Selection and Evaluation of Staphylococcus xylosus as a Biocontrol Agent against Toxigenic Moulds in a Dry-Cured Ham Model System
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacteria and Fungi
2.2. Staphylococcal Enterotoxin Analysis
2.3. 16S rDNA Sequence Analysis
2.4. Inocula Preparation
2.5. Effect of GCC+ on Growth of Ochratoxigenic Moulds
2.6. Effect of S. xylosus on Growth and Mycotoxins Production by Mycotoxigenic Moulds
2.6.1. Growth Assessment of Toxigenic Moulds
2.6.2. Extraction and Quantification of Mycotoxins
OTA Extraction
Aflatoxins Extraction
CPA Extraction
Recovery Experiments and Linearity Range
Mycotoxin Quantification by Orbitrap Q Exactive Plus Analysis
Mycotoxin Determination Sensitivity
2.7. Statistical Analysis
3. Results
3.1. Enterotoxigenic Potential of Gram-Positive, Catalase-Positive Cocci
3.2. Selection and Identification of Gram-Positive, Catalase-Positive Cocci Isolates with Antifungal Activity
3.3. Antifungal Activity of S. xylosus in a Dry-Cured Ham Model System
3.3.1. Effect on Mould Growth
3.3.2. Effect on Mycotoxin Production
4. Discussion
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Isolate | Identification | Accession Number | % Identity |
---|---|---|---|
Ku13 | K. uropygioeca | NR_157676.1 | 95.88 |
Ku15 | K. uropygioeca | NR_157676.1 | 98.72 |
Ku40 | K. uropygioeca | NR_157676.1 | 100 |
Se28 | S. equorum | MK015791.1 | 99.86 |
Se29 | S. equorum | MN229550.1 | 99.79 |
Se31 | S. equorum | MN229551.1 | 99.91 |
Se32 | S. equorum | MN758799.1 | 98.28 |
Se39 | S. equorum | KY940339.1 | 99.85 |
Se41 | S. equorum | MK796070.1 | 99.92 |
Se47 | S. equorum | MN758799.1 | 99.64 |
Se50 | S. equorum | KP224447.1 | 99.56 |
Se53 | S. equorum | KY940339.1 | 99.86 |
Ss21 | S. saprophyticus | KU922358.1 | 99.02 |
Ss42 | S. saprophyticus | MF457583.1 | 99.4 |
Ss46 | S. saprophyticus | MF457583.1 | 99.7 |
Ss7 | S. saprophyticus | MN229561.1 | 98.52 |
Ss9 | S. saprophyticus | KT717631.1 | 99 |
Sv33 | S. vitulinus | KM378591.1 | 99.69 |
Sw16 | S. warneri | MK414943.1 | 98.8 |
Sx8 | S. xylosus | HM854231.1 | 99 |
S17 | S. pasteuri/S. warneri | MH930440.1/MN181250.1 | 99.79 |
S19 | S. pasteuri/S. warneri | MH930440.1/MN421516.1 | 99.79 |
15 °C | 20 °C | 25 °C | ||||
---|---|---|---|---|---|---|
Mould | Control | Sx8 | Control | Sx8 | Control | Sx8 |
Pn15 | 3.93 ± 1.36 | <LOQ *,a | 18.76 ± 3.22 | <LOQ * | 66.59 ± 19.52 | <LOQ * |
Pn92 | 2.75 ± 0.49 | <LOD *,b | 26.61 ± 4.82 | <LOD * | 89.53 ± 14.90 | <LOD * |
Pn856 | 31.54 ± 0.07 | 4.23 ± 0.35 * | 260.64 ± 71.93 | 64.26 ± 11.72 * | 389.73 ± 57.65 | 860.13 ± 357.03 * |
15 °C | 20 °C | 25 °C | ||||
---|---|---|---|---|---|---|
Mould | Control | Sx8 | Control | Sx8 | Control | Sx8 |
Af65 | 2.09 ± 0.25 | 0.18 ± 0.03 * | 41.06 ± 6.23 | 0.28 ± 0.09 * | 28.49 ± 3.07 | 2.08 ± 2.21 * |
Ap2681 | 0.08 ± 0.02 | 0.01 ± 0.01 * | 0.06 ± 0.05 | 0.01 ± 0.01 | 0.10 ± 0.02 | <LOD *,a |
15 °C | 20 °C | 25 °C | ||||
---|---|---|---|---|---|---|
Mould | Control | Sx8 | Control | Sx8 | Control | Sx8 |
Ap2681 | 0.73 ± 0.34 | <LOD *,a | 4.85 ± 8.24 | <LOD * | 0.91 ± 0.30 | <LOD * |
15 °C | 20 °C | 25 °C | ||||
---|---|---|---|---|---|---|
Mould | Control | SX8 | Control | SX8 | Control | SX8 |
Pg2919 | 1270.4 ± 431.9 | 888.5 ± 258.3 | 2427.8 ± 724.8 | 1793.9 ± 34.7 * | 4492.9 ± 227.0 | 2694.1 ± 881.9 * |
Pg14319 | 2804.2 ± 802.8 | 1160.0 ± 165.0 * | 4120.4 ± 944.7 | 2932.9 ± 651.9 | 6298.7 ± 2.9 | 5358.9 ± 382.4 * |
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Cebrián, E.; Núñez, F.; Gálvez, F.J.; Delgado, J.; Bermúdez, E.; Rodríguez, M. Selection and Evaluation of Staphylococcus xylosus as a Biocontrol Agent against Toxigenic Moulds in a Dry-Cured Ham Model System. Microorganisms 2020, 8, 793. https://doi.org/10.3390/microorganisms8060793
Cebrián E, Núñez F, Gálvez FJ, Delgado J, Bermúdez E, Rodríguez M. Selection and Evaluation of Staphylococcus xylosus as a Biocontrol Agent against Toxigenic Moulds in a Dry-Cured Ham Model System. Microorganisms. 2020; 8(6):793. https://doi.org/10.3390/microorganisms8060793
Chicago/Turabian StyleCebrián, Eva, Félix Núñez, Fernando J. Gálvez, Josué Delgado, Elena Bermúdez, and Mar Rodríguez. 2020. "Selection and Evaluation of Staphylococcus xylosus as a Biocontrol Agent against Toxigenic Moulds in a Dry-Cured Ham Model System" Microorganisms 8, no. 6: 793. https://doi.org/10.3390/microorganisms8060793