Disentangling the Impacts of PAHs, Microplastics, and Sediment Resuspension on Algal Physiology: A Partial Least Squares Structural Equation Modeling Approach

Environmental stressors, such as contaminants and physical factors, rarely act in isolation, and studying their joint effects provides a more accurate reflection of real-world scenarios. To capture these interactions and disentangle the direct and indirect influences on algal responses, we applied partial least squares structural equation modeling (PLS-SEM), allowing us to reveal the hierarchical relationships among stressors and their cumulative impact on algal physiology. We examined combined effects of microplastics (MP; presence/absence), polycyclic aromatic hydrocarbons (PAHs; a mixture of acenaphthene, fluorene, phenanthrene, and fluoranthene at a total chemical activity in the sediment of 0 or 0.14), and sediment resuspension (turbidity: 0.8-3.9 NTU) on <i>Ceramium tenuicorne</i>, a coastal macroalga that is likely to encounter all these stressors in its natural habitats. Mechanical mixing at two intensities (low and high) was applied as an experimental treatment to induce resuspension. The analysis separated the effects of mechanical mixing and turbidity, given their nonlinear relationship, as stronger mechanical mixing did not consistently result in proportional turbidity increases. The algal physiological responses were evaluated using changes in pigment composition (Chl <i>a</i>, Chl <i>c</i>, and carotenoids), photosystem II (PSII) performance, total antioxidant capacity, and algal stoichiometry measured as elemental (%C, %N, %H, and C/N) ratios. We found that PAH exposure was the main suppressor of pigment concentrations and PSII performance, underscoring the mechanisms of its adverse effects on the photosynthetic machinery and nutrient assimilation. Moreover, stronger turbulence further decreased pigment concentrations, while sediment resuspension increased antioxidant capacity in algae, possibly due to physical damage from abrasion and scouring. We also found that MP addition significantly increased turbidity, thus aggravating the effects of the sediment resuspension. In conclusion, we provide a mechanistic explanation of how the combined exposure to MPs, PAHs, and sediment resuspension can impact pigment composition, photosynthesis, and stoichiometry of the algae, leading to decreased productivity.