Effects of ZnWO4 nanoparticles on growth, photosynthesis, and biochemical parameters of the green microalga Raphidocelis subcapitata.

Nanoparticles have applications in many sectors in the society. ZnWO4 nanoparticles (ZnWO4-NPs) have potential in the fabrication of sensors, lasers, and batteries, and in environmental remediation. Thus, these NPs may reach aquatic ecosystems. However, we still do not know their effects on aquatic biota and, to our knowledge, this is the first study that evaluates the toxicity of ZnWO4-NPs in a eukaryotic organism. We evaluated the toxicity of ZnWO4-NPs on the green microalga Raphidocelis subcapitata for 96 h, in terms of growth, cell parameters, photosynthesis, and biochemical analysis. Results show that most of Zn was presented in its particulate form, with low amounts of Zn2+, resulting in toxicity at higher levels. The growth was affected from 8.4 mg L-1, with 96h-IC50 of 23.34 mg L-1. The chlorophyll a (Chl a) content increased at 30.2 mg L-1, while the fluorescence of Chl a (FL3-H) decreased at 15.2 mg L-1. We observed increased ROS levels at 44.4 mg L-1. Regarding photosynthesis, the NPs affected the oxygen evolving complex (OEC) and the efficiency of the photosystem II at 22.9 mg L-1. At 44.4 mg L-1 the qP decreased, indicating closure of reaction centers, probably affecting carbon assimilation, which explains the decay of carbohydrates. There was a decrease of qN (non-regulated energy dissipation, not used in photosynthesis), NPQ (regulated energy dissipation) and Y(NPQ) (regulated energy dissipation via heat), indicating damage to the photoprotection system; and an increase in Y(NO), which is the non-regulated energy dissipation via heat and fluorescence. The results showed that ZnWO4-NPs can affect the growth and physiological and biochemical parameters of the chlorophycean R. subcapitata. Microalgae are the base of aquatic food chains, the toxicity of emerging contaminants on microalgae can affect entire ecosystems. Therefore, our study can provide some help for better protection of aquatic ecosystems.

New Perspective for Macroalgae-Based Animal Feeding in the Context of Challenging Sustainable Food Production.

Food production is facing challenging times due to the pandemic, and climate change. With production expected to double by 2050, there is a need for a new paradigm in sustainable animal feed supply. Seaweeds offer a highly valuable opportunity in this regard. Seaweeds are classified into three categories: brown (Phaeophyceae), red (Rhodophyceae), and green (Chlorophyceae). While they have traditionally been used in aquafeed, their demand in the feed market is growing, parallelly increasing according to the food demand. Additionally, seaweeds are being promoted for their nutritional benefits, which contribute to the health, growth, and performance of animals intended for human consumption. Moreover, seaweeds contain biologically active compounds such as polyunsaturated fatty acids, antioxidants (polyphenols), and pigments (chlorophylls and carotenoids), which possess beneficial properties, including antibacterial, antifungal, antiviral, antioxidant, and anti-inflammatory effects and act as prebiotics. This review offers a new perspective on the valorization of macroalgae biomass due to their nutritional profile and bioactive components, which have the potential to play a crucial role in animal growth and making possible new sources of healthy food ingredients.

Hydrocytium expands the phylogenetic, morphological, and genomic diversity of the poorly known green algal order Chaetopeltidales.

