In developing countries, wastewater treatment is confined to secondary systems. Hence even after treatment, wastewater effluent has a high level of nutrients which causes eutrophication and has destructive impacts on receiving bodies. Literature reveals that phycoremediation can be the best solution to address the problem faced but is time-consuming, ranging from days to weeks. Hence, the present study aimed to determine an optimum detention time for the microalgal system to treat domestic wastewater. The retention time for treatment in the study was divided into an aeration and settling periods. During the study, aeration time varied from 2 hours to 24 hours, followed by 1-hour settling period for each aeration time. Optimum detention time for microalgal treatment was obtained at 11 hours of detention time (10 hours aeration and 1-hour settling). Parameters analyzed during the study were pH, EC, TS, TSS, TDS, nitrate, phosphate, ammonia, COD and DO. However, the main focus was on nutrients (phosphate and ammonia) and organics (COD) removal while determining the optimum detention time. Maximum removal efficiency obtained for COD, ammonia and phosphate for non-filtered effluent was 75.61%, 90.63% and 83.29%, respectively. However, removal efficiency further increased for filtered effluents to 86.34%, 100% and 91.12% for COD, ammonia and phosphate, respectively. Algal treatment offers an ecologically safe and more affordable system for nutrient removal and eliminates the need for tertiary treatment.

This study presents a novel algal-based toxicity test suitable for simple and rapid assessment of heavy metal (Hg2+, Cr6+, Cd2+, Pb2+, or As3+)-contaminated water. A closed-system kit-type algal assay was developed using Chlorella vulgaris. Toxicity was assessed by oxygen evolution in the gaseous phase of the assay kits, which was measured via a needle-type oxygen sensor. Initial cell density, light intensity, and exposure time that enabled favorable test performance for the algal assay kits were 103 cells/mL, 250 μmol m-2s-1, and 18 h, respectively. Results from the heavy metal toxicity tests demonstrate that Hg2+, Cr6+, Cd2+, and Pb2+ are more toxic in inhibiting algal photosynthetic activity than As3+. The 18 h half-maximum effective concentrations (EC50) for Hg2+, Cr6+, Cd2+, Pb2+, and As3+ were determined to be 31.3 ± 0.5, 179.6 ± 7.5, 301.3 ± 6.1, 476.1 ± 10.5, and 2184.1 ± 31.1 μg/L, respectively. A strong correlation between oxygen concentrations in the headspace of the assay kits and chlorophyll a production indicates that oxygen evolution in the gaseous phase is able to represent algal photosynthetic activity and serve as the end-point in algal toxicity tests. High test sensitivity and reproducibility as well as an easy test protocol and rapid processing time make the algal assay kit a suitable tool for simple and rapid toxicity testing of heavy metal-contaminated water.

To compare aquatic organisms’ responses to the toxicity of copper oxide (CuO) nanoparticles (NPs) with those of CuO microparticles (MPs) and copper (Cu) ions, a global metabolomics approach was employed to investigate the changes of both polar and nonpolar metabolites in microalga Chlorella vulgaris after 5-day exposure to CuO NPs and MPs (1 and 10 mg/L), as well as the corresponding dissolved Cu ions (0.08 and 0.8 mg/L). Unchanged growth, slight reactive oxygen species production, and significant membrane damage (at 10 mg/L CuO particles) in C. vulgaris were demonstrated. A total of 75 differentiated metabolites were identified. Most metabolic pathways perturbed after CuO NPs exposure were shared by those after CuO MPs and Cu ions exposure, including accumulation of chlorophyll intermediates (max. 2.4–5.2 fold), membrane lipids remodeling for membrane protection (decrease of phosphatidylethanolamines (min. 0.6 fold) and phosphatidylcholines (min. 0.2–0.7 fold), as well as increase of phosphatidic acids (max. 1.5–2.9 fold), phosphatidylglycerols (max. 2.2–2.3 fold), monogalactosyldiacylglycerols (max. 1.2–1.4 fold), digalactosylmonoacylglycerols (max. 1.9–3.8 fold), diacylglycerols (max. 1.4 fold), lysophospholipids (max. 1.8–3.0 fold), and fatty acids (max. 3.0–6.2 fold)), perturbation of glutathione metabolism induced by oxidative stress, and accumulation of osmoregulants (max. 1.3–2.6 fold) to counteract osmotic stress. The only difference between metabolic responses to particles and those to ions was the accumulation of fatty acids oxidation products: particles caused higher fold changes (particles/ions ratio 1.9–3.0) at 1 mg/L and lower fold changes (particles/ions ratio 0.4–0.7) at 10 mg/L compared with ions. Compared with microparticles, there was no nanoparticle-specific pathway perturbed. These results confirm the predominant role of dissolved Cu ions on the toxicity of CuO NPs and MPs, and also reveal particle-specific toxicity from a metabolomics perspective.

