Acute Toxicity Assays with Adult Coral Fragments: A Method for Standardization
Abstract
:1. Introduction
2. Materials and Methods
2.1. Preparation of M. digitata Fragments
2.2. Endpoint Assessments
2.3. Preparation of Stock Solutions and Test Concentrations
2.4. Experimental Setup and Procedure
2.4.1. Water Sampling
2.4.2. Validation Criteria
2.5. Chemical Control Analyses
2.5.1. Analyses of Organic Test Substances: BP-3 and DCMU
2.5.2. Analyses of Inorganic Test Substance: CuCl2
2.6. Data Analysis
3. Results
3.1. Recovery of the Test Substances
3.2. Cu2+ Toxicity
3.3. BP-3 Toxicity
3.4. DCMU Toxicity
3.5. Effects of DMF as Carrier Solvent
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Validation Criteria | Target |
---|---|
Temperature | 24–28 °C, but constant within ±1.5 °C of cultivation temperature |
Salinity | 33–36 PSU |
pH | 7.7–8.5 |
O2 saturation | ≥75% * |
Alkalinity | 5–9 °dKH |
Bleaching intensity of control | ≤30% |
Fv/Fm of control | Fv/Fm ≥ 0.450 |
Tissue loss of control | ≤20% (1 fragment) |
Mortality of control | 0% |
Chem. | Nominal Conc. | Mean Measured Conc. of t0 to t96 | Recovery of Nominal | Mean Measured Conc. 1st Phase | Mean Measured Conc. 2nd Phase | Recovery t0–t48 | Recovery t48–t96 | Mean Recovery | ||
---|---|---|---|---|---|---|---|---|---|---|
t0 | t48 | t48 | t96 | Semi -static test method | ||||||
fresh | aged | fresh | aged | |||||||
mg L−1 | mg L−1 | % | mg L−1 | % | % | % | ||||
Control | 0 | <LoD | 100.0 | <LoD | <LoD | <LoD | <LoD | 100.0 | 100.0 | 100.0 |
BP-3 | 7.000 | 3.08700 | 44.1 | 3.37900 | 2.64100 | 3.91800 | 2.97300 | 78.2 | 75.9 | 77.1 |
8.000 | 4.29000 | 53.6 | 4.65600 | 3.70200 | 5.32800 | 4.17500 | 79.5 | 78.4 | 79.0 | |
9.000 | 5.43400 | 60.4 | 6.13600 | 4.62100 | 6.56200 | 5.33300 | 75.3 | 81.3 | 78.3 | |
10.000 | 6.38600 | 63.9 | 7.12000 | 5.37200 | 7.81600 | 6.31800 | 75.4 | 80.8 | 78.1 | |
11.000 | 7.33900 | 66.7 | 8.10800 | 5.92800 | 9.45100 | 7.31000 | 73.1 | 77.3 | 75.2 | |
12.000 | 9.07800 | 75.6 | 10.04900 | 7.39500 | 10.54600 | 9.54300 | 73.6 | 90.5 | 82.0 | |
Control | 0 | 0 | 100.0 | 0 | 0 | 0 | 0 | 100.0 | 100.0 | 100.0 |
0.00013 | 0.00018 | 139.1 | 0.00016 | 0.00018 | 0.00018 | 0.00017 | 111.5 | 93.0 | 102.2 | |
0.00064 | 0.00088 | 137.0 | 0.00081 | 0.00089 | 0.00085 | 0.00091 | 110.2 | 106.6 | 108.4 | |
0.00320 | 0.00441 | 137.7 | 0.00410 | 0.00442 | 0.00446 | 0.00452 | 107.6 | 101.3 | 104.4 | |
0.01600 | 0.01281 | 80.1 | 0.01150 | 0.00995 | 0.01607 | 0.01470 | 86.5 | 91.5 | 89.0 | |
