Dual Detection of the Chytrid Fungi Batrachochytrium spp. with an Enhanced Environmental DNA Approach
Abstract
:1. Introduction
2. Materials and Methods
2.1. Study Area and Collection of Field Samples
2.2. DNA Extraction and qPCR
2.3. gBlock In Vitro Testing
2.4. Storage Experiment for eDNA Samples
2.5. Cost Assessment
2.6. Statistical Analyses
3. Results
3.1. Field Sampling
3.2. gBlock In Vitro Testing
3.3. Storage Experiment for eDNA Samples
3.4. Cost Assessment
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Country | Locality | First Bsal Positive | N Filters | N Swabs | Bsal DNA Copies | Bd (DNA Copies) |
---|---|---|---|---|---|---|
Spain | Ampuero | * 2017 | 1/1 | 5/5 | 10.81 ‡/8.05–23 § | |
Spain | Teverga 1 | * 2017 | 1/1 | 1/19 | 96.55 ‡/15.05 § | |
Spain | Teverga 2 | * 2017 | 1/1 | 0 | 15.70 ‡ | |
Spain | Teverga 3 † | 2019 | 2/2 | 2/2 | 12.2–12.45 ‡/3.27–17 § | |
Spain | Ruente 1 | * 2017 | 1/2 | 0/20 | 12 ‡ | |
Spain | Ruente 2 | 2019 | 1/1 | 0/11 | 6.16 ‡ | 28.8 ‡ |
Spain | Suances | * 2017 | 2/2 | 0/25 | 22.35–96.05 ‡ | 31.2; 764.4 ‡ |
Spain | Ponga † | 2019 | 2/2 | 0/15 | 1.51–24.2 ‡ | |
Spain | Cieza | 2019 | 1/1 | 0/11 | 33.3 ‡ | |
Czech R. | Stare Mesto | N/A | 2/2 | 11/30 | N/A | 0.1–7.3 ‡/1.91–44.55 § |
Czech R. | Sokolov | N/A | 0/32 | N/A | N/A |
Storage Type | Primers from Blooi et al., 2013 | Primers from Spitzen-van der Sluijs et al., 2020 |
---|---|---|
Control, room temperature | 0.01% * | 0.1% * |
Longmire’s buffer | 0.04% | 0.2% * |
Silica gel | 0.19% | 1.1% |
96% EtOH | 0.34% | 1.1% |
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Lastra González, D.; Baláž, V.; Vojar, J.; Chajma, P. Dual Detection of the Chytrid Fungi Batrachochytrium spp. with an Enhanced Environmental DNA Approach. J. Fungi 2021, 7, 258. https://doi.org/10.3390/jof7040258
Lastra González D, Baláž V, Vojar J, Chajma P. Dual Detection of the Chytrid Fungi Batrachochytrium spp. with an Enhanced Environmental DNA Approach. Journal of Fungi. 2021; 7(4):258. https://doi.org/10.3390/jof7040258
Chicago/Turabian StyleLastra González, David, Vojtech Baláž, Jiří Vojar, and Petr Chajma. 2021. "Dual Detection of the Chytrid Fungi Batrachochytrium spp. with an Enhanced Environmental DNA Approach" Journal of Fungi 7, no. 4: 258. https://doi.org/10.3390/jof7040258