As an essential micronutrient for animals with a narrow range between essentiality and toxicity, selenium (Se) usually coexists with chromium (Cr) in contaminated aquatic environments. This study investigated effects of three diets (Microcystis aeruginosa, Chlorella vulgaris and biofilms) exposed to Se or/and Cr on Aedes albopictus as a vector for the aquatic-terrestrial transfer of Se and Cr. Se(IV)-exposed mosquitoes concentrated Se up to 66-fold faster than Se(VI)-exposed ones, corresponding to the greater Se enrichment in Se(IV)-treated diets. Analysis using synchrotron-based X-ray absorption spectroscopy (XAS) showed that Se(0) (61.9–74.6%) dominated Se(VI)-exposed mosquitoes except for the C. vulgaris-fed larvae (organo-Se, 94.0%), while organo-Se accounted for 93.3–100.0% in Se(IV)-exposed mosquitoes. Cr accumulation in larvae (56.40–87.24 μg Cr/g DW) or adults (19.41–50.77 μg Cr/g DW) was not significantly different among all Cr(VI) treatments, despite varying diet Cr levels. With Cr(0) being dominant (57.7–94.0%), Cr(VI)-exposed mosquitoes posed little threat to predators. Although mosquitoes exposed to Se or Cr had shorter wings, adults supplied with C. vulgaris or biofilms co-exposed to Se(VI) and Cr(VI) had wings significantly (1.1–1.2 fold) longer than Se(VI) only exposed ones. Overall, our study reveals the role of Ae. albopictus in transferring waterborne Se and Cr from the contaminated aquatic ecosystem to the terrestrial ecosystem with the resulting eco-risks to wildlife in both ecosystems.

In the present study, we evaluate our previously developed non-destructive soil algal toxicity method using species from a different class of algae; Class Trebouxiophyceae (Chlorella vulgaris and Chlorella sorokiniana), and directly measure the photosynthetic activity of these species adsorbed onto the soil as a new toxicity endpoint. This study shows that non-destructive soil algal toxicity method is applicable to non-specific test species, including those of Class Trebouxiophyceae as well as Class Chlorophyceae (Chlorococcum infusionum and Chlamydomonas reinhardtii). Furthermore, by performing photosynthesis image analysis, we verify that it is possible to measure the photosynthetic activity of soil algae Chlorella vulgaris adsorbed onto soils without the need to extract algal cells from the soil. We propose that the non-destructive soil algal toxicity method represents a novel technique for 1) evaluating pollutants in soil using non-specific algae and 2) conveniently and rapidly assessing the photosynthetic activity of soil algae Chlorella vulgaris adsorbed onto soil as a new toxicity endpoint.

Boscalid is one of the most frequently detected pesticides in main coastal estuaries in California, with concentrations as high as 36 μg/L. However, ecotoxicology information about boscalid to aquatic organisms is scarce. To investigate toxic effects and mechanisms of boscalid on freshwater algae Chlorella vulgaris (C. vulgaris), C. vulgaris were exposed to a range of boscalid concentrations (0, 0.8, 1.6, 2.4 and 3.2 mg/L) for 96 h to study the changes in photosynthetic pigment contents, responses of the antioxidant enzyme system and alterations in endogenous substances. Results indicated that the growth of algae and the content of chlorophyll and carotenoids were significantly inhibited by 1.6 mg/L boscalid. Reactive oxygen species (ROS) and oxidative damage of C. vulgaris could be induced by boscalid, in accordance with significant changes in ROS levels and a series of antioxidant enzyme activities. Moreover, the alterations in endogenous substances showed that boscalid could affect photosynthesis and energy metabolism of C. vulgaris. These results demonstrated that boscalid could induce impacts on C. vulgaris mainly through disturbing the photosynthesis, oxidative damage and energy metabolism. The present study provided a better understanding of the negative effects and mechanisms of bosaclid in microalgae.