PREMISE: Chaetopeltidales is a small, understudied order of the green algal class Chlorophyceae, that is slowly expanding with the occasional discoveries of novel algae. Here we demonstrate that hitherto unrecognized chaetopeltidaleans also exist among previously described but neglected and misclassified species. METHODS: Strain SAG 40.91 of Characium acuminatum, shown by previous preliminary evidence to have affinities with the orders Oedogoniales, Chaetophorales, and Chaetopeltidales (together constituting the OCC clade), was investigated with light and electron microscopy to characterize its morphology and ultrastructure. Sequence assemblies of the organellar and nuclear genomes were obtained and utilized in bioinformatic and phylogenetic analyses to address the phylogenetic position of the alga and its salient genomic features. RESULTS: The characterization of strain SAG 40.91 and a critical literature review led us to reinstate the forgotten genus Hydrocytium A.Braun 1855, with SAG 40.91 representing its type species, Hydrocytium acuminatum. Independent molecular markers converged on placing H. acuminatum as a deeply diverged lineage of the order Chaetopeltidales, formalized as the new family Hydrocytiaceae. Both chloroplast and mitochondrial genomes shared characteristics with other members of Chaetopeltidales and were bloated by repetitive sequences. Notably, the mitochondrial cox2a gene was transferred into the nuclear genome in the H. acuminatum lineage, independently of the same event in Volvocales. The nuclear genome data from H. acuminatum and from another chaetopeltidalean that was reported by others revealed endogenized viral sequences corresponding to novel members of the phylum Nucleocytoviricota. CONCLUSIONS: The resurrected genus Hydrocytium expands the known diversity of chaetopeltidalean algae and provides the first glimpse into their virosphere.

Bacterial assemblages structure in intensive cultivations of the microalga Tetradesmus obliquus.

The present study shows the characterization of the bacterial communities associated with different systems during the cultivation of the microalga Tetradesmus obliquus. For that, sequential cultivation was performed in three different systems: (1) Photobioreactor bench-scale; (2) flat-panel photobioreactor; and (3) thin-layer cascade. Cultures were monitored daily for growth parameters and biomass samples were collected for characterization of bacterial communities using metagenomic. A total of 195,177 reads were produced, resulting in the identification of 72 OTUs. In the grouping of bacterial communities, 3 phyla, 6 classes, 28 families, and 35 taxa were found. The bacteria Brevundimonas and Porphyrobacter had a higher relative abundance compared with other taxa found. These taxa were present in all cultivation systems forming a possible core community. Bacterial communities associated with different cultivation systems of the microalga T. obliquus showed an increase in taxa richness and diversity in the super-intensive and intensive systems.

An Intensive Study on Algal Diversity in the Ancient Man-Made Aquatic Ecosystem of Tiruvallur, South India: Exploration for Sustainable Development.

Eco-friendly and beneficial nature algae make it prominent in our earth as well as for human life. In recent decades, microalgal applications is sought in varied fields from the remediation of wastes to the production of pharmaceutical products. Still, more extensive research on bioprospecting should to conducted to get the genus-specific or species-specific applications of microalgae with high efficiency. This inquiry was carried out (October 2021 to September 2022) for the effectual understanding of microalgal composition structure along with seasonal physicochemical variations in the age-old holy tank at Tiruvallur, southeast India. This inquiry also acts as the source data and makes the bioprospecting process easier. It also ignites the researchers to address the microalgae seasonal composition structure of peculiar wet environments. A total of 41 microalgae species were recorded, in which six major algal groups were in order of, Chlorophyceae > Bacillariophyceae > Cyanophyceae > Euglenophyceae > Zygnematophyceae > Trebouxiophyceae. Mean seasonal abundance was highest in the summer season (351 cells/L) and lowest in the monsoon (113 cells/L). One-way ANOVA showed seasonal variations of physicochemical parameters, in which the majority of them attained their peak during summer. Mean values of water temperature, pH, salinity, total dissolved solids, total solids, electrical conductivity, chemical oxygen demand, total hardness, total alkalinity, ammonia, nitrite-nitrogen, and nitrate-nitrogen for the summer were 31.43 °C, 8.53, 0.56 ppt, 383 mg/L, 525 mg/L, 0.85 mS/cm, 46.27 mg/L, 300 ppm, 251.67 ppm, 1.51 mg/L, 0.62 mg/L, and 0.70 mg/L, respectively. Karl Pearson's correlation revealed a most significant relationship between water quality factors and algal density. The Shannon's diversity index (2.78-3.39) indicated moderately rich microalgal diversity in the study area. Palmer's pollution index stated that the temple tank was organically polluted all over the study period except November.

Cryopreservation of the Microalgae Scenedesmus sp.

