In the environment, microalgae are exposed to a multitude of stressors simultaneously, inducing physiological adjustments. It is well documented that both phosphorus (P) limitation and trace metals exposure affect microalgal physiology. However, investigations regarding the combination of both P limitation and excess trace metals still deserve attention. In the present study, we evaluated the changes in photosynthetic parameters in the green microalga Ankistrodesmus densus acclimated to different P concentrations prior to exposure to Cd. Our results indicate that different concentrations of P in the medium were responsible for significant changes in some parameters, especially those related to photoprotection mechanisms. Cadmium also altered some of these variables in all P scenarios, and greater damage (i.e., synergism) was observed in the combination P-limited and high Cd, with all the evaluated parameters affected under the adverse scenario. Among the parameters analyzed, rapid light curves were the most sensitive to exposure of one or the combination of both stressors (Cd and P limitation). Based on our data, we suggest that P-limited algae activated photoprotective mechanisms as a response to nutrient limitation, especially at the most limited condition. The addition of Cd did not change linearly the parameters related to photoprotection mechanisms under P-limitation, i.e., synergism was observed in the intermediate P-limitation combined with Cd, while in the most P-limited, P seems to be the driving force affecting these mechanisms. Based on our results, we suggest the use of rapid light curves as a tool to complement the assessment of the impacts of stressors, such as metals, in ecotoxicological studies.

Plant-derived materials as environmentally friendly biosorbents to remove heavy metals from wastewater have been extensively studied. However, the chemical oxygen demand (COD) increase caused by the plant-derived biosorbent has not been considered previously. In this study, water hyacinth was used as biosorbent to remove Cd(II) from wastewater. About 66% of Cd(II) was removed by the biosorbent with a maximum biosorption capacity (qₘₐₓ) of 21.6 mg g⁻¹. However, the COD of the filtrate increased from 0 to 292 mg L⁻¹ during this process. Subsequently, endophytes, microalgae and the microalgae-endophyte symbiotic system (MESS) were assessed for the simultaneous Cd(II) and COD removal. Among these three systems, the MESS achieved the best performance. After 3 d of inoculation, the extent of total Cd(II) removal increased to 99.2% while COD decreased to 77 mg L⁻¹. This study provides a new insight into the application of a plant-derived biosorbent in combination with microalgae and endophytes for the effective treatment of heavy metal-bearing wastewater.

The increasing water pollution caused by the presence of nano- and microplastics has shown a need to pursue solutions to remediate this problem. In this work, an extracellular polymeric substance (EPS) producing freshwater Cyanothece sp. strain was exposed to nano- and microplastics. The bioflocculant capacity of the biopolymer produced was evaluated. The influence of different concentrations (1 and 10 mg L⁻¹) of polystyrene nano- and microplastics in the extracellular carbohydrates and in the EPS production was studied. The presence of nano- and microplastics induced a negative effect on the microalgal growth (of up to 47%). The results show that the EPS produced by Cyanothece sp. exhibits high bioflocculant activity in low concentrations. Also, the EPS displayed very favourable characteristics for aggregation, as the aggregates were confirmed to consist of microalga, EPS and both the nano- and microplastics. These results highlight the potential of the microalgal-based biopolymers to replace hazardous synthetic flocculants used in wastewater treatment, while aggregating and flocculating nano- and microplastics, demonstrating to be a multi-purposed, compelling, biocompatible solution to nano- and microplastic pollution.

The effects of increasing concentrations of suspended mineral coal dust on the energetic physiology of the Caribbean scallop Argopecten nucleus were studied, at a concentration range that is environmentally relevant and representative of areas proximate to coal loading and shipping ports. Adult hatchery-produced animals were exposed to different concentrations of coal dust, i.e. 0, 2, 9 and 40 mg L⁻¹. At increasing concentrations of coal dust, the rates of filtration and pseudofeces production increased, while the rates of ingestion and absorption remained constant. The rates of oxygen consumption and ammonium excretion decreased, as well as the absorption efficiency and the scope for growth. Suspended coal dust particles, at concentrations higher than or equal to 2 mg L⁻¹, were ingested preferentially over microalgae by A. nucleus, causing reductions in its absorption capability, metabolism and in the amount of energy for growth and reproduction, thus generating physiological stress.

