Exploring the Toxicity of Oxytetracycline in Earthworms (Eisenia fetida) Based on the Integrated Biomarker Response Method
Abstract
:1. Introduction
2. Materials and Methods
2.1. Experimental Materials
2.2. Experimental Design
2.3. Sample Collection
2.4. Determination of Enzyme Content and Activity
2.5. Biomarker Response Index (BRI)
2.6. Integrated Biomarker Response (IBR) Index
- (1)
- Homogenization:
- (2)
- Assignment: if the biomarkers of the treatment group are activated and compared with the control group, then Zi = Yi; otherwise, Zi = −Yi.
- (3)
- IBR star chart
2.7. Data Analyses
2.7.1. TOPSIS Model Construction
2.7.2. Path Analysis Model Construction
3. Results
3.1. Results of Simple Statistical Analysis
3.2. BRI of Various Oxidative Stress Indicators
3.3. Comprehensive Evaluation of the IBR
3.4. Path Analysis of Physiological Metabolism of Earthworms
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Appendix A. Statistical Data (BRt and BRc) of Indicators of Oxidative Stress Effects in Earthworms (Mean ± Standard Deviation)
Groups | Exposure Concentration (mg/kg) | Exposure Time (day) | Head of Earthwom | Tail of Earthworm | ||||||||||
CAT (U/mg prot) | SOD (U/mg prot) | POD (U/mg prot) | MDA (nmol/mg prot) | GST (U/mg prot) | GPX (U/mg prot) | CAT (U/mg prot) | SOD (U/mg prot) | POD (U/mg prot) | MDA (nmol/mg prot) | GST (U/mg prot) | GPX (U/mg prot) | |||
Short term | 0.00 | 1 | 69.19 ± 0.068 | 4.59 ± 0.022 | 2.71 ± 0.064 | 0.43 ± 0.002 | 6.46 ± 0.045 | 1.86 ± 0.016 | 55.41 ± 0.019 | 3.4 ± 0.019 | 1.82 ± 0.018 | 0.24 ± 0.014 | 4.35 ± 0.011 | 2.99 ± 0.015 |
2 | 62.62 ± 0.070 | 4.98 ± 0.002 | 2.51 ± 0.001 | 0.36 ± 0.004 | 4.76 ± 0.019 | 1.06 ± 0.020 | 55.55 ± 0.003 | 3.34 ± 0.006 | 1.42 ± 0.009 | 0.23 ± 0.004 | 7.52 ± 0.028 | 3.09 ± 0.023 | ||
3 | 74.23 ± 0.030 | 3.76 ± 0.023 | 2.49 ± 0.038 | 0.35 ± 0.029 | 8.16 ± 0.057 | 1.40 ± 0.025 | 53.14 ± 0.060 | 3.39 ± 0.034 | 1.22 ± 0.002 | 0.22 ± 0.006 | 6.14 ± 0.039 | 2.73 ± 0.020 | ||
4 | 72.86 ± 0.028 | 4.45 ± 0.043 | 2.46 ± 0.018 | 0.31 ± 0.005 | 8.14 ± 0.021 | 1.45 ± 0.029 | 58.92 ± 0.036 | 3.20 ± 0.036 | 1.18 ± 0.035 | 0.22 ± 0.028 | 6.27 ± 0.021 | 2.6 ± 0.039 | ||
5 | 66.57 ± 0.083 | 4.38 ± 0.037 | 2.47 ± 0.024 | 0.49 ± 0.026 | 11.38 ± 0.004 | 1.49 ± 0.006 | 56.46 ± 0.031 | 3.34 ± 0.061 | 1.48 ± 0.015 | 0.21 ± 0.034 | 7.63 ± 0.004 | 2.95 ± 0.014 | ||
6 | 58.68 ± 0.061 | 3.38 ± 0.025 | 2.20 ± 0.039 | 0.37 ± 0.039 | 6.12 ± 0.072 | 1.42 ± 0.050 | 59.63 ± 0.021 | 2.80 ± 0.036 | 1.13 ± 0.036 | 0.18 ± 0.034 | 7.33 ± 0.003 | 2.85 ± 0.020 | ||
7 | 62.28 ± 0.052 | 3.39 ± 0.037 | 1.81 ± 0.038 | 0.52 ± 0.010 | 9.08 ± 0.015 | 2.39 ± 0.039 | 60.63 ± 0.038 | 3.31 ± 0.076 | 1.17 ± 0.041 | 0.17 ± 0.008 | 5.75 ± 0.012 | 3.15 ± 0.016 | ||
8 | 76.25 ± 0.011 | 4.28 ± 0.001 | 1.77 ± 0.007 | 0.52 ± 0.010 | 9.36 ± 0.024 | 3.19 ± 0.026 | 53.39 ± 0.028 | 3.23 ± 0.093 | 1.07 ± 0.061 | 0.17 ± 0.037 | 7.90 ± 0.002 | 3.26 ± 0.055 | ||
9 | 75.56 ± 0.002 | 4.21 ± 0.064 | 2.25 ± 0.067 | 0.31 ± 0.019 | 9.16 ± 0.002 | 2.41 ± 0.055 | 58.18 ± 0.113 | 3.30 ± 0.016 | 1.21 ± 0.034 | 0.15 ± 0.007 | 6.72 ± 0.024 | 2.50 ± 0.029 | ||