Each phytoplankton species presents a different behavior and tolerance to the cryopreservation process. Therefore, in a species-specific protocol, it is essential to ensure both growth and post-thawing cell viability. In this study, we explored the effect of cryopreservation of Scenedesmus sp. with two cryoprotectants, dimethyl sulfoxide (DMSO) and methanol (MET), at 5% and 10% inclusion for each. In the control treatment, the microalgae were not exposed to cryoprotective agents (Control). Three post-thawing cell viability criteria were used: no cell damage (NCD), cell damage (CD), and marked lesions (LM), and mitochondrial and cell membrane damage was evaluated by flow cytometry. The study was a 2 × 2 factorial design, with five replications by treatments, population growth, and cell damage evaluated from the fifth day after thawing. On the fifth day, the highest percentage of NCD was observed when the microalgae were cryopreserved with DMSO 5% (50%); Regarding the control group, it showed 0% NCD. Flow cytometry analysis reveals minor damage at the membrane and mitochondria (9-10.7%) when DMSO is used at both inclusion percentages (5-10%) after thawing. In the exponential phase, the highest growth rates, doubling time, and yield was observed in cryopreserved cells with MET 5%. The results suggest that DMSO 5% is an ideal treatment for cryopreserving microalgae Scenedesmus sp.

Genus Ulothrix (Chlorophyceae) in the State of São Paulo, Brazil / Gênero Ulothrix (Chlorophyceae) no Estado de São Paulo, Brasil

Floristic survey of genus Ulothrix (Ulotrichaceae, Chlorophyceae) in the State of São Paulo, southeast Brazil. Five species were identified based on their cell morphology, described in detail, and illustrated. Ulothrix tenerrima and U. tenuissima occurred in four municipalities each, U. aequalis var. aequalis f. aequalis in three municipalities, U. zonata in two municipalities, and U. subtilissima in a single municipality; U. tenuissima being presently cited for the first time for the State of São Paulo.

Levantamento florístico do gênero Ulothrix (Ulotrichaceae, Chlorophyceae) no Estado de São Paulo, sudeste do Brasil. Foram identificadas cinco espécies com base em sua morfologia celular externa e interna, descritas em detalhes e ilustradas. Ulothrix tenerrima e U. tenuissima ocorreram em quatro municípios cada uma, U. aequalis var. aequalis f. aequalis em três, U. zonata em dois, e U. subtilissima apenas em um município; U. tenuissima foi presentemente citada pela primeira vez para o Estado de São Paulo.

Testing for terrestrial and freshwater microalgae productivity under elevated CO2 conditions and nutrient limitation.

BACKGROUND: Microalgae CO2 fixation results in the production of biomass rich in high-valuable products, such as fatty acids and carotenoids. Enhanced productivity of valuable compounds can be achieved through the microalgae's ability to capture CO2 efficiently from sources of high CO2 contents, but it depends on the species. Culture collections of microalgae offer a wide variety of defined strains. However, an inadequate understanding of which groups of microalgae and from which habitats they originate offer high productivity under increased CO2 concentrations hampers exploiting microalgae as a sustainable source in the bioeconomy. RESULTS: A large variety of 81 defined algal strains, including new green algal isolates from various terrestrial environments, were studied for their growth under atmospheres with CO2 levels of 5-25% in air. They were from a pool of 200 strains that had been pre-selected for phylogenetic diversity and high productivity under ambient CO2. Green algae from terrestrial environments exhibited enhanced growth up to 25% CO2. In contrast, in unicellular red algae and stramenopile algae, which originated through the endosymbiotic uptake of a red algal cell, growth at CO2 concentrations above 5% was suppressed. While terrestrial stramenopile algae generally tolerated such CO2 concentrations, their counterparts from marine phytoplankton did not. The tests of four new strains in liquid culture revealed enhanced biomass and chlorophyll production under elevated CO2 levels. The 15% CO2 aeration increased their total carotenoid and fatty acid contents, which were further stimulated when combined with the starvation of macronutrients, i.e., less with phosphate and more with nitrogen-depleted culture media. CONCLUSION: Green algae originating from terrestrial environments, Chlorophyceae and Trebouxiophyceae, exhibit enhanced productivity of carotenoids and fatty acids under elevated CO2 concentrations. This ability supports the economic and sustainable production of valuable compounds from these microalgae using inexpensive sources of high CO2 concentrations, such as industrial exhaust fumes.

