |
V1 | No | 15.00 | 152 |
V2 | No | 15.00 | 168 |
Pass/Fail Limits | RLU Value |
---|---|
Sterile | ≤2 times blank |
Re-test | 2 times blank < RLU value < 3 times blank |
Non-sterile | ≥3 times blank |
Inoculum (CFU/mL) | Yeast (104) + Bacteria (107) | Yeast (104) + Bacteria (103) | ||||||
---|---|---|---|---|---|---|---|---|
Filter | 1.2 µm | 0.45 µm | 1.2 µm | 0.45 µm | ||||
Analysis | RLU YPDch | SD | RLU MRScy | SD | RLU YPDch | SD | RLU MRScy | SD |
R1 | 7.04 × 106 | 2.24 × 104 | 1.61 × 106 | 6.03 × 104 | 9.23 × 105 | 8.84 × 104 | 1.01 × 103 | 2.91 × 10 |
R2 | 3.86 × 106 | 3.32 × 104 | 1.14 × 106 | 2.24 × 104 | 7.66 × 106 | 2.14 × 104 | 1.01 × 105 | 4.28 × 103 |
R3 | 4.28 × 106 | 2.96 × 104 | 1.43 × 106 | 6.24 × 104 | 9.40 × 106 | 4.05 × 104 | 1.11 × 103 | 2.36 × 102 |
R4 | 3.02 × 106 | 2.13 × 104 | 9.23 × 105 | 1.90 × 104 | 7.38 × 106 | 4.11 × 104 | 1.14 × 103 | 8.55 × 10 |
R5 | 1.29 × 106 | 1.89 × 103 | 4.11 × 105 | 4.83 × 103 | 3.75 × 106 | 1.87 × 104 | 1.37 × 106 | 9.89 × 103 |
R6 | 1.45 × 106 | 4.88 × 104 | 9.58 × 105 | 5.23 × 104 | 4.84 × 106 | 3.76 × 104 | 5.70 × 105 | 1.57 × 104 |
W1 | 5.25 × 106 | 5.91 × 104 | 1.32 × 106 | 7.58 × 104 | 6.80 × 106 | 1.97 × 104 | 1.57 × 104 | 1.45 × 102 |
W2 | 3.96 × 106 | 1.20 × 104 | 4.91 × 105 | 7.99 × 103 | 7.35 × 106 | 5.69 × 104 | 1.02 × 103 | 8.59 × 10 |
W3 | 4.34 × 106 | 4.05 × 104 | 1.47 × 106 | 4.48 × 104 | 8.07 × 106 | 6.91 × 104 | 1.24 × 104 | 1.41 × 102 |
W4 | 9.69 × 106 | 3.19 × 104 | 1.33 × 106 | 6.91 × 104 | 3.85 × 106 | 3.01 × 104 | 1.31 × 106 | 2.82 × 104 |
W5 | 3.91 × 106 | 1.63 × 104 | 2.19 × 104 | 5.05 × 103 | 7.75 × 106 | 9.16 × 104 | 1.80 × 105 | 2.48 × 103 |
W6 | 1.74 × 106 | 1.44 × 104 | 4.93 × 105 | 1.04 × 104 | 8.04 × 106 | 5.14 × 104 | 5.39 × 105 | 2.28 × 103 |
Blank | 4.24 × 103 | 6.55 × 102 | 3.65 × 103 | 3.16 × 102 | 4.04 × 103 | 1.10 × 103 | 2.03 × 103 | 7.12 × 102 |
Inoculum (CFU/mL) | Filter | Analysis | R1 | R2 | R3 | R4 | R5 | R6 | W1 | W2 | W3 | W4 | W5 | W6 | TtD |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Yeast (104) + Bacteria (107) | 1.2 µm | RLU YPDch 1 | + | + | + | + | + | + | + | + | + | + | + | + | 24 h |
plating RLU-YPDch on YPDch agar 2 | + | + | + | + | + | + | + | + | + | + | + | + | 48 h | ||
Eluate on YPDch agar | − | − | − | − | − | − | − | − | − | − | − | − | |||
Eluate on MRScy agar | + | + | + | + | + | + | + | + | + | + | + | + | |||
0.45 µm | RLU MRScy 1 | + | + | + | + | + | + | + | + | + | + | + | + | 24 h | |