Australian tropical freshwaters can experience extreme seasonal variability in rainfall and run off, particularly due to pulse events such as storms and cyclones. This study investigated how seasonal variability in dissolved organic matter (DOM) quality impacted the chronic toxicity of copper to a tropical green alga (Chlorella sp.) in the presence of two concentrations of DOM (low: ∼2 mg C/L; high: ∼10 mg C/L) collected from three tropical waters. Copper speciation and lability were explored using diffusive gradients in thin-films (DGT) and modelled maximum dynamic concentrations (cᵈʸⁿₘₐₓ) using data derived from the Windermere Humic Aqueous Model (WHAM VII). Relationships between copper lability and copper toxicity were assessed as potential tools for predicting toxicity. Copper toxicity varied significantly with DOM concentration, source and season. Copper toxicity decreased with increasing concentrations of DOM, with 50% growth inhibition effect concentrations (EC₅₀) increasing from 1.9 μg Cu/L in synthetic test waters with no added DOM (0.34 mg C/L) up to 63 μg Cu/L at DOM concentrations of 9.9 mg C/L. Copper toxicity varied by up to 2-fold between the three DOM sources and EC₅₀ values were generally lower in the presence of wet season DOM compared to dry season DOM. Linear relationships between DGT-labile copper and dissolved copper were significantly different between DOM source, but not concentration or season. Modelled cᵈʸⁿₘₐₓ consistently under-predicted labile copper in high DOM treatments compared to DGT measurements but performed better in low DOM treatments, indicating that this method is DOM-concentration dependent. Neither speciation method was a good surrogate for copper toxicity in the presence of different sources of natural DOM. Our findings show that DOM source and season, not just DOM concentration, affect copper toxicity to freshwater biota. Therefore, DOM quality should be considered as a toxicity-modifying factor for future derivation of bioavailability-based site-specific water quality guideline values.

In recent years, antibiotic pollution has become worse, especially in China. In this study, the ecotoxicological effects of four frequently used antibiotics with different lipophilic degrees (log Kow) (sulfadiazine (SD), sulfamethazine (SM2), enrofloxacin (ENR), and norfloxacin (NOR)) at four concentrations of 1, 5, 20, and 50 mg L⁻¹ were examined using batch cultures of green alga Chlorella vulgaris and cyanobacterium Chrysosporum ovalisporum for 16 days based on changes in chlorophyll fluorescence parameters (chl a, Fv/Fm, and ΦPSII) and responses of the antioxidant system. Besides, the antibiotics removal efficiencies of the two microalgae were investigated. Sulfonamides (SD and SM2) had no significant inhibitory effect on the growth of C. ovalisporum, but had an inhibitory effect on C. vulgaris, whereas fluoroquinolones (ENR and NOR) significantly inhibited C. ovalisporum. The activities of superoxide dismutase, catalase, and glutathione reductase suggested that C. vulgaris was more tolerant to these antibiotics than C. ovalisporum. The increased malondialdehyde level in both algae indicated their tolerance against antibiotics. When compared with C. ovalisporum, C. vulgaris presented better capacity to remove antibiotics. In summary, the four antibiotics exerted time- or concentration-dependent ecotoxicological effects on the microalgae examined, whereas the microalgae could remove the antibiotics based on the log Kow of the antibiotics. The findings of this study contribute to effective understanding of the ecotoxicological effects of antibiotics and their removal by microalgae.

Copper oxide nanoparticles (CuO NP) have been produced on a large scale due to their economically interesting thermophysical properties. This heightens the concern about risks they may pose on their release into the environment, possibly affecting non-target organisms. Microalga are important organisms in ecotoxicological studies as they are at the base of the aquatic food chain, but information about their biochemical and photosynthetic changes in response CuO NP are still scarce. We studied the effects of CuO NP in Raphidocelis subcapitata using morphological, photosynthetic and biochemical biomarkers. Our results showed that the NP affected microalgal population growth with 0.70 mg Cu L⁻¹ IC₅₀–₉₆ ₕ (inhibition concentration). Based on predicted environmental concentrations of Cu NPs in aquatic environments, our results indicate potential risks of the NP to microalgae. Algal cell size, granularity and photosynthetic efficiencies were affected by the CuO NP at 0.97 and 11.74 mg Cu L⁻¹. Furthermore, lipid metabolism was affected mostly at the highest NP concentration, but at environmentally relevant values (0.012 and 0.065 mg Cu L⁻¹) the production of sterols (structural lipids) and triacylglycerols (reserve lipid) increased. Moreover, we found evidence of cell membrane impairment at the highest CuO NP concentration, and, as a photosynthetic response, the oxygen evolving complex was its main site of action. To the best of our knowledge, this is the first study to date to investigate microalgal lipid composition during CuO NP exposure, showing that it is a sensitive diagnostic tool. This research demonstrated that CuO NP may affect the physiology of R. subcapitata, and because they were observed in a primary producer, we foresee consequences to higher trophic levels in aquatic communities.