A concise review of the bioactivity and pharmacological properties of the genus Codium (Bryopsidales, Chlorophyta).

The genus Codium is one of the most important genera of marine green macroalgae. Its distribution is widespread worldwide and it has a high degree of diversity in species and characteristics. This genus plays an important ecological role in marine ecosystems as it is a primary producer. However, some species in the genus Codium are invasive species and may disturb the functioning of the ecosystem. Economically, Codium has promising potential as a source of diverse nutritional and pharmacological compounds. Codium is edible, has a high nutrient value, and is rich in bioactive compounds. Hence, some species of Codium have been consumed as food and used as herbal medicines in some Asian countries. In recent decades, studies of the bioactivity and pharmacological properties of the genus Codium have attracted the attention of scientists. This review aims to identify gaps in studies analyzing Codium that have been conducted in the past three decades by assessing published research articles on its bioactivity and pharmacological properties. Compounds obtained from Codium have demonstrated significant biological activities, such as immunostimulatory, anticoagulant, anticancer, anti-inflammatory, antioxidant, antiviral, antibacterial, antifungal, antitumor, anti-angiogenic, osteoprotective, and anti-obesity activities. This review provides information that can be used as a future guideline for sustainably utilizing the genus Codium.

A Fast-Growing Oleaginous Strain of Coelastrella Capable of Astaxanthin and Canthaxanthin Accumulation in Phototrophy and Heterotrophy.

Considering the importance of microalgae as a promising feedstock for the production of both low- and high-value products, such as lipids and pigments, it is desirable to isolate strains which simultaneously accumulate these two types of products and grow in various conditions in order to widen their biotechnological applicability. A novel freshwater strain from the genus Coelastrella was isolated in Belgium. Compared to other Coelastrella species, the isolate presented rapid growth in phototrophy, dividing 3.5 times per day at a light intensity of 400 µmol·m-2·s-1 and 5% CO2. In addition, nitrogen depletion was associated with the accumulation of astaxanthin, canthaxanthin, and fatty acids, which reached ~30% of dry weight, and a majority of SFAs and MUFAs, which are good precursors for biodiesel. This strain also accumulated astaxanthin and canthaxanthin in heterotrophy. Although the content was very low in this latter condition, it is an interesting feature considering the biotechnological potential of the microalgal heterotrophic growth. Thus, due to its rapid growth in the light, its carotenogenesis, and its fatty acids characteristics, the newly identified Coelastrella strain could be considered as a potential candidate for biorefinery purposes of both low- and high-values products.

Evaluation of the effects of input variables on the growth of two microalgae classes during wastewater treatment.

Wastewater treatment carried out by microalgae is usually affected by the type of algal strain and the combination of cultivation parameters provided during the process. Every microalga strain has a different tolerance level towards cultivation parameters, including temperature, pH, light intensity, CO2 content, initial inoculum level, pretreatment method, reactor type and nutrient concentration in wastewater. Therefore, it is vital to supply the right combination of cultivation parameters to increase the wastewater treatment efficiency and biomass productivity of different microalgae classes. In the current investigation, the decision tree was used to analyse the dataset of class Trebouxiophyceae and Chlorophyceae. Various combinations of cultivation parameters were determined to enhance their performance in wastewater treatment. Nine combinations of cultivation parameters leading to high biomass production and eleven combinations each for high nitrogen removal efficiency and high phosphorus removal efficiency for class Trebouxiophyceae were detected by decision tree models. Similarly, eleven combinations for high biomass production, nine for high nitrogen removal efficiency, and eight for high phosphorus removal efficiency were detected for class Chlorophyceae. The results obtained through decision tree analysis can provide the optimum conditions of cultivation parameters, saving time in designing new experiments for treating wastewater at a large scale.