Androstenedione (ADSD) was the main androgen detected in wastewaters. Chlorella was the most widely used plant in biological wastewater treatment process. In order to understand the toxicological response of chlorella to ADSD contamination, we used the weighted gene co-expression network analysis (WGCNA) method to systematically analyze the gene regulatory networks of chlorella after ADSD treatments. Total of 25 modules was identified from gene co-expression networks, and the turquoise module were selected for GO and KEGG enrichment analysis. Results showed that most hub genes were associated with chloroplast organizations or photosystems processes. Among them, the expressions profiles of hcar, nol, pao and sgr genes were highly correlated to the content fluctuations of chlorophylls after different ADSD treatments. All these results demonstrated that chlorophylls play a key role in preventing cell damage of chlorella caused by ADSD contamination. Besides, we proposed a possible chlorophyll metabolism pathway in chlorella response to ADSD contamination.

The effects of algae on the removal of contaminant by iron sulfide (FeS) are still unknown. Chlorella vulgaris (CV), a remarkable algal specie, was used to prepare the CV-supported FeS (CV-FeS) and to investigate the role that CV plays in the removal of a heavy metal (i.e., hexavalent chromium (Cr(VI)) by FeS. The stabilized effect from algal extracellular polymeric substance (EPS) enhanced the reactivity of FeS due to the decrease of FeS aggregation, thus increasing Cr(VI) removal rate from 0.21 min⁻¹ to 0.79 min⁻¹. Furthermore, the strong buffering induced by the algal functional groups could effectively prevent the solution pH from increasing, which improved Cr(VI) removal because acidic solution facilitated Cr(VI) reduction by FeS. However, the complexing capacity from algal EPS made Fe(II) unavailable for Cr(VI) reduction, which led to 35% decrease of Cr(VI) removal. The Fe(II) was oxidized to α-FeOOH by Cr(VI) in the absence of CV, while the unreacted Fe(II) was detected as in the form of Fe(OH)₂ in CV-FeS. Cr(VI) was reduced to Cr(III) and S(-II) was oxidized to elemental sulfur (S₈) regardless of the CV. This work showed the different roles of algae in the removal of Cr(VI) by FeS and provided value information for the application of FeS in the polluted algae-containing water system.

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 complexation with extracellular polymeric substances (EPS) greatly reduces the toxicity of heavy metals towards organisms in the environment. However, the molecular mechanism of EPS−metal complexation remains unclear owing to the limitation of precise analysis for key fractions and functionalities in EPS that associate with metals. Herein, we explored the EPS−Cd (II) complexation by fluorescence excitation emission matrix coupled with parallel factor (EEM−PARAFAC), two-dimensional Fourier transform infrared correlation spectroscopy (2D-FTIR−COS) and X-ray photoelectron spectroscopy (XPS), attempting to explain the mechanisms of EPS in alleviating Cd (II) toxicity toward a green alga Chlorella vulgaris (C. vulgaris). When the algal EPS were removed, the cell internalizations of Cd (II), growth inhibition rate and chlorophyll autofluorescence increased, but the surface adsorption and esterase activities decreased, indicating that the sorption of Cd (II) by EPS was crucial in alleviating the algal toxicity. Moreover, the complexation with proteins in EPS controlled the sorption of Cd (II) to algal EPS, resulting in the chemical static quenching of the proteins fluorescence by 47.69 ± 2.37%. Additionally, the complexing capability of the main functionalities, COO⁻ and C–OH in proteins with Cd (II) was stronger than that of C–O(H) and C–O–C in polysaccharides or C–OH in the humus-related substances. Oxygen atom in protein carboxyl C–O might be the key site of EPS−Cd (II) complexation, supported by the modified Ryan−Weber complexation model and the obvious shift of oxygen valence-electron signal. These findings provide deep insights into understanding the interaction of EPS with heavy metals in aquatic environment.