10 | 64.95 ± 0.030 | 3.86 ± 0.068 | 2.10 ± 0.028 | 0.21 ± 0.016 | 5.19 ± 0.024 | 2.48 ± 0.029 | 59.07 ± 0.008 | 3.25 ± 0.058 | 1.35 ± 0.063 | 0.17 ± 0.019 | 6.73 ± 0.011 | 2.94 ± 0.015 | ||
0.036 | 1 | 82.41 ± 0.567 | 4.91 ± 0.445 | 3.27 ± 0.164 | 0.52 ± 0.285 | 7.13 ± 0.297 | 2.31 ± 0.021 | 63.24 ± 0.214 | 3.7 ± 0.126 | 2.1 ± 0.161 | 0.28 ± 0.254 | 4.69 ± 0.256 | 3.62 ± 0.045 | |
2 | 74.38 ± 0.006 | 5.31 ± 0.047 | 3.05 ± 0.172 | 0.44 ± 0.246 | 5.27 ± 0.277 | 1.3 ± 0.016 | 63.31 ± 0.39 | 3.63 ± 0.027 | 1.64 ± 0.252 | 0.27 ± 0.353 | 8.08 ± 0.247 | 3.76 ± 0.015 | ||
3 | 87.91 ± 0.311 | 4.02 ± 0.012 | 3.17 ± 0.16 | 0.42 ± 0.246 | 9.09 ± 0.268 | 1.73 ± 0.018 | 60.3 ± 0.456 | 3.75 ± 0.5 | 1.43 ± 0.228 | 0.25 ± 0.27 | 6.65 ± 0.3 | 3.33 ± 0.013 | ||
4 | 85.54 ± 0.015 | 4.78 ± 0.023 | 3.03 ± 0.153 | 0.37 ± 0.249 | 9.08 ± 0.289 | 1.8 ± 0.025 | 66.52 ± 0.351 | 3.53 ± 0.065 | 1.38 ± 0.19 | 0.25 ± 0.333 | 6.82 ± 0.279 | 3.17 ± 0.013 | ||
5 | 77.43 ± 0.563 | 4.75 ± 0.063 | 3.03 ± 0.161 | 0.6 ± 0.262 | 12.73 ± 0.283 | 1.86 ± 0.032 | 63.61 ± 0.005 | 3.59 ± 0.45 | 1.73 ± 0.228 | 0.24 ± 0.311 | 8.36 ± 0.286 | 3.62 ± 0.058 | ||
6 | 67.82 ± 0.727 | 3.64 ± 0.14 | 2.73 ± 0.166 | 0.45 ± 0.29 | 6.87 ± 0.249 | 1.77 ± 0.02 | 67.08 ± 0.295 | 3.11 ± 0.128 | 1.33 ± 0.211 | 0.21 ± 0.306 | 8.05 ± 0.166 | 3.51 ± 0.047 | ||
7 | 71.88 ± 0.578 | 3.63 ± 0.431 | 2.26 ± 0.174 | 0.64 ± 0.279 | 10.18 ± 0.255 | 3.02 ± 0.005 | 67.56 ± 0.015 | 3.6 ± 0.069 | 1.38 ± 0.191 | 0.2 ± 0.304 | 6.36 ± 0.215 | 3.89 ± 0.045 | ||
8 | 87.48 ± 0.683 | 4.62 ± 0.022 | 2.24 ± 0.126 | 0.64 ± 0.204 | 10.54 ± 0.172 | 4.06 ± 0.034 | 59.34 ± 0.534 | 3.53 ± 0.444 | 1.26 ± 0.201 | 0.2 ± 0.302 | 8.76 ± 0.18 | 4.04 ± 0.037 | ||
9 | 86.1 ± 0.365 | 4.57 ± 0.025 | 2.92 ± 0.138 | 0.38 ± 0.226 | 10.39 ± 0.287 | 3.06 ± 0.002 | 65.01 ± 0.721 | 3.66 ± 0.047 | 1.44 ± 0.239 | 0.17 ± 0.293 | 7.42 ± 0.244 | 3.11 ± 0.035 | ||
10 | 72.88 ± 0.516 | 4.21 ± 0.442 | 2.68 ± 0.136 | 0.25 ± 0.253 | 5.87 ± 0.281 | 3.17 ± 0.006 | 63.04 ± 0.566 | 3.61 ± 0.012 | 1.27 ± 0.194 | 0.2 ± 0.303 | 7.46 ± 0.257 | 3.67 ± 0.009 | ||
0.36 | 1 | 86.51 ± 0.379 | 5.66 ± 0.066 | 3.17 ± 0.165 | 0.52 ± 0.293 | 7.39 ± 0.255 | 2.17 ± 0.023 | 67.57 ± 0.645 | 4.08 ± 0.024 | 2 ± 0.171 | 0.28 ± 0.237 | 4.8 ± 0.292 | 3.39 ± 0.021 | |
2 | 78.01 ± 0.238 | 6.1 ± 0.492 | 2.95 ± 0.201 | 0.42 ± 0.281 | 5.46 ± 0.27 | 1.23 ± 0.031 | 67.68 ± 0.618 | 3.96 ± 0.057 | 1.57 ± 0.19 | 0.27 ± 0.225 | 8.33 ± 0.303 | 3.52 ± 0.026 | ||
3 | 92.14 ± 0.42 | 4.67 ± 0.042 | 2.99 ± 0.183 | 0.4 ± 0.286 | 9.41 ± 0.293 | 1.64 ± 0.017 | 64.56 ± 0.417 | 4.05 ± 0.013 | 1.36 ± 0.166 | 0.25 ± 0.251 | 6.82 ± 0.29 | 3.11 ± 0.024 | ||
4 | 90.1 ± 0.434 | 5.54 ± 0.012 | 2.88 ± 0.168 | 0.37 ± 0.289 | 9.42 ± 0.246 | 1.71 ± 0.022 | 71 ± 0.232 | 3.87 ± 0.025 | 1.32 ± 0.161 | 0.25 ± 0.232 | 6.98 ± 0.308 | 2.98 ± 0.018 | ||
5 | 81.65 ± 0.377 | 5.48 ± 0.024 | 2.92 ± 0.185 | 0.58 ± 0.243 | 13.16 ± 0.224 | 1.77 ± 0.007 | 67.59 ± 0.451 | 4.03 ± 0.068 | 1.66 ± 0.17 | 0.24 ± 0.258 | 8.52 ± 0.302 | 3.4 ± 0.022 | ||
6 | 71.78 ± 0.005 | 4.24 ± 0.063 | 2.64 ± 0.166 | 0.44 ± 0.317 | 7.14 ± 0.28 | 1.68 ± 0.007 | 71.27 ± 0.445 | 3.37 ± 0.148 | 1.27 ± 0.165 | 0.2 ± 0.257 | 8.21 ± 0.316 | 3.29 ± 0.013 | ||