Plating RLU-MRScy on MRScy agar 2 | + | + | + | + | + | + | + | + | + | + | + | + | 48 h | ||
Yeast (104) + Bacteria (103) | 1.2 µm | RLU YPDch | + | + | + | + | + | + | + | + | + | + | + | + | 24 h |
plating RLU-YPDch on YPDch agar | + | + | + | + | + | + | + | + | + | + | + | + | 48 h | ||
Eluate on YPDch agar | − | − | − | − | − | − | − | − | − | − | − | − | |||
Eluate on MRScy agar | − | − | − | − | + | − | − | − | + | − | + | − | |||
0.45 µm | RLU MRScy | − | + | − | − | + | + | + | − | + | + | + | + | 24 h | |
Plating RLU-MRScy on MRScy agar | − | − | − | − | + | + | + | − | + | + | + | + | 48 h |
Inoculum (CFU/L) | Yeast (50) + LAB (1000) | |||||||
---|---|---|---|---|---|---|---|---|
Filter | 1.2 µm | 0.45 µm | ||||||
Res.Medium | YPDch | SD | RESch | SD | MRScy | SD | REScy | SD |
W7 | 1.93 × 106 | 2.08 × 102 | 4.35 × 105 | 2.74 × 104 | 2.89 × 103 | 4.08 × 102 | 1.22 × 104 | 4.83 × 103 |
W8 | 1.93 × 104 | 1.37 × 103 | 5.02 × 106 | 5.19 × 104 | 8.72 × 104 | 1.88 × 103 | 5.35 × 104 | 1.04 × 103 |
R7 | 1.92 × 105 | 3.29 × 104 | 6.81 × 106 | 4.09 × 103 | 1.73 × 106 | 1.75 × 104 | 7.20 × 104 | 2.13 × 103 |
V1 | 1.77 × 106 | 9.21 × 105 | 6.81 × 106 | 1.24 × 106 | 2.15 × 102 | 1.02 × 105 | 7.28 × 102 | 6.02 × 102 |
V2 | 3.47 × 102 | 2.86 × 102 | 5.59 × 106 | 2.75 × 105 | 7.07 × 102 | 7.62 × 102 | 5.78 × 103 | 6.84 × 102 |
Blank | 2.37 × 102 | 3.46 × 10 | 1.35 × 102 | 2.43 × 10 | 1.73 × 102 | 2.26 × 10 | 1.35 × 102 | 1.20 × 10 |
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Monica, S.; Bancalari, E.; Castellone, V.; Rijkx, J.; Wirth, S.; Jahns, A.; Bottari, B. ATP Bioluminescence for Rapid and Selective Detection of Bacteria and Yeasts in Wine. Appl. Sci. 2021, 11, 4953. https://doi.org/10.3390/app11114953
Monica S, Bancalari E, Castellone V, Rijkx J, Wirth S, Jahns A, Bottari B. ATP Bioluminescence for Rapid and Selective Detection of Bacteria and Yeasts in Wine. Applied Sciences. 2021; 11(11):4953. https://doi.org/10.3390/app11114953
Chicago/Turabian StyleMonica, Saverio, Elena Bancalari, Vincenzo Castellone, Jos Rijkx, Sarah Wirth, Anika Jahns, and Benedetta Bottari. 2021. "ATP Bioluminescence for Rapid and Selective Detection of Bacteria and Yeasts in Wine" Applied Sciences 11, no. 11: 4953. https://doi.org/10.3390/app11114953
APA StyleMonica, S., Bancalari, E., Castellone, V., Rijkx, J., Wirth, S., Jahns, A., & Bottari, B. (2021). ATP Bioluminescence for Rapid and Selective Detection of Bacteria and Yeasts in Wine. Applied Sciences, 11(11), 4953. https://doi.org/10.3390/app11114953