DCMU | 0.08000 | 0.04937 | 61.7 | 0.04125 | 0.03875 | 0.05913 | 0.05918 | 93.9 | 100.1 | 97.0 |
0.40000 | 0.28860 | 72.2 | 0.22682 | 0.21678 | 0.36728 | 0.35198 | 95.6 | 95.8 | 95.7 | |
2.00000 | 1.91247 | 95.6 | 1.53252 | 1.53191 | 2.28243 | 2.29802 | 100.0 | 100.7 | 100.3 | |
10.00000 | 9.39022 | 93.9 | 7.06999 | 7.16035 | 11.55859 | 11.69601 | 101.3 | 101.2 | 101.2 | |
50.00000 | 40.93958 | 81.9 | 37.01764 | 34.42754 | 50.58926 | 44.58775 | 93.0 | 88.1 | 90.6 | |
Control | 0 | 0.00125 | 100.1 | 0 | 0 | 0 | 0.005 | 100.0 | 100.0 | 100.0 |
Cu2+ | 0.005 | 0.00200 | 40.0 | n.d. | 0 | 0.003 | 0.003 | n.d. | 100.0 | 100.0 |
0.009 | 0.00433 | 48.1 | n.d. | 0.003 | 0.005 | 0.005 | n.d. | 100.0 | 100.0 | |
0.019 | 0.00675 | 35.5 | 0.006 | 0.006 | 0.007 | 0.008 | 100.0 | 114.3 | 107.1 | |
0.038 | 0.01425 | 37.5 | 0.009 | 0.016 | 0.018 | 0.014 | 177.8 | 77.8 | 127.8 | |
0.075 | 0.01700 | 22.7 | 0.015 | n.d. | n.d. | 0.019 | n.d. | n.d. | n.d. | |
0.151 | 0.03150 | 20.9 | 0.023 | 0.023 | 0.042 | 0.038 | 100.0 | 90.5 | 95.2 | |
0.303 | n.d. | n.d. | n.d. | n.d. | n.d. | n.d. | n.d. | n.d. | n.d. |
Endpoint | Cu2+ | BP-3 | DCMU | DMF | |
---|---|---|---|---|---|
Mortality | LC50 | 0.161 (0.055–0.267) | 5.716 (5.256–6.176) | >9.390 | >1416 |
Tissue Loss | EC50 | 0.107 (0.076–0.138) | 5.112 (4.604–5.619) | >9.390 | >1416 |
Bleaching | EC50 | 0.008 (0.005–0.011) | >9.078 | 0.0275 (0.01750–0.03751) | >1416 |
Fv/Fm Loss | EC50 | >0.303 | >9.078 | 0.0242 (0.01972–0.02872) | >1416 |
Filament Ejection | EC50 | >0.303 | >9.078 | >40.939 | 60.055 (6.46–113.65) |
Polyp Debilitation | EC50 | >0.303 | <3.087 | 0.0258 (−0.00013–0.05176) | >1416 |
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Brefeld, D.; Di Mauro, V.; Kellermann, M.Y.; Nietzer, S.; Moeller, M.; Lütjens, L.H.; Pawlowski, S.; Petersen-Thiery, M.; Schupp, P.J. Acute Toxicity Assays with Adult Coral Fragments: A Method for Standardization. Toxics 2024, 12, 1. https://doi.org/10.3390/toxics12010001
Brefeld D, Di Mauro V, Kellermann MY, Nietzer S, Moeller M, Lütjens LH, Pawlowski S, Petersen-Thiery M, Schupp PJ. Acute Toxicity Assays with Adult Coral Fragments: A Method for Standardization. Toxics. 2024; 12(1):1. https://doi.org/10.3390/toxics12010001
Chicago/Turabian StyleBrefeld, David, Valentina Di Mauro, Matthias Y. Kellermann, Samuel Nietzer, Mareen Moeller, Laura H. Lütjens, Sascha Pawlowski, Mechtild Petersen-Thiery, and Peter J. Schupp. 2024. "Acute Toxicity Assays with Adult Coral Fragments: A Method for Standardization" Toxics 12, no. 1: 1. https://doi.org/10.3390/toxics12010001