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Bacterial associations play a substantial role in large-scale culture and production systems of microalgae which are used in finfish and shellfish hatcheries as live feed and also as products such as algal paste, spray dried and freeze dried algal powders. The present study explores interaction between the marine microalga Isochrysis galbana MBTDCMFRI S002 and its associated bacteria. The algal-bacterial interaction was investigated by co-culturing axenic algal culture with two bacterial symbionts—Alteromonas sp. (MBTDCMFRI Mab 25) and Labrenzia sp. (MBTDCMFRI Mab 26). There was an increase in algal biomass accumulation and growth rate in the presence of added bacterial symbionts which indicates their algal growth promoting role. In addition, our study showed the potential of these bacterial strains to release various growth stimulatory compounds such as antioxidants, siderophores and indole-3-acetic acid which could have a significant positive impact on algal growth. The heterotrophic growth of these bacterial strains on extracellular carbon produced by I. galbana is evident in the present work. Thus, our findings showed that a mutually beneficial associations exists between I. galbana and coexisting bacterial flora which can be further explored to improve productivity and sustainability of aquaculture algal rearing systems.

Background Numerous studies have shown that stress induction and genetic engineering can effectively increase lipid accumulation, but lead to a decrease of growth in the majority of microalgae. We previously found that elevated CO 2 concentration increased lipid productivity as well as growth in Phaeodactylum tricornutum , along with an enhancement of the oxidative pentose phosphate pathway (OPPP) activity. The purpose of this work directed toward the verification of the critical role of glucose-6-phosphate dehydrogenase (G6PDH), the rate-limiting enzyme in the OPPP, in lipid accumulation in P. tricornutum and its simultaneous rapid growth rate under high-CO 2 (0.15%) cultivation. Results In this study, G6PDH was identified as a target for algal strain improvement, wherein G6PDH gene was successfully overexpressed and antisense knockdown in P. tricornutum , and systematic comparisons of the photosynthesis performance, algal growth, lipid content, fatty acid profiles, NADPH production, G6PDH activity and transcriptional abundance were performed. The results showed that, due to the enhanced G6PDH activity, transcriptional abundance and NAPDH production, overexpression of G6PDH accompanied by high-CO 2 cultivation resulted in a much higher of both lipid content and growth in P. tricornutum , while knockdown of G6PDH greatly decreased algal growth as well as lipid accumulation. In addition, the total proportions of saturated and unsaturated fatty acid, especially the polyunsaturated fatty acid eicosapentaenoic acid (EPA; C20:5, n-3), were highly increased in high-CO 2 cultivated G6PDH overexpressed strains. Conclusions The successful of overexpression and antisense knockdown of G6PDH well demonstrated the positive influence of G6PDH on algal growth and lipid accumulation in P. tricornutum . The improvement of algal growth, lipid content as well as polyunsaturated fatty acids in high-CO 2 cultivated G6PDH overexpressed P. tricornutum suggested this G6PDH overexpression-high CO 2 cultivation pattern provides an efficient and economical route for algal strain improvement to develop algal-based biodiesel production.

Co-cultivation systems offer the potential to commercialize microalgae biomass. The key purpose of the study was to understand the relationship between siderophore-producing bacterium Idiomarina loihiensis RS14 and Chlorella variabilis ATCC 12198 strain for Chlorella growth enhancement. After observing growth enhancement in C. variabilis by adding metal chelator deferroxamine mesylate (siderophore standard) and purified siderophore from I. loihiensis (1 mg mL−1), a co-cultivation system was designed where axenic microalgae and co-cultured (microalgae + bacteria) aliquots were grown in (1:9, 9:1, 1:1) volumetric inoculum ratio (mL) under iron-sufficient and iron-deficient conditions. The co-culture volumetric ratio 1:1 (microalgae/bacteria) showed bleaching of microalgae and 1:9 showed less biomass (310 mg L−1) comparatively with 9:1 that increased 35% of biomass, i.e., 650 mg L−1 (axenic) to 1000 mg L−1 (co-cultured) in iron-deficient media. The inoculum ratios were optimized in 100 mL shake flask and 9:1 ratio was further scaled up with the similar conditions, and the co-culture showed 20% increase in biomass, i.e., 285.6 mg L−1 (axenic) to 356 mg L−1 (co-cultured). The co-cultured biomass contains 19.70% lipid content compared with axenic algae, i.e., 18.41% which shows 7% of increase in co-culture. Protein content increased to 30% in co-culture microalgae compared with axenic microalgae. Scanning electron microscope images show crumpled surface of Chlorella cells in co-cultured compared with its axenic cells. This finding is of interest for biofuel production from microalgae, often attained through nutrient-starvation processes leading to oil accumulation.