Phenolic compounds and antioxidant capacity of Pediastrum boryanum (Chlorococcales) biomass.

Bioactive compounds, synthesized by photosynthetic microorganisms, have drawn the attention of the pharmaceutical field. This study aimed at evaluating synthesis and in vitro antioxidant capacity of phenolic compounds produced by a microalgae species P. boryanum, which was grown in six different culture media (standard BG11, modified BG11/MBG11, standard WC, modified WC, WC*2 and basal). The highest concentrations of biomass (1.75 ± 0.01 g.L-1) and phenolic content (3.18 ± 0.00 mg.g-1) were obtained when P. boryanum was grown in MBG11 and phenolic acids were identified: gallic, protocatechuic, chlorogenic, hydroxybenzoic and vanillic ones. All extracts exhibited scavenger activity in the ABTS assay and inhibited peroxidase. However, phenolic compounds from P. boryanum grown in BG11 and MBG11 had the most potent scavenger activity in the DPPH assay. In sum, P. boryanum can be a new source of free phenolic compounds with potential antioxidant activity when grown in MBG11, since it yields high amounts of biomass and phenolic compounds.

Toxicity of α-Ag2WO4 microcrystals to freshwater microalga Raphidocelis subcapitata at cellular and population levels.

Silver-based materials have microbicidal action, photocatalytic activity and electronic properties. The increase in manufacturing and consumption of these compounds, given their wide functionality and application, is a source of contamination to freshwater ecosystems and causes toxicity to aquatic biota. Therefore, for the first time, we evaluated the toxicity of the silver tungstate (α-Ag2WO4), in different morphologies (cube and rod), for the microalga Raphidocelis subcapitata. To investigate the toxicity, we evaluated the growth rate, cell complexity and size, reactive oxygen species (ROS) production and chlorophyll a (Chl a) fluorescence. The α-Ag2WO4 - R (rod) was 1.7 times more toxic than α-Ag2WO4-C (cube), with IC10 and IC50 values of, respectively, 8.68 ± 0.91 µg L-1 and 13.72 ± 1.48 µg L-1 for α-Ag2WO4 - R and 18.60 ± 1.61 µg L-1 and 23.47 ± 1.16 µg L-1 for α-Ag2WO4-C. The release of silver ions was quantified and indicated that the silver ions dissolution from the α-Ag2WO4 - R ranged from 34 to 71%, while the Ag ions from the α-Ag2WO4-C varied from 35 to 97%. The α-Ag2WO4-C induced, after 24 h exposure, the increase of ROS at the lowest concentrations (8.81 and 19.32 µg L-1), whereas the α-Ag2WO4 - R significantly induced ROS production at 96 h at the highest concentration (31.76 µg L-1). Both microcrystal shapes significantly altered the cellular complexity and decreased the Chl a fluorescence at all tested concentrations. We conclude that the different morphologies of α-Ag2WO4 negatively affect the microalga and are important sources of silver ions leading to harmful consequences to the aquatic ecosystem.

Structural variation and evolution of chloroplast tRNAs in green algae.