7 | 75.84 ± 0.277 | 4.27 ± 0.141 | 2.15 ± 0.149 | 0.62 ± 0.26 | 10.61 ± 0.252 | 2.85 ± 0.068 | 72.33 ± 0.388 | 4.03 ± 0.491 | 1.33 ± 0.176 | 0.19 ± 0.278 | 6.46 ± 0.29 | 3.65 ± 0.045 | ||
8 | 91.9 ± 0.016 | 5.39 ± 0.491 | 2.08 ± 0.176 | 0.61 ± 0.288 | 10.98 ± 0.274 | 3.8 ± 0.085 | 63.53 ± 0.347 | 3.94 ± 0.952 | 1.21 ± 0.187 | 0.19 ± 0.274 | 8.92 ± 0.285 | 3.8 ± 0.023 | ||
9 | 90.77 ± 0.542 | 5.32 ± 0.025 | 2.61 ± 0.207 | 0.36 ± 0.294 | 10.78 ± 0.31 | 2.87 ± 0.05 | 68.98 ± 0.52 | 4.06 ± 0.032 | 1.38 ± 0.179 | 0.17 ± 0.258 | 7.63 ± 0.278 | 2.92 ± 0.025 | ||
10 | 77.6 ± 0.741 | 4.9 ± 0.028 | 2.47 ± 0.194 | 0.24 ± 0.295 | 6.12 ± 0.291 | 2.96 ± 0.031 | 70.02 ± 0.385 | 3.99 ± 0.514 | 1.54 ± 0.169 | 0.19 ± 0.281 | 7.66 ± 0.274 | 3.43 ± 0.009 | ||
3.6 | 1 | 94.66 ± 0.311 | 6.86 ± 0.491 | 3.31 ± 0.158 | 0.5 ± 0.238 | 7.7 ± 0.265 | 2.1 ± 0.022 | 72.34 ± 0.218 | 4.56 ± 0.065 | 2.13 ± 0.161 | 0.27 ± 0.266 | 5 ± 0.305 | 3.28 ± 0.042 | |
2 | 85.55 ± 0.454 | 7.38 ± 0.066 | 3.06 ± 0.16 | 0.42 ± 0.28 | 5.7 ± 0.271 | 1.19 ± 0.028 | 70.89 ± 0.485 | 4.46 ± 0.413 | 1.67 ± 0.162 | 0.26 ± 0.276 | 8.68 ± 0.321 | 3.4 ± 0.037 | ||
3 | 100.39 ± 0.318 | 5.61 ± 0.448 | 3.09 ± 0.138 | 0.4 ± 0.217 | 9.8 ± 0.286 | 1.58 ± 0.032 | 67.51 ± 0.528 | 4.42 ± 0.117 | 1.45 ± 0.16 | 0.24 ± 0.264 | 7.14 ± 0.285 | 3.01 ± 0.028 | ||
4 | 98.08 ± 0.216 | 6.84 ± 0.133 | 3.08 ± 0.167 | 0.36 ± 0.261 | 9.8 ± 0.292 | 1.65 ± 0.014 | 74.64 ± 0.396 | 4.22 ± 0.026 | 1.4 ± 0.166 | 0.24 ± 0.237 | 7.3 ± 0.308 | 2.88 ± 0.022 | ||
5 | 89.67 ± 0.362 | 6.54 ± 0.135 | 3.11 ± 0.154 | 0.56 ± 0.244 | 13.77 ± 0.293 | 1.69 ± 0.017 | 71.35 ± 0.006 | 4.67 ± 0.46 | 1.76 ± 0.16 | 0.23 ± 0.312 | 8.95 ± 0.311 | 3.29 ± 0.008 | ||
6 | 78.1 ± 0.392 | 5.12 ± 0.045 | 2.77 ± 0.152 | 0.44 ± 0.243 | 7.45 ± 0.167 | 1.6 ± 0.041 | 75.33 ± 0.334 | 3.75 ± 0.062 | 1.35 ± 0.164 | 0.2 ± 0.273 | 8.62 ± 0.261 | 3.19 ± 0.006 | ||
7 | 82.77 ± 0.301 | 5.21 ± 0.012 | 2.26 ± 0.191 | 0.59 ± 0.306 | 11.08 ± 0.314 | 2.73 ± 0.033 | 76.39 ± 0.017 | 4.45 ± 0.4 | 1.4 ± 0.156 | 0.18 ± 0.26 | 6.82 ± 0.305 | 3.52 ± 0.028 | ||
8 | 100.47 ± 0.442 | 6.48 ± 0.023 | 2.17 ± 0.157 | 0.58 ± 0.268 | 11.42 ± 0.307 | 3.64 ± 0.045 | 66.97 ± 0.588 | 4.33 ± 0.119 | 1.28 ± 0.181 | 0.18 ± 0.282 | 9.36 ± 0.176 | 3.67 ± 0.007 | ||
9 | 98.87 ± 0.379 | 6.47 ± 0.061 | 2.79 ± 0.161 | 0.34 ± 0.298 | 11.22 ± 0.305 | 2.77 ± 0.033 | 73.06 ± 0.37 | 4.45 ± 1.413 | 1.46 ± 0.171 | 0.16 ± 0.274 | 8.01 ± 0.268 | 2.82 ± 0.042 | ||
10 | 84.83 ± 0.218 | 5.88 ± 0.139 | 2.59 ± 0.177 | 0.23 ± 0.293 | 6.37 ± 0.295 | 2.86 ± 0.018 | 74.2 ± 0.558 | 4.44 ± 0.045 | 1.63 ± 0.194 | 0.19 ± 0.275 | 8.07 ± 0.292 | 3.32 ± 0.017 | ||
36 | 1 | 106.99 ± 0.184 | 7.78 ± 0.037 | 3.47 ± 0.032 | 0.56 ± 0.028 | 8.61 ± 0.316 | 2.52 ± 0.032 | 80.2 ± 0.387 | 5.43 ± 0.012 | 2.2 ± 0.16 | 0.31 ± 0.278 | 5.55 ± 0.283 | 3.84 ± 0.051 | |
2 | 96.08 ± 0.123 | 8.46 ± 0.066 | 3.19 ± 0.031 | 0.47 ± 0.031 | 6.34 ± 0.298 | 1.43 ± 0.046 | 80.31 ± 0.192 | 5.27 ± 0.023 | 1.72 ± 0.18 | 0.29 ± 0.218 | 9.62 ± 0.28 | 3.97 ± 0.032 | ||