The toxicity of chemical substances to algal growth is generally measured by the 72–96 h algal growth inhibition test. We have developed a method to assess the toxicity of chemicals in aquatic environments more quickly and simply than conventional testing methods by delayed fluorescence (DF), which reflects the photosynthetic capacity of algae. The DF method is based on a technique for evaluating the amount of change in the decay curve due to the effects of chemicals (ΔDF, DF inhibition). Various studies on DF have been reported; however, few reports have evaluated the decay curve of DF by approach using inductive modeling based on measurement data such as principal component analysis (PCA) and partial least squares regression analysis (PLS). Therefore, the purpose of this study is to examine methods for estimating the magnitude and type of toxicity of chemicals by means of a principal component model (PC model) and multiple regression model (MR model) derived from changes in the decay curves of DF of algae exposed to a wide range of 37 toxic substances that have an effect of clear magnitude on algal growth. The changes in the DF decay curves due to exposure the 37 toxic substances to algae were summarized in the PC model composed of eigenvectors and scores of four principal components. For validation of usefulness, a hierarchical cluster analysis (HCA) of the amount of change in four PC scores revealed that the growth inhibition rate was more influential than the chemical type. We also found the possibility of quantitatively predicting the growth inhibition of chemicals by MR model by the amount of change in the PC scores.

In recent years, heavy rainfall disasters linked to climate change have become more frequent, raising concerns about the release of chemicals stored in factories. Assessing chemical contamination during such emergencies therefore necessitates the development of a quick and easy method for evaluating hazardous contaminants in combination with toxicity testing. This study proposes a “toxicity screening” method that combines biological response testing and chemical analysis to systematically evaluate hazardous contaminants in emergency situations. The toxicity screening method evaluates the water quality in three steps, including water quality measurements and a delayed fluorescence (DF) assay, metal content measurements and a DF assay, and targeted screening analysis and a DF assay. The efficacy of this method was tested using industrial wastewater from 14 locations. Seven of the samples were non-toxic, while the other seven samples were toxic, displaying no observed effect concentration (NOEC) values ranging from 0.625 to 20%. Two toxic samples in the first phase possessed high total chlorine concentrations (0.4 mg L −1 ) and conductivities (2200 mS m −1 ), indicating that the main sources of toxicity were residual chlorine and a high salt concentration. In the second phase, metal content analysis identified metals as the toxicity cause in four samples. In the third phase, the organic contaminants were analyzed, and tri- n -octyl phosphate (TNOP) was detected at a concentration of 0.00027 mg L −1 . The results of solid-phase extraction experiments and exposure tests with TNOP alone indicated that the contribution of TNOP to the toxicity was negligible and that chemicals not adsorbed on the solid-phase extraction cartridges were the cause of toxicity. The proposed method can therefore be considered effective for disaster-related water quality assessment, delivering results within 12 days. Graphical Abstract

Background: Ecological impacts of micro- and nanoplastics particles (MNP) are among the most discussed environmental concerns. In algae, MNP are commonly hypothesized to reduce growth, which is a standard ecotoxicological endpoint. However, the reported test outcomes vary, with both growth inhibition and stimulation being observed. Due to this conflict of information, a data synthesis for MNP potential to cause growth inhibition in toxicity testing is needed. Methods: We performed a meta-analysis study to assess the effect of MNP exposure on algal growth. Twenty studies published between 2010 and 2020 and representing 16 algal species and five polymer materials administered as particles in size range 0.04 to 3000 µm were included in this meta-analysis. A random-effect model was used to estimate the effect size in three datasets: (1) Low concentration range (< 100 mg/L), (2) High concentration range (≥ 100 mg/L), and (3) Full range model (0.004 to 1100 mg/L), which encompassed all studies using the combination of experimental settings (test species, MNP concentration, polymer material, and particle size) yielding the highest effect size within a study. Results: The exposure to MNP was not significantly associated with growth inhibition in any of the models tested. However, a high heterogeneity between the studies was found in all three models. Neither MNP concentration nor polymer material contributed significantly to the heterogeneity, whereas polymer density had a significant moderating effect, with a higher risk of growth inhibition at lower densities. We also identified a publication bias, with small studies that reported significant inhibition being overrepresented in our dataset. Conclusions: The meta-analysis found limited evidence for MNP effect on microalgal growth in the standard algal growth inhibition test. The heterogeneity and varying methodological quality of studies limited the interpretation and the confidence in the findings. For hazard assessment, standardization and controlled exposure are needed as well as more sensitive endpoints that can inform us about the effect mechanisms. Finally, using particle-free controls in such tests cannot account for the presence of inert particulates in the test system, and, hence, does not allow to attribute observed effects to the test polymers.

In this study, algal growth potential tests were performed in water samples collected from six sampling sites in Meiliang Bay, Lake Taihu. The potential release of soluble reactive phosphorus (SRP) by enzymatic hydrolysis of enzymatically hydrolyzable phosphorus (EHP) was simultaneously evaluated. Results show that all studied regions were in highly eutrophic states, with additional nitrogen (N) or phosphorus (P) inputs, inducing negligible further increase in algal growth. EHP in water could be rapidly transformed into SRP, further supporting the proliferation of algal blooms. The shortest EHP mineralization time was calculated as 69 minutes; therefore, limiting specific nutrient inputs alone in extremely eutrophic lakes can have a limited effect on suppressing the proliferation of algal blooms. Methods to establish a suitable environmental fate for excessive nitrogen and phosphorus nutrients may be more effective and provide more significant results.

In Asia, four harmful raphidophyte species, Chattonella