As one of the important groups of the core Chlorophyta (Green algae), Chlorophyceae plays an important role in the evolution of plants. As a carrier of amino acids, tRNA plays an indispensable role in life activities. However, the structural variation of chloroplast tRNA and its evolutionary characteristics in Chlorophyta species have not been well studied. In this study, we analyzed the chloroplast genome tRNAs of 14 species in five categories in the green algae. We found that the number of chloroplasts tRNAs of Chlorophyceae is maintained between 28-32, and the length of the gene sequence ranges from 71 nt to 91 nt. There are 23-27 anticodon types of tRNAs, and some tRNAs have missing anticodons that are compensated for by other types of anticodons of that tRNA. In addition, three tRNAs were found to contain introns in the anti-codon loop of the tRNA, but the analysis scored poorly and it is presumed that these introns are not functional. After multiple sequence alignment, the Ψ-loop is the most conserved structural unit in the tRNA secondary structure, containing mostly U-U-C-x-A-x-U conserved sequences. The number of transitions in tRNA is higher than the number of transversions. In the replication loss analysis, it was found that green algal chloroplast tRNAs may have undergone substantial gene loss during the course of evolution. Based on the constructed phylogenetic tree, mutations were found to accompany the evolution of the Green algae chloroplast tRNA. Moreover, chloroplast tRNAs of Chlorophyceae are consistent with those of monocotyledons and gymnosperms in terms of evolutionary patterns, sharing a common multi-phylogenetic pattern and rooted in a rich common ancestor. Sequence alignment and systematic analysis of tRNA in chloroplast genome of Chlorophyceae, clarified the characteristics and rules of tRNA changes, which will promote the evolutionary relationship of tRNA and the origin and evolution of chloroplast.

A comparative test of the gamete dynamics theory for the evolution of anisogamy in Bryopsidales green algae.

Gamete dynamics theory proposes that anisogamy arises by disruptive selection for gamete numbers versus gamete size and predicts that female/male gamete size (anisogamy ratio) increases with adult size and complexity. Evidence has been that in volvocine green algae, the anisogamy ratio correlates positively with haploid colony size. However, green algae show notable exceptions. We focus on Bryopsidales green algae. While some taxa have a diplontic life cycle in which a diploid adult (=fully grown) stage arises directly from the zygote, many taxa have a haplodiplontic life cycle in which haploid adults develop indirectly: the zygote first develops into a diploid adult (sporophyte) which later undergoes meiosis and releases zoospores, each growing into a haploid adult gametophyte. Our comparative analyses suggest that, as theory predicts: (i) male gametes are minimized, (ii) female gamete sizes vary, probably optimized by number versus survival as zygotes, and (iii) the anisogamy ratio correlates positively with diploid (but not haploid) stage complexity. However, there was no correlation between the anisogamy ratio and diploid adult stage size. Increased environmental severity (water depth) appears to drive increased diploid adult stage complexity and anisogamy ratio: gamete dynamics theory correctly predicts that anisogamy evolves with the (diploid) stage directly provisioned by the zygote.

Factors controlling phytoplankton dynamics in an arid reservoir in Tunisia (case of Sidi Saad dam).

Temporal variation of the phytoplankton community and environmental factors were investigated over 8 months: from January 2018 to November 2018 in the Sidi Saad reservoir (central Tunisia): May, June (spring season), July, August (summer season) September, October and November (autumn season), and January (winter season). The relationships between phytoplankton and environmental factors were explored using Canonical Correspondence Analysis (CCA). General linear models (GLMs) were used to predict the phytoplankton abundance. Analysis of variance (ANOVA) was used to test the hypothesis that the abundance of each group of taxa differed between sampling months. The results of chemical analysis of the reservoir showed that the environment was enriched in nitrates, nitrites, ammonium, and orthophosphate, especially in May. Carlson's trophic index using average chemical variables showed that Sidi Saad has a mesotrophic statute. There is temporal phytoplankton succession in the Sidi Saad reservoir. Cyanophyceae dominated over the year except in winter and late autumn (November and January). Chlorophyceae was the dominant group in winter month. The CCA results showed that six environmental factors, orthophosphates (PO43-), total phosphates (TP), Secchi disc depth (SD), total nitrogen (TN), temperature (T), and nitrite (NO2-) had significant influences on the changes in phytoplankton. GLM showed that PO43-, TP, TN, SD, and T were the significant predictors of phytoplankton abundance. Phytoplankton composition is largely dominated by the species Microcystis aeruginosa which formed a bloom with excessive abundance (up to 89.76 billion cell l-1 in spring). We recommend banning the fishing and their consumption during the period of Microcystis bloom and installing a system of biomonitoring of the levels of toxins in the water.

Long-term ecotoxicological effects of ciprofloxacin in combination with caffeine on the microalga Raphidocelis subcapitata.