3 | 113.36 ± 0.444 | 6.42 ± 0.015 | 3.14 ± 0.032 | 0.46 ± 0.029 | 10.9 ± 0.34 | 1.9 ± 0.061 | 76.41 ± 0.364 | 5.36 ± 0.068 | 1.49 ± 0.16 | 0.28 ± 0.236 | 7.87 ± 0.298 | 3.52 ± 0.008 | ||
4 | 109.37 ± 0.021 | 7.61 ± 0.161 | 3.13 ± 0.03 | 0.4 ± 0.027 | 10.93 ± 0.268 | 1.97 ± 0.033 | 84.69 ± 0.385 | 5.08 ± 0.139 | 1.44 ± 0.172 | 0.28 ± 0.25 | 8.06 ± 0.27 | 3.37 ± 0.02 | ||
5 | 99.73 ± 0.012 | 7.51 ± 0.028 | 3.16 ± 0.031 | 0.64 ± 0.028 | 15.33 ± 0.267 | 2.04 ± 0.034 | 81.06 ± 0.346 | 5.31 ± 0.449 | 1.81 ± 0.16 | 0.27 ± 0.226 | 9.86 ± 0.258 | 3.81 ± 0.031 | ||
6 | 87.18 ± 0.338 | 5.81 ± 0.009 | 2.79 ± 0.033 | 0.49 ± 0.029 | 8.27 ± 0.289 | 1.94 ± 0.036 | 85.14 ± 0.005 | 4.47 ± 0.861 | 1.39 ± 0.161 | 0.23 ± 0.259 | 9.52 ± 0.304 | 3.69 ± 0.011 | ||
7 | 91.8 ± 0.124 | 5.85 ± 0.074 | 2.29 ± 0.03 | 0.69 ± 0.027 | 12.34 ± 0.285 | 3.28 ± 0.042 | 86.31 ± 0.286 | 5.29 ± 0.026 | 1.44 ± 0.158 | 0.22 ± 0.255 | 7.52 ± 0.242 | 4.09 ± 0.027 | ||
8 | 111.86 ± 0.102 | 7.39 ± 0.078 | 2.29 ± 0.033 | 0.68 ± 0.03 | 12.76 ± 0.286 | 4.4 ± 0.025 | 76.73 ± 0.015 | 5.18 ± 0.51 | 1.33 ± 0.157 | 0.22 ± 0.265 | 10.32 ± 0.218 | 4.27 ± 0.043 | ||
9 | 110.11 ± 0.319 | 7.25 ± 0.02 | 2.93 ± 0.031 | 0.41 ± 0.029 | 12.56 ± 0.301 | 3.34 ± 0.04 | 83.72 ± 0.482 | 5.34 ± 0.065 | 1.51 ± 0.18 | 0.19 ± 0.261 | 8.79 ± 0.28 | 3.29 ± 0.05 | ||
10 | 93.89 ± 0.071 | 6.67 ± 0.003 | 2.75 ± 0.031 | 0.27 ± 0.029 | 7.16 ± 0.326 | 3.45 ± 0.028 | 84.62 ± 0.722 | 5.26 ± 0.455 | 1.68 ± 0.184 | 0.22 ± 0.272 | 8.85 ± 0.248 | 3.86 ± 0.026 | ||
72 | 1 | 103.01 ± 0.383 | 6.79 ± 0.43 | 3.79 ± 0.156 | 0.61 ± 0.242 | 9.36 ± 0.299 | 2.77 ± 0.028 | 73.06 ± 0.225 | 4.8 ± 0.038 | 2.39 ± 0.187 | 0.32 ± 0.289 | 5.95 ± 0.241 | 4.18 ± 0.035 | |
2 | 92.81 ± 0.342 | 7.37 ± 0.721 | 3.47 ± 0.122 | 0.5 ± 0.211 | 6.88 ± 0.309 | 1.57 ± 0.03 | 73.04 ± 0.351 | 4.68 ± 0.189 | 1.87 ± 0.184 | 0.31 ± 0.284 | 10.31 ± 0.24 | 4.33 ± 0.06 | ||
3 | 109.7 ± 0.336 | 5.6 ± 0.027 | 3.43 ± 0.148 | 0.49 ± 0.274 | 11.84 ± 0.281 | 2.09 ± 0.009 | 69.82 ± 0.321 | 4.74 ± 0.077 | 1.62 ± 0.182 | 0.29 ± 0.331 | 8.43 ± 0.277 | 3.84 ± 0.029 | ||
4 | 107.01 ± 0.005 | 6.67 ± 0.044 | 3.41 ± 0.155 | 0.43 ± 0.247 | 11.84 ± 0.301 | 2.16 ± 0.033 | 77.03 ± 0.505 | 4.54 ± 0.074 | 1.57 ± 0.219 | 0.29 ± 0.327 | 8.64 ± 0.262 | 3.68 ± 0.048 | ||
5 | 97 ± 0.263 | 6.61 ± 0.01 | 3.48 ± 0.151 | 0.69 ± 0.239 | 16.57 ± 0.268 | 2.23 ± 0.016 | 73.78 ± 0.547 | 4.72 ± 0.399 | 1.97 ± 0.208 | 0.28 ± 0.338 | 10.55 ± 0.25 | 4.18 ± 0.039 | ||
6 | 84.97 ± 0.014 | 5.11 ± 0.022 | 3.13 ± 0.161 | 0.52 ± 0.265 | 8.96 ± 0.277 | 2.12 ± 0.011 | 77.57 ± 0.388 | 3.99 ± 0.439 | 1.51 ± 0.21 | 0.24 ± 0.327 | 10.15 ± 0.171 | 4.06 ± 0.035 | ||
7 | 89.32 ± 0.491 | 5.14 ± 0.062 | 2.6 ± 0.146 | 0.74 ± 0.318 | 13.34 ± 0.291 | 3.6 ± 0.048 | 78.59 ± 0.481 | 4.74 ± 0.332 | 1.56 ± 0.219 | 0.23 ± 0.319 | 7.96 ± 0.241 | 4.48 ± 0.052 | ||
8 | 108.71 ± 0.747 | 6.49 ± 0.142 | 2.54 ± 0.18 | 0.73 ± 0.287 | 13.76 ± 0.175 | 4.83 ± 0.026 | 68.91 ± 0.169 | 4.65 ± 0.549 | 1.43 ± 0.214 | 0.23 ± 0.309 | 11 ± 0.307 | 4.66 ± 0.028 | ||
9 | 106.57 ± 0.5 | 6.43 ± 0.395 | 3.19 ± 0.173 | 0.44 ± 0.295 | 13.54 ± 0.304 | 3.66 ± 0.035 | 75.23 ± 0.532 | 4.77 ± 0.061 | 1.63 ± 0.193 | 0.2 ± 0.341 | 9.38 ± 0.263 | 3.6 ± 0.055 | ||
