Individual and Binary Mixture Toxicity of Five Nanoparticles in Marine Microalga Heterosigma akashiwo
Abstract
:1. Introduction
2. Results
2.1. First Stage of the Bioassay: Single Nanoparticle Assessment
2.2. Second Stage of the Bioassay: Binary Mixture Assessment
2.3. Particle Transformation Assessment
2.4. Summary
3. Discussion
4. Materials and Methods
4.1. Nanoparticles
4.2. Microalga Culture
4.3. Exposure Protocol
4.4. Cell Count, Staining, and Measurement Protocols
4.5. The Estimation of Particle Size Distribution and Particle Size Changes after Exposure
4.6. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Conflicts of Interest
References
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Effective Concentration | CdS | ZnS | TiO2 | SMB3 | SMB24 |
---|---|---|---|---|---|
EC50 | 21.33 (18.87–23.45) | 94.1 (87.4–100.7) | 141.7 (133.5–151.5) | 252.8 (210.4–352.2) | 3.6 (3.1–4.1) |
EC10 | 12.01 (11.67–12.35) | 53.1 (43.7–63.4) | 79.7 (72.4–88.4) | 143.6 (125.9–180.1) | 2.1 (1.9–2.2) |
Sample | Concentration (mg/L) | ||||
---|---|---|---|---|---|
CdS | ZnS | TiO2 | SMB3 | SMB24 | |
CdS | 12.0 | 0 | 0 | 0 | 0 |
ZnS | 0 | 53.0 | 0 | 0 | 0 |
TiO2 | 0 | 0 | 79.5 | 0 | 0 |
SMB3 | 0 | 0 | 0 | 143.5 | 0 |
SMB24 | 0 | 0 | 0 | 0 | 2.1 |
CdS + ZnS | 12.0 | 53.0 | 0 | 0 | 0 |
CdS + TiO2 | 12.0 | 0 | 79.5 | 0 | 0 |
CdS + SMB3 | 12.0 | 0 | 0 | 143.5 | 0 |
CdS + SMB24 | 12.0 | 0 | 0 | 0 | 2.1 |
ZnS + TiO2 | 0 | 53.0 | 79.5 | 0 | 0 |
ZnS + SMB3 | 0 | 53.0 | 0 | 143.5 | 0 |
ZnS + SMB24 | 0 | 53.0 | 0 | 0 | 2.1 |
TiO2 + SMB3 | 0 | 0 | 79.5 | 143.5 | 0 |
TiO2 + SMB24 | 0 | 0 | 79.5 | 0 | 2.1 |
SMB3 + SMB24 | 0 | 0 | 0 | 143.5 | 2.1 |
Sample | Microalgae Bioassay | Factors Affecting the Particle Agglomeration | ||||||
---|---|---|---|---|---|---|---|---|
EC50 (mg/L) or Combined Effect | Esterase Activity | Membrane Potential | ROS Generation | Cell Size | Time (96 h) | Combination | Alga | |
CdS | 21.33 | − | − | − | ns | −+ | na | + |
ZnS | 94.1 | ns | ns | ++ | − | − | na | + |
TiO2 | 141.7 | ns | ns | − | − | ++− | na | + |
SMB3 | 252.8 | ns | ns | ns | − | − | na | ++ |
SMB24 | 3.6 | − | ns | − | −+ | −+ | na | +− |
CdS + ZnS | ant | − | − | − | − | − | − | + |
CdS + TiO2 | syn | ns | − | − | + | ++− | − | −+ |
CdS + SMB3 | syn | − | − | − | ns | − | − | ++ |
CdS + SMB24 | ant | − | − | − | ns | − | − | + |
ZnS + TiO2 | syn | ns | ns | ns | − | ++− | ++− | + |
ZnS + SMB3 | syn | − | ns | ns | − | − | − | ++ |
ZnS + SMB24 | ant | ns | ns | ++ | − | − | − | + |
TiO2 + SMB3 | syn | ns | ns | − | ns | −+ | −+ | + |
TiO2 + SMB24 | add | + | ns | − | ++ | −+ | − | + |
SMB3 + SMB24 | add | − | ns | − | ns | − | − | ++ |
Sample | Average Diameter (nm) | Physical Properties | Impurities (%) | Structure Features | Synthesis or Manufacturer Information |
---|---|---|---|---|---|
CdS | 5–9 | band gap: 3.1 eV | – | Cubic crystal phase, sphere-like particles | [60] |
ZnS | 2.6–5.6 | band gap: 4.0 eV | – | Cubic crystal phase, sphere-like particles | [60] |
TiO2 | 32 | BET surface area: 45 m2/g | >99.9% TiO2 | Nanopowder, anatase crystal structure | Thermo Fisher GmbH, Kandel, Germany, CAS number 1317-70-0, product number 39953 |
SMB3 | 100 | Pore diameter: 3.45 nm; BET surface area: 1260 m2/g | >99.9% SiO2 | Mesoporous silicon dioxide, 3D structure | SMBTM 3 Property, CENNANO Co., Ltd., Ulsan, Korea |
SMB24 | 100 | Pore diameter: 2.72 nm; BET surface area: 1124 m2/g | ZnO and Ag | Mesoporous silicon dioxide with ZnO and Ag encapsulation, 3D structure | SMBTM 24 Property, CENNANO Co., Ltd., Ulsan, Korea |
Parameters | Conditions |
---|---|
Temperature | 20 ± 2 °C |
pH | 8.0 ± 0.2 |
Salinity | 33 ± 1‰ |
Light intensity | 300 μmol∙m-2∙s-1, cool white fluorescent |
Light cycle | 12:12 h light:dark |
Cultivation chamber | 250 mL Erlenmeyer flask |
Age of test organisms | 14–20 d, exponential growth phase |
Initial bioassay cell density | 1.2–1.4 × 104 cells mL−1 |
Control/diluent water | f/2 medium/0.22 µm filtered seawater |
Endpoint | Fluorescent Dye or Registered Parameter | Dye Concentration/Duration of Staining * | Instrument | Excitation Source (nm) | Emission Channel/ Band Width (nm) |
---|---|---|---|---|---|
Growth inhibition | PI | 15 µM/10 min | CytoFLEX | 488 | 610/20 |
Size | Forward scatter intensity (size calibration kit F13838 by Molecular Probes, USA) | – | CytoFLEX | 488 | FSC |
Esterase activity | FDA | 50 µM/30 min | GloMax | 490 | 510–570 |
Membrane potential | DiOC6 | 7.5 µM/10 min | GloMax | 490 | 510–570 |
ROS generation | H2DCFDA | 250 µM/40 min | GloMax | 490 | 510–570 |
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Pikula, K.; Johari, S.A.; Santos-Oliveira, R.; Golokhvast, K. Individual and Binary Mixture Toxicity of Five Nanoparticles in Marine Microalga Heterosigma akashiwo. Int. J. Mol. Sci. 2022, 23, 990. https://doi.org/10.3390/ijms23020990
Pikula K, Johari SA, Santos-Oliveira R, Golokhvast K. Individual and Binary Mixture Toxicity of Five Nanoparticles in Marine Microalga Heterosigma akashiwo. International Journal of Molecular Sciences. 2022; 23(2):990. https://doi.org/10.3390/ijms23020990
Chicago/Turabian StylePikula, Konstantin, Seyed Ali Johari, Ralph Santos-Oliveira, and Kirill Golokhvast. 2022. "Individual and Binary Mixture Toxicity of Five Nanoparticles in Marine Microalga Heterosigma akashiwo" International Journal of Molecular Sciences 23, no. 2: 990. https://doi.org/10.3390/ijms23020990