Ciprofloxacin (CIP) is an antimicrobial "pseudo-persistent" in aquatic ecosystems. Once dispersed in the water compartments, it can also affect the microalgae. Thus, the evaluation of its long-term ecotoxicological effects is necessary. CIP interactions with other pharmaceuticals are not well known. In this study, we investigated the toxic effects of CIP alone and combined with caffeine (CAF), using the modified Gompertz model parameters and the chlorophyll-a production of the microalga Raphidocelis subcapitata as endpoints, throughout a 16-day exposure assay. The exposure to CIP alone led to significant reductions of the growth rate and the cell density of the microalgae compared to control groups. The combination with CAF lowered the adverse effects of CIP to R. subcapitata. However, as the toxicity is dynamic, our results indicated that the toxic effects in respect to the studied endpoints changed throughout the exposure period, reinforcing the need for longer-term ecotoxicity assessments.

A scientometric analysis of ecotoxicological studies with the herbicide atrazine and microalgae and cyanobacteria as test organisms.

Atrazine (ATZ) is one of the most widely used herbicides in the world. A scientometric study was conducted to analyze the evolution of research on ATZ. The study also looked at the use of microalgae and cyanobacteria as biological models for toxicity tests during the period from 1959 to 2019, in the category of toxicology of Web of Science. The results show an increase in the number of scientific publications mainly in the USA, Canada, and China. The majority of papers was published in journals with high impact factors, demonstrating the relevance of such studies. About 83% of the studies aimed to evaluate the effect of ATZ on non-target organisms. From those, 7.5% included microalgae and cyanobacteria. The majority of them worked with chlorophyceae to perform toxicity bioassays of ATZ and analyze its sublethal effects. The gaps identified by this analysis included a small number of collaborations between research groups from different countries; the number of studies with terrestrial organisms, which are larger in comparison to aquatic organisms; and the fact that none of the studies with ATZ and microalgae was performed in the field. These findings can point out to researchers and funding agencies the gaps in knowledge on the toxic effects of ATZ and guide the development of new research projects as well as environmental policies.

Using multiple endpoints to assess the toxicity of cadmium and cobalt for chlorophycean Raphidocelis subcapitata.

Metals may cause damage to the biota of contaminated environments. Moreover, using multiple endpoints in ecotoxicological studies is useful to better elucidate the mechanisms of toxicity of these compounds. Therefore, this study aimed to evaluate the effects of cadmium (Cd) and cobalt (Co) on growth, biochemical and photosynthetic parameters of the microalgae Raphidocelis subcapitata, through quantification of lipid classes composition, chlorophyll a (Chl a) content, maximum (ΦM) and effective (Φ'M) quantum yields and efficiency of the oxygen-evolving complex (OEC). Both metals affected the algal population growth, with an IC50-96h of 0.67 and 1.53 µM of Cd and Co, respectively. Moreover, the metals led to an increase in the total lipid content and reduced efficiency of OEC and ΦM. Cell density was the most sensitive endpoint to detect Cd toxicity after 96 h of treatment. Regarding Co, the photosynthetic parameters were the most affected and the total lipid content was the most sensitive endpoint as it was altered by the exposure to this metal in all concentrations. Cd led to increased contents of the lipid class wax esters (0.89 µM) and phospholipids (PL - at 0.89 and 1.11 µM) and decreased values of triglycerides (at 0.22 µM) and acetone-mobile polar lipids (AMPL - at 0.44 and 1.11 µM). The percentage of free fatty acids (FFA) and PL of microalgae exposed to Co increased, whereas AMPL decreased in all concentrations tested. We were able to detect differences between the toxicity mechanisms of each metal, especially how Co interferes in the microalgae at a biochemical level. Furthermore, to the best of our knowledge, this is the first study reporting Co effects in lipid classes of a freshwater Chlorophyceae. The damage caused by Cd and Co may reach higher trophic levels, causing potential damage to the aquatic communities as microalgae are primary producers and the base of the food chain.