Transcriptional response of a green alga (Raphidocelis subcapitata) exposed to triclosan: photosynthetic systems and DNA repair.

Recent studies show that triclosan (TCS) exposure causes reduction in pigments, suppression of photosynthesis, and induction of oxidative stress at the physiological level, resulting in morphological alteration and growth inhibition in algae including Raphidocelis subcapitata (R. subcapitata, a freshwater model green alga). However, the underlying molecular mechanisms remain to be elucidated, especially at environmentally relevant concentrations. The present study uncovered the transcriptional profiles and molecular mechanisms of TCS toxicity in R. subcapitata using next-generation sequencing. The algal growth was drastically inhibited following a 7-day exposure at both 75 and 100 µg/L TCS, but not at 5 µg/L (environmentally realistic level). The transcriptomic analysis shows that molecular signaling pathways including porphyrin and chlorophyll metabolism, photosynthesis - antenna proteins, and photosynthesis were suppressed in all three TCS treatments, and the perturbations of these signaling pathways were exacerbated with increased TCS exposure concentrations. Additionally, signaling of replication-coupled DNA repair was only activated in 100 µg/L TCS treatment. These results indicate that photosynthesis systems were sensitive targets of TCS toxicity in R. subcapitata, which is distinct from the inhibition of lipid synthesis by TCS in bacteria. This study provides novel knowledge on molecular mechanisms of TCS toxicity in R. subcapitata.

Involvement of oxidative stress in the sensitivity of two algal species exposed to roxithromycin.

Algal species Raphidocelis subcapitata and Chlorella vulgaris are commonly used to test the chemicals with an antibacterial mode of action during marketing authorization process. However, significant differences in the sensitivity toward antibiotic exposure have been reported. The selection of an inappropriate test species would thus underestimate the environmental hazard of target chemicals and pose a potential threat to the ecosystem. Since oxidative stress is a crucial factor determining the inhibition of algal growth, an investigation on oxidative stress and antioxidant defense mechanisms in these two species was performed to explore its roles in species sensitivity. Here, roxithromycin (ROX), a macrolide antibiotic extensively used to treat respiratory, urinary and soft tissue infections, was used for testing. After 7 days exposure to ROX at the low (0.01 mg L-1) and high (0.09 mg L-1) concentrations, R. subcapitata was inhibited while the growth of C. vulgaris was stimulated. We investigated the roles of oxidative stress in algae by measuring the oxidative stress biomarkers (MDA), non-enzymatic antioxidants (GSH), and antioxidant enzymes (SOD, CAT, GP, GST). The results suggested that when the growth of algae is inhibited, MDA content as well as activities of oxidative stress enzymes would increase, and thus, activating the antioxidant system. On the contrary, it was inferred that when the growth is stimulated, MDA content and oxidative stress enzymes activities would decrease.

Transcriptomic analysis and transgenerational effects of ZnO nanoparticles on Daphnia magna: Endocrine-disrupting potential and energy metabolism.

The widespread application of zinc oxide nanoparticles (ZnO NPs) has raised concerns over the adverse effects on aquatic species. In this study, transcriptomic analysis was applied to evaluate the chronic toxicity of ZnO NPs on the freshwater invertebrate Daphnia magna and the intergenerational effects were then further investigated. Parent daphnia (F0) were exposed to ZnO NPs at 3, 60, and 300 µg L-1 for 21 days. ZnO NPs significantly inhibited the reproduction (first pregnancy and spawning time, total number of offspring) and growth (molting frequency and body length) of F0. Here, differentially expressed genes (DEGs) involved in lysosomal and phagosome, energy metabolism and endocrine disruption pathways were significantly downregulated. Furthermore, disruption on the transport and catabolic processes probably resulted in the particle accumulation. The inhibited pathways related to energy metabolism may partially account for the body length, molting and reproductive restriction. The suppression of growth and reproduction may attribute to the down-regulation of insulin secretion and ovarian steroidogenesis pathways, respectively. Partial recovery of growth and reproductive inhibition in F1 - F3 descended from the F0 generation exposure did not support constant transgenerational effects. This study unravels the molecular mechanisms and transgenerational consequences of the toxicity of nanoparticles on Daphnia.

Antibiotic Toxicity Isolated and as Binary Mixture to Freshwater Algae Raphidocelis subcapitata: Growth Inhibition, Prediction Model, and Environmental Risk Assessment.

Antibiotics in aqueous environments can have extremely adverse effects on non-targeted organisms. However, many research projects have only focused on the toxicological evaluation of individual antibiotics in various environments. In the present work, individual and binary mixture toxicity experiments have been conducted with the model organism Raphidocelis subcapitata (R. subcapitata), and a mixture concentration-response curve was established and contrasted with the estimated effects on the basis of both the concentration addition (CA) and the independent action (IA) models. In addition, different risk assessment methods were used and compared to evaluate the environmental risk of binary mixtures. The toxic ranking of the selected antibiotics to R. subcapitata was erythromycin (ERY) > sulfamethoxazole (SMX) > sulfamethazine (SMZ). In general, the conclusion of this study is that the adverse effects of binary mixtures are higher than the individual antibiotics. The CA model and RQSTU are more suitable for toxicity prediction and risk assessment of binary mixtures. This study reveals the potential ecological risks that antibiotics and their mixtures may pose to water ecosystems, thus providing scientific information for environmental quality regulation.

Prediction of adverse effects of effluents containing phenolic compounds in the Ba River on the ovary of fish (Hemiculter leucisculus) using transcriptomic and metabolomic analyses.

The aim of this work was to evaluate the endocrine disrupting effects on the ovarian development of sharpbelly (Hemiculter leucisculus) caused by effluents containing phenolic compounds. This was achieved using integrated transcriptomic and metabolomic analyses, along with histopathological examinations. Sharpbelly, an indigenous freshwater fish widely distributed in East Asia, were collected by pole fishing from three sampling sites in the Ba River. These sampling sites include a mid-stream site near a wastewater outfall and a reference site located upstream and a far field comparison site located downstream. In sharpbelly collected near the wastewater discharge, the oocyte development was activated, compared to the other two sites. Histopathological alterations in the fish ovaries were likely due to the upregulated steroid hormone biosynthesis process, as suggested by the differentially expressed genes (e.g., hsd3b, hsd17b1) and differentially accumulated metabolites (e.g., pregnenolone). Additionally, under the stress of effluents containing phenolic compounds, genes related to the signaling pathways for oxidative phosphorylation and leukocyte transendothelial migration were dysregulated, suggesting the potential induction of inflammation and several ovarian diseases. Overall, these findings suggest that effluents containing phenolic compounds influence ovary development and reproductive function of female sharpbelly. Whether there is any resulting dysfunction of folliculogenesis, abnormality of ovulation, production of premature eggs and/or potential induction of ovarian cancers remains to be determined by further studies, for a better evaluation on effluents containing phenolic compounds to the fish fertility and the health of their offspring, and even the stability of the wild fish population. Notably, the integration of transcriptomics and metabolomics can complement the routine chemical analysis to comprehensively monitor the effects of wastewater treatment plant effluents on the health of wild fish.