Microcystis aeruginosa is one of the main species of cyanobacteria that causes water blooms. M. aeruginosa can release into the water several types of microcystins (MCs), which are harmful to aquatic organisms and even humans. However, few studies have investigated the hepatotoxicity of M. aeruginosa itself in zebrafish in environments that simulate natural aquatic systems. The objective of this study was to evaluate the hepatotoxicity of M. aeruginosa in adult zebrafish (Danio rerio) after short-term (96 h) exposure and to elucidate the potential underlying mechanisms. Distinct histological changes in the liver, such as enlargement of the peripheral nuclei and sinusoids and the appearance of fibroblasts, were observed in zebrafish grown in M. aeruginosa culture. In addition, antioxidant enzyme activity was activated and protein phosphatase (PP) activity was significantly decreased with increasing microalgal density. A proteomic analysis revealed alterations in a number of protein pathways, including ribosome translation, immune response, energy metabolism and oxidative phosphorylation pathways. Western blot and real-time PCR analyses confirmed the results of the proteomic analysis. All results indicated that M. aeruginosa could disrupt hepatic functions in adult zebrafish, thus highlighting the necessity of ecotoxicity assessments for M. aeruginosa at environmentally relevant densities.

The widespread application of commercial TiO₂ NPs inevitably leads to their release into environmental waters through various ways. TiO₂ NPs released into water might be absorbed by and react with periphytic biofilms, which are a kind of aquatic environmental media of important ecological significance, and influence the physiological activity and ecological function of periphytic biofilms. This study investigated the effects of exposure to 1 mg/L and 5 mg/L of TiO₂ NPs on periphytic biofilms cultured indoors. After a 10-day exposure to TiO₂ NPs, the growth (measured by chlorophyll-a content) of microalgal community was inhibited greatly (more than 60%); however, the primary production (indicated by quantum yield) of periphytic biofilms maintained changeless. As for bacteria, TiO₂ NP-exposure increased the bacterial diversity and altered the composition structure. Significant changes were observed in the bacterial communities at the class level, mainly including Alphaproteobacteria, Gammaproteobacteria, Cytophagia, Flavobacteriia, Sphingobacteriia, Synechococcophycideae and Oscillatoriophycideae. The enhancement of metabolic activities (the production of extracellular polymeric substances, especially proteins content increased by 48.51%) of periphytic biofilms was a resistance mechanism to toxicity of NPs. As for extracellular enzyme activities of periphytic biofilms, alkaline phosphatase activity was inhibited (22.43%) after exposed to 5 mg/L of TiO₂ NPs, which posed a threat to phosphorus metabolism of periphytic biofilms. Overall, this study demonstrated that 1 mg/L and 5 mg/L of TiO₂ NPs negatively influenced physiological activities and ecological functions of periphytic biofilms, highlighting that the ecological risks of TiO₂ NPs should be paid attention to.

Microalgae are commonly used in ecotoxicity testing due to their ease of culturing and rapid cell division rates. These tests generally utilise a single species of algae; however, microalgae occur in the environment as complex communities of multiple species. To date, routine multispecies toxicity tests using tropical microalgae have not been available. This study investigated four tropical freshwater microalgal species for use in a chronic multispecies toxicity test based on the population growth (cell division) rate: Pediastrum duplex, Monoraphidium arcuatum, Nannochloropsis-like sp. and Chlorella sp. 12. Flow cytometric analysis identified the different fluorescence and light scattering properties of each algal species and quantified each species within multispecies mixtures. Following optimisation of test media nutrients and pH, a toxicity testing protocol was developed with P. duplex, M. arcuatum and Nannochloropsis-like sp. There were no significant differences in growth rates of each alga when tested over 72 h as single species or in multispecies mixtures. Atrazine and imazapic, two herbicides with different modes of action, were used to assess the sensitivity of the multispecies toxicity test. Atrazine was toxic to all species with 72-h IC10 values of 7.2, 63 and 280 μg/L for P. duplex, M. arcuatum and Nannochloropsis-like sp. respectively, while imazapic was not toxic to any species at concentrations up to 1100 μg/L. The toxicity of atrazine and imazapic to each microalgal species in the multispecies toxicity test was the same as that determined from single-species toxicity tests indicating that the presence of these microalgae in a mixture did not affect the toxicity of these two herbicides. This study is the first to develop a multispecies tropical microalgal toxicity test for application in freshwaters. This time- and cost-effective tool can be utilised to generate data to assist environmental decision making and to undertake risk assessments of contaminants in tropical freshwater environments.