## 3 Results
### 1 h and 24 h Binary Mixture of Cd + ZnO--MixTOX Analysis
\begin{table}
\begin{tabular}{c c c c c c} \hline \hline \multicolumn{2}{c}{**CdCl\({}_{2}\) + ZnO Exposure**} & **Parameter** & \multicolumn{4}{c}{**The Concentration Addition (CA)-Based Module**} \\  & & **CA** & **S/A** & **DR** & **DL** \\ \hline
1 h & \(a\) & – & –5.325 & –3.654 & –15.34 \\  & \(b\) & – & – & –5.365 & –8.324 \\  & \(R^{2}\) & 0.47 & 0.97 & 0.98 & 0.99 \\  & \(p(\chi 2)\) CA vs. & – & 0.000 * & 0.000 * & 0.000 * \\  & S/A vs. & – & – & 7.4 \(\times\) 10\({}^{-11}\) * & 5.17 \(\times\) 10\({}^{-14}\) * \\
24 h & \(a\) & – & –0.043 & –2.805 & –0.205 \\  & \(b\) & – & – & 5.643 & 0.362 \\  & \(R^{2}\) & 0.87 & 0.87 & 0.91 & 0.87 \\  & \(p(\chi 2)\) CA vs. & – & 0.587 & \(5.49\times\) 10\({}^{-14}\) * & 0.768 \\  & S/A vs. & – & – & 6.42 \(\times\) 10\({}^{-15}\) * & 0.631 \\ \cline{2-5}  & \multicolumn{1}{c}{**Parameter**} & \multicolumn{4}{c}{**The Independent Action (IA)-Based Module**} \\  & & **IA** & **S/A** & **DR** & **DL** \\ \cline{2-5}
1 h & \(a\) & – & –5.325 & –3.654 & –15.34 \\  & \(b\) & – & – & –5.365 & –8.324 \\  & \(R^{2}\) & 0.35 & 0.78 & 0.85 & 0.99 \\  & \(p(\chi 2)\) IA vs. & – & 1.5 \(\times\) 10\({}^{-282}\) * & 0.000 * & 0.000 * \\  & S/A vs. & – & – & 1.72 \(\times\) 10\({}^{-5}\) * & 1.52 \(\times\) 10\({}^{-13}\) * \\
24 h & \(a\) & – & –1.478 & –7.544 & –1.812 \\  & \(b\) & – & – & 10.366 & 0.312 \\  & \(R^{2}\) & 0.86 & 0.97 & 0.92 & 0.87 \\  & \(p(\chi 2)\) IA vs. & – & 1.1 \(\times\) 10\({}^{-07}\) * & 3.89 \(\times\) 10\({}^{-23}\) * & 6.48 \(\times\) 10\({}^{-7}\) * \\  & S/A vs. & – & – & 4.66 \(\times\) 10\({}^{-16}\) * & 0.574 \\ \hline \hline \end{tabular}  
* 1 h: 1 h exposure period; 24 h: 24 h exposure period; CA: the concentration addition model; IA: the independent addition model; a and b: interaction parameters; S/A: the model describing synergy or antagonism with respect to the reference model has one interaction parameter (\(a\)); DR: the model describing dose ratio-dependent deviations from the reference model has two interaction parameters (\(a\) and \(b\)); DR: the model describing dose ratio-dependent deviations from the reference model has two interaction parameters (\(a\) and \(b\)); DR: the coefficient of determination; \(p(\chi 2)\): the outcome of the likelihood ratio test; vs.: versus, which was used to show the comparison between two models; \(\sim\): not applicable. *: Significant at the 5\(\%\) significance level.  
\end{table} 
Table 3: Fitting results of the toxicity data of the bimetallic mixture (Cd + ZnO) to the four models describing deviations from concentration addition (CA) and independent action (IA) of _Euplotes aeidiculatus_ using the MixTOX tool. variance in the data. A negative \"_a_\" parameter indicates synergism, while a positive \"_p_\" parameter indicates antagonism at specific dose ratios. When the data were examined, strong antagonism was observed when Cd toxicity was particularly high. These results are consistent with the results of the IA sub-modules (Table 3). Antioxidant Properties of E. aediculatus Extracts Treated with Single and Binary Mixture at 1 h and 24 h  
The total phenolic content has been identified as the major naturally occurring antioxidant component in the extracts of the freshwater ciliate _E. aediculatus_. In this study, the total phenolic content (TPC) of ciliate extracts treated with single metals (Cd, Zn, and ZnO) and bimetallic mixtures (Cd + Zn and Cd + ZnO) at 1 h and 24 h was evaluated. The Folin-Ciocalteu Method was used for the measurement, revealing concentrations ranging from 95.44 to 148.65 \(\upmu\)g/L. _E. aediculatus_ showed the lowest phenolic value at concentrations of 1 TU ZnO, while the maximum was observed at 0.5 TU Cd and 0.5 TU Zn at 1 h and 24 h, respectively, with a significant level of \(p\) < 0.001. In the case of single-metal treatments, the TPC of _E. aediculatus_ was higher after 24 h of exposure compared to 1 h. In the bimetallic mixture treatments, the TPC was lower in the binary mixture of Cd + ZnO compared to the binary mixture of Cd + Zn at 24 h (Figure 1).