10 | 91.11 ± 0.592 | 5.89 ± 0.81 | 3.04 ± 0.185 | 0.29 ± 0.256 | 7.65 ± 0.296 | 3.78 ± 0.032 | 76.03 ± 0.629 | 4.69 ± 0.502 | 1.82 ± 0.21 | 0.23 ± 0.324 | 9.42 ± 0.289 | 4.24 ± 0.066 | ||
Long-term | 0.0 | 10 | 53.86 ± 0.053 | 3.92 ± 0.066 | 1.90 ± 0.026 | 0.23 ± 0.044 | 9.62 ± 0.028 | 1.56 ± 0.023 | 65.42 ± 0.014 | 3.56 ± 0.052 | 1.56 ± 0.014 | 0.15 ± 0.027 | 7.82 ± 0.004 | 2.7 ± 0.014 |
20 | 52.99 ± 0.020 | 4.60 ± 0.067 | 2.27 ± 0.038 | 0.41 ± 0.035 | 10.34 ± 0.039 | 3.30 ± 0.02 | 60.60 ± 0.006 | 2.85 ± 0.029 | 1.14 ± 0.022 | 0.20 ± 0.003 | 7.67 ± 0.003 | 2.12 ± 0.02 | ||
30 | 60.89 ± 0.043 | 4.17 ± 0.028 | 1.98 ± 0.019 | 0.45 ± 0.012 | 5.00 ± 0.021 | 6.67 ± 0.039 | 57.07 ± 0.067 | 3.13 ± 0.104 | 1.46 ± 0.038 | 0.21 ± 0.036 | 8.17 ± 0.012 | 2.12 ± 0.016 | ||
0.036 | 10 | 60.44 ± 0.371 | 4.34 ± 0.044 | 2.44 ± 0.155 | 0.29 ± 0.292 | 11 ± 0.313 | 2.03 ± 0.023 | 71.48 ± 0.515 | 3.96 ± 0.058 | 1.87 ± 0.099 | 0.18 ± 0.241 | 8.73 ± 0.356 | 3.38 ± 0.037 | |
20 | 57.93 ± 0.166 | 5.23 ± 0.009 | 2.97 ± 0.103 | 0.51 ± 0.226 | 12.23 ± 0.268 | 4.36 ± 0.011 | 70.42 ± 0.215 | 3.24 ± 0.031 | 1.36 ± 0.111 | 0.24 ± 0.288 | 8.69 ± 0.333 | 2.72 ± 0.011 | ||
30 | 65.07 ± 0.305 | 4.91 ± 0.022 | 2.6 ± 0.116 | 0.56 ± 0.212 | 6.01 ± 0.302 | 8.93 ± 0.053 | 59.52 ± 0.365 | 3.6 ± 0.5 | 1.85 ± 0.15 | 0.25 ± 0.372 | 9.34 ± 0.517 | 2.79 ± 0.02 | ||
0.36 | 10 | 64.35 ± 0.368 | 4.93 ± 0.057 | 2.2 ± 0.143 | 0.27 ± 0.237 | 11.41 ± 0.252 | 1.88 ± 0.038 | 77.05 ± 0.012 | 4.41 ± 0.022 | 1.73 ± 0.133 | 0.17 ± 0.271 | 8.94 ± 0.253 | 3.15 ± 0.025 | |
20 | 59.93 ± 0.317 | 5.85 ± 0.12 | 2.67 ± 0.126 | 0.46 ± 0.249 | 12.61 ± 0.274 | 4.08 ± 0.032 | 70.1 ± 0.436 | 3.54 ± 0.411 | 1.3 ± 0.116 | 0.22 ± 0.288 | 9.01 ± 0.241 | 2.53 ± 0.033 | ||
30 | 65.31 ± 0.005 | 5.28 ± 0.413 | 2.3 ± 0.141 | 0.5 ± 0.233 | 6.2 ± 0.26 | 8.5 ± 0.019 | 64.1 ± 0.542 | 3.91 ± 0.052 | 1.66 ± 0.137 | 0.22 ± 0.241 | 9.78 ± 0.277 | 2.6 ± 0.032 | ||
3.6 | 10 | 70.35 ± 0.366 | 6.08 ± 0.041 | 2.39 ± 0.15 | 0.26 ± 0.353 | 11.8 ± 0.336 | 1.82 ± 0.027 | 81.54 ± 0.214 | 4.76 ± 0.027 | 1.86 ± 0.159 | 0.17 ± 0.246 | 9.25 ± 0.309 | 3.07 ± 0.011 | |
20 | 66.98 ± 0.17 | 6.97 ± 0.01 | 2.79 ± 0.098 | 0.45 ± 0.281 | 13.12 ± 0.326 | 3.95 ± 0.012 | 73.31 ± 0.389 | 3.85 ± 0.47 | 1.39 ± 0.193 | 0.22 ± 0.324 | 9.32 ± 0.467 | 2.47 ± 0.021 | ||
30 | 73.53 ± 0.312 | 6.48 ± 0.02 | 2.5 ± 0.112 | 0.48 ± 0.294 | 6.54 ± 0.361 | 8.19 ± 0.059 | 67.34 ± 0.412 | 4.34 ± 0.07 | 1.81 ± 0.21 | 0.22 ± 0.298 | 10.05 ± 0.306 | 2.53 ± 0.037 | ||
36 | 10 | 77.86 ± 0.487 | 6.72 ± 0.055 | 2.53 ± 0.136 | 0.31 ± 0.277 | 13.13 ± 0.258 | 2.19 ± 0.037 | 92.39 ± 0.363 | 5.88 ± 0.445 | 1.96 ± 0.176 | 0.2 ± 0.304 | 10.4 ± 0.256 | 3.58 ± 0.025 | |
20 | 73.32 ± 0.344 | 8.06 ± 0.115 | 3.02 ± 0.115 | 0.57 ± 0.304 | 14.4 ± 0.278 | 4.77 ± 0.038 | 83.48 ± 0.006 | 4.78 ± 0.132 | 1.48 ± 0.179 | 0.27 ± 0.287 | 10.39 ± 0.265 | 2.9 ± 0.058 | ||
30 | 80.84 ± 0.005 | 7.38 ± 0.395 | 2.68 ± 0.138 | 0.63 ± 0.3 | 7.05 ± 0.309 | 9.86 ± 0.023 | 76.54 ± 0.298 | 5.31 ± 0.133 | 1.92 ± 0.166 | 0.28 ± 0.248 | 11.34 ± 0.289 | 2.97 ± 0.033 | ||
