Single-substance and mixture toxicity of five pharmaceuticals and personal care products to marine periphyton communities.

The single-substance and mixture toxicity of five pharmaceuticals and personal care products (fluoxetine, propranolol, triclosan, zinc-pyrithione, and clotrimazole) to marine microalgal communities (periphyton) was investigated. All compounds proved to be toxic, with median effective concentration values (EC50s) between 1,800 nmol/L (triclosan) and 7.2 nmol/L (Zn-pyrithione). With an EC50 of 356 nmol/L, the toxicity of the mixture falls into this span, indicating the absence of strong synergisms or antagonisms. In fact, a comparison with mixture toxicity predictions by the classical mixture concepts of concentration addition and independent action showed a good predictability in the upper effect range. However, the mixture provoked stimulating effects (hormesis) in the lower effect range, hampering the application of either concept. An independent repetition of the mixture experiment resulted in a principally similar concentration-response curve, again with clear hormesis effects in the lower range of test concentrations. However, the curve was shifted toward higher effect concentrations (EC50 1,070 nmol/L), which likely is due to changes in the initial species composition. Clear mixture effects were observed even when all five components were present only at their individual no-observed-effect concentrations (NOECs). These results show that, even with respect to mixtures of chemically and functionally dissimilar compounds, such as the five pharmaceuticals and personal care products investigated, environmental quality standards must take possible mixture effects from low-effect concentrations of individual compounds into consideration.

Mixture toxicity from photosystem II inhibitors on microalgal community succession is predictable by concentration addition.

The typical pollution situation involves chemical mixtures, and assessing the risks of single chemicals one at a time is not sufficient. Concentration addition (CA) has been suggested as a predictive tool in mixture ecotoxicology. The accuracy of CA for mixtures of similarly acting chemicals has been demonstrated under relatively simple biological conditions in single-species tests. To consider the high diversity of interconnected species in ecosystems, one must evaluate CA on a community level of biological organization. We sampled marine periphyton communities from the west coast of Sweden and exposed them to photosystem II (PSII) inhibiting herbicides for 4 d in the SWIFT test, a semistatic, small-scale laboratory test. During this time, the communities went through an ecological succession, influenced by the toxicants in a concentration-dependent manner. Multidimensional scaling was used to assess similarities in the effects of two different sets of PSII inhibitors on pigment profiles, which reflects the taxonomic structure and the physiological status of the microalgal community. One mixture of structurally congeneric phenylureas and one mixture of non-congeneric PSII inhibitors were tested. All PSII inhibitors and their mixtures caused similar changes in the pigment profiles, demonstrating that they not only have a similar biochemical mechanism of action but also are similarly acting on a community level. Concentration addition accurately predicted the effects of both mixtures over the entire effect range. This demonstrates that chemical congenericity is not required for a high predictive power of CA. Instead, in perfect analogy to the situation in single-species tests, a similar mode of action is a sufficient prerequisite for a successful application of CA.

Toxicity of the pharmaceutical clotrimazole to marine microalgal communities.

Clotrimazole belongs to the group of 14alpha-demethylase inhibiting fungicides. It is widely used in human and veterinary medicine and has been identified as a priority pollutant for the marine environment. However, the toxicity of clotrimazole to marine primary producers is largely unknown. We therefore sampled natural microalgal communities (periphyton) and exposed them to concentration series of clotrimazole over 4 days. 50 pmol/L clotrimazole caused a concentration-dependent accumulation of C14alpha-methylated sterol precursors, which coincided with a decrease in algal-specific C14-desmethyl sterols. This indicates an inhibition of algal 14 alpha-demethylases already at environmental concentrations. A clotrimazole concentration of 500 pmol/L reduced total sterol content to 64% of control level. Community chlorophyll a content was affected by clotrimazole in a bi-phasic manner with first reductions becoming visible at 500 pmol/L, along with indications of an altered cycling of photoprotective xanthophyll pigments. Concentrations of 10-100 nmol/L and higher caused large reductions in community growth, and changed community pigment profiles in a concentration-dependent monotonous manner. The study further indicated that diatoms use obtusifoliol as a natural substrate for 14alpha-demethylase, just as higher plants do but also utilize norlanosterol.