72 | 10 | 75.56 ± 0.268 | 6.29 ± 0.021 | 2.77 ± 0.14 | 0.33 ± 0.284 | 14.22 ± 0.249 | 2.35 ± 0.037 | 85.01 ± 0.307 | 5.41 ± 0.506 | 2.11 ± 0.161 | 0.21 ± 0.278 | 11.09 ± 0.272 | 3.94 ± 0.055 | |
20 | 72.85 ± 0.011 | 7.53 ± 0.018 | 3.36 ± 0.117 | 0.59 ± 0.304 | 15.39 ± 0.311 | 5.09 ± 0.03 | 76.48 ± 0.448 | 4.42 ± 0.061 | 1.58 ± 0.179 | 0.28 ± 0.346 | 11.04 ± 0.296 | 3.2 ± 0.042 | ||
30 | 79.91 ± 0.411 | 6.9 ± 0.427 | 2.99 ± 0.113 | 0.65 ± 0.343 | 7.64 ± 0.366 | 10.49 ± 0.041 | 69.94 ± 0.358 | 4.96 ± 0.431 | 2.06 ± 0.171 | 0.3 ± 0.285 | 12 ± 0.264 | 3.26 ± 0.033 | ||
Abbreviations: CAT, catalase activity; SOD, superoxide dismutase activity; POD, peroxidase activity; MDA, malondialdehyde content; GST, glutathione S-transferase activity; GPX, glutathione peroxidase activity. The control group (0.0 mg/kg) data were the BRc; the experimental groups' (0.036, 0.36, 3.6, 36, and 72 mg/kg) data were the BRt. |
Appendix B. Assumption Test of Variance Analysis
Groups | Tissue | Exposure Time (day) | Indices | |||||||||||
CAT | SOD | POD | MDA | GST | GPX | |||||||||
Shapiro–Wilk | Sig. | Shapiro–Wilk | Sig. | Shapiro–Wilk | Sig. | Shapiro–Wilk | Sig. | Shapiro–Wilk | Sig. | Shapiro–Wilk | Sig. | |||
Short-term | Head | 1 | 0.936 | 0.641 | 0.958 | 0.797 | 0.896 | 0.388 | 0.864 | 0.244 | 0.929 | 0.587 | 0.941 | 0.674 |
2 | 0.938 | 0.655 | 0.958 | 0.797 | 0.896 | 0.387 | 0.884 | 0.326 | 0.925 | 0.562 | 0.940 | 0.665 | ||
3 | 0.936 | 0.636 | 0.954 | 0.766 | 0.913 | 0.488 | 0.916 | 0.506 | 0.922 | 0.543 | 0.938 | 0.649 | ||
4 | 0.927 | 0.575 | 0.956 | 0.778 | 0.941 | 0.673 | 0.909 | 0.461 | 0.922 | 0.543 | 0.938 | 0.652 | ||
5 | 0.934 | 0.625 | 0.957 | 0.787 | 0.919 | 0.526 | 0.939 | 0.658 | 0.922 | 0.541 | 0.948 | 0.721 | ||
6 | 0.936 | 0.639 | 0.962 | 0.819 | 0.824 | 0.126 | 0.888 | 0.347 | 0.931 | 0.606 | 0.948 | 0.726 | ||
7 | 0.938 | 0.651 | 0.951 | 0.744 | 0.818 | 0.112 | 0.967 | 0.853 | 0.932 | 0.608 | 0.955 | 0.773 | ||
8 | 0.935 | 0.63 | 0.956 | 0.778 | 0.934 | 0.621 | 0.971 | 0.884 | 0.927 | 0.576 | 0.955 | 0.773 | ||
9 | 0.945 | 0.703 | 0.954 | 0.762 | 0.972 | 0.888 | 0.958 | 0.793 | 0.920 | 0.530 | 0.944 | 0.696 | ||
10 | 0.946 | 0.707 | 0.948 | 0.720 | 0.951 | 0.745 | 0.954 | 0.768 | 0.925 | 0.563 | 0.946 | 0.705 | ||
Levene statistic Sig. | 1.000 | 1.000 | 0.999 | 0.998 | 1.000 | 1.000 | ||||||||
Tail | 1 | 0.984 | 0.955 | 0.987 | 0.969 | 0.912 | 0.479 | 0.898 | 0.399 | 0.912 | 0.479 | 0.957 | 0.79 | |
2 | 0.985 | 0.959 | 0.987 | 0.969 | 0.919 | 0.524 | 0.886 | 0.338 | 0.919 | 0.524 | 0.96 | 0.807 | ||
3 | 0.989 | 0.977 | 0.982 | 0.945 | 0.92 | 0.53 | 0.914 | 0.491 | 0.92 | 0.53 | 0.959 | 0.798 | ||
4 | 0.985 | 0.961 | 0.979 | 0.93 | 0.913 | 0.489 | 0.923 | 0.55 | 0.913 | 0.489 | 0.957 | 0.787 | ||
5 | 0.984 | 0.955 | 0.989 | 0.974 | 0.909 | 0.464 | 0.9 | 0.412 | 0.909 | 0.464 | 0.953 | 0.756 | ||
6 | 0.984 | 0.957 | 0.965 | 0.843 | 0.907 | 0.452 | 0.918 | 0.517 | 0.907 | 0.452 | 0.952 | 0.75 | ||
7 | 0.988 | 0.973 | 0.984 | 0.954 | 0.905 | 0.438 | 0.925 | 0.566 | 0.904 | 0.434 | 0.963 | 0.826 | ||
8 | 0.978 | 0.924 | 0.995 | 0.994 | 0.908 | 0.456 | 0.925 | 0.565 | 0.908 | 0.457 | 0.961 | 0.815 | ||
9 | 0.974 | 0.899 | 0.994 | 0.991 | 0.927 | 0.578 | 0.93 | 0.597 | 0.927 | 0.578 | 0.954 | 0.767 | ||
10 | 0.99 | 0.978 | 0.993 | 0.988 | 0.928 | 0.582 | 0.922 | 0.545 | 0.928 | 0.582 | 0.956 | 0.783 | ||
Levene statistic Sig. | 1.000 | 1.000 | 1.000 | 0.999 | 1.000 | 1.000 | ||||||||
Long-term | Head | 10 | 0.946 | 0.707 | 0.92 | 0.528 | 0.979 | 0.93 | 0.931 | 0.601 | 0.919 | 0.52 | 0.955 | 0.772 |
20 | 0.876 | 0.293 | 0.951 | 0.748 | 0.958 | 0.795 | 0.899 | 0.403 | 0.926 | 0.569 | 0.942 | 0.679 | ||
30 | 0.847 | 0.187 | 0.924 | 0.555 | 0.976 | 0.912 | 0.907 | 0.45 | 0.944 | 0.694 | 0.936 | 0.637 | ||
Levene statistic Sig. | 0.970 | 0.998 | 0.976 | 0.741 | 0.959 | 0.982 | ||||||||
Tail | 10 | 0.996 | 0.997 | 0.973 | 0.897 | 0.892 | 0.367 | 0.942 | 0.681 | 0.892 | 0.367 | 0.947 | 0.713 | |
20 | 0.881 | 0.315 | 0.96 | 0.809 | 0.918 | 0.519 | 0.922 | 0.543 | 0.918 | 0.519 | 0.934 | 0.624 | ||
30 | 0.993 | 0.99 | 0.956 | 0.779 | 0.915 | 0.498 | 0.918 | 0.516 | 0.915 | 0.498 | 0.946 | 0.711 | ||
Levene statistic Sig. | 0.885 | 0.985 | 0.969 | 0.651 | 0.969 | 0.974 | ||||||||
Abbreviations: CAT, catalase activity; SOD, superoxide dismutase activity; POD, peroxidase activity; MDA, malondialdehyde content; GST, glutathione S-transferase activity; GPX, glutathione peroxidase activity. |
Appendix C. Analysis of Variance of Enzyme Activity in Earthworms under Oxytetracycline Stress
Groups | Tissue | Indices | Exposure Time | Exposure Concentration | ||||||||
III-Sums of Squares | df | Mean Square | F Value | Sig. | III-Sums of Squares | df | Mean Square | F Value | Sig. | |||
Short-term | Head | CAT | 276.291 | 9 | 30.699 | 71.575 | 0 | 8006.848 | 4 | 2001.712 | 4667.032 | 0 |
SOD | 88.5 | 9 | 9.833 | 12.906 | 0 | 24,969.136 | 4 | 6242.284 | 8192.754 | 0 | ||
POD | 67.193 | 9 | 7.466 | 2.075 | 0.059 | 2890.796 | 4 | 722.699 | 200.86 | 0 | ||
MDA | 18.582 | 9 | 2.065 | 0.805 | 0.614 | 4775.662 | 4 | 1193.915 | 465.762 | 0 | ||
GST | 64.359 | 9 | 7.151 | 66.613 | 0 | 8033.661 | 4 | 2008.415 | 18,708.515 | 0 | ||
GPX | 77.665 | 9 | 8.629 | 58.385 | 0 | 8727.362 | 4 | 2181.841 | 14,761.86 | 0 | ||
Tail | CAT | 52.925 | 9 | 5.881 | 9.017 | 0 | 5575.229 | 4 | 1393.807 | 2137.276 | 0 | |
SOD | 81.531 | 9 | 9.059 | 4.776 | 0 | 14,755.069 | 4 | 3688.767 | 1944.855 | 0 | ||
POD | 72.12 | 9 | 8.013 | 83.078 | 0 | 6009.643 | 4 | 1502.411 | 15,576.324 | 0 | ||
MDA | 1.421 | 9 | 0.158 | 0.173 | 0.996 | 4604.688 | 4 | 1151.172 | 1259.084 | 0 | ||
GST | 71.396 | 9 | 7.933 | 72.262 | 0 | 5992.903 | 4 | 1498.226 | 13,647.709 | 0 | ||
GPX | 63.105 | 9 | 7.012 | 85.863 | 0 | 5926.588 | 4 | 1481.647 | 18,143.678 | 0 | ||
Long-term | Head | CAT | 233.639 | 2 | 116.819 | 54.512 | 0 | 2074.226 | 4 | 518.557 | 241.975 | 0 |
SOD | 38.37 | 2 | 19.185 | 5.291 | 0.034 | 7733.931 | 4 | 1933.483 | 533.269 | 0 | ||
POD | 15.618 | 2 | 7.809 | 4.042 | 0.061 | 1666.204 | 4 | 416.551 | 215.579 | 0 | ||
MDA | 8.807 | 2 | 4.404 | 0.804 | 0.481 | 2827.165 | 4 | 706.791 | 128.969 | 0 | ||
GST | 81.381 | 2 | 40.69 | 51.058 | 0 | 2108.08 | 4 | 527.02 | 661.302 | 0 | ||
GPX | 111.788 | 2 | 55.894 | 118.74 | 0 | 2449.679 | 4 | 612.42 | 1301.009 | 0 | ||
Tail | CAT | 112.099 | 2 | 56.049 | 9.15 | 0.009 | 1369.05 | 4 | 342.263 | 55.874 | 0 | |
SOD | 51.341 | 2 | 25.671 | 22.514 | 0.001 | 5888.623 | 4 | 1472.156 | 1291.149 | 0 | ||
POD | 55.665 | 2 | 27.833 | 61.302 | 0 | 2082.227 | 4 | 520.557 | 1146.533 | 0 | ||
MDA | 4.862 | 2 | 2.431 | 0.399 | 0.684 | 2429.834 | 4 | 607.458 | 99.598 | 0 | ||
GST | 55.617 | 2 | 27.809 | 61.235 | 0 | 2082.586 | 4 | 520.646 | 1146.479 | 0 | ||
GPX | 105.175 | 2 | 52.587 | 121.865 | 0 | 2228.599 | 4 | 557.15 | 1291.128 | 0 | ||
Abbreviations: CAT, catalase activity; SOD, superoxide dismutase activity; POD, peroxidase activity; MDA, malondialdehyde content; GST, glutathione S-transferase activity; GPX, glutathione peroxidase activity. |
Appendix D. The Concentration of Oxytetracycline Concentration Detected by High-Performance Liquid Chromatography
Experimental Setting Concentration (mg/kg) | Oxytetracycline Concentration in Soil (mg/kg) | Oxytetracycline Concentration in Earthworm (mg/kg) |
0.036 | 0.03 | <0.01 |
0.36 | 0.32 | |
3.6 | 3.19 | |
36 | 32.58 | |
72 | 65.52 |
Appendix E. EC50 Data of Q
Groups | Exposure Time (day) | EC 50 of Q (mg/kg) | |
Head | Tail | ||
Short-term | 1 | 23.510 | 81.953 |
2 | 24.719 | 107.282 | |
3 | 24.561 | 124.815 | |
4 | 20.778 | 105.317 | |
5 | 22.753 | 76.976 | |
6 | 21.503 | 94.562 | |
7 | 20.330 | 92.127 | |
8 | 22.602 | 88.409 | |
9 | 22.420 | 75.166 | |
10 | 22.498 | 65.412 | |
Long-term | 10 | 22.983 | 67.360 |
20 | 22.620 | 66.252 | |
30 | 20.387 | 51.661 |
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Concentration Group (mg/kg) | Short-Term | Long-Term | ||
---|---|---|---|---|
Head | Tail | Head | Tail | |
0.036 | CAT **, SOD ** | CAT **, SOD ** | POD **, SOD ** | CAT **, SOD ** |
0.36 | SOD **, CAT ** | SOD **, POD ** | CAT **, SOD ** | POD **, SOD ** |
3.6 | SOD **, GPX ** | POD **, GST ** | SOD **, GPX ** | POD **, GST ** |
36 | MDA **, GPX ** | MDA *, GST ** | MDA *, GPX ** | GST **, MDA * |
72 | GPX **, MDA * | GST **, MDA * | GPX **, MDA * | GST **, MDA ** |
Concentration Group (mg/kg) | p Value | Durbin–Watson | Determination Coefficient | Residual Path Coefficient |
---|---|---|---|---|
0.036 | 0.0001 | 1.286 | 0.999 | 0.036 |
0.36 | 0.0001 | 2.866 | 0.986 | 0.117 |
3.6 | 0.0001 | 2.925 | 0.991 | 0.095 |
36 | 0.0001 | 2.656 | 1.000 | 0.010 |
72 | 0.0001 | 1.303 | 1.000 | 0.012 |
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Zhou, H.; Jiao, X.; Li, Y. Exploring the Toxicity of Oxytetracycline in Earthworms (Eisenia fetida) Based on the Integrated Biomarker Response Method. Toxics 2024, 12, 310. https://doi.org/10.3390/toxics12050310
Zhou H, Jiao X, Li Y. Exploring the Toxicity of Oxytetracycline in Earthworms (Eisenia fetida) Based on the Integrated Biomarker Response Method. Toxics. 2024; 12(5):310. https://doi.org/10.3390/toxics12050310
Chicago/Turabian StyleZhou, Haoran, Xiaoguang Jiao, and Yunfei Li. 2024. "Exploring the Toxicity of Oxytetracycline in Earthworms (Eisenia fetida) Based on the Integrated Biomarker Response Method" Toxics 12, no. 5: 310. https://doi.org/10.3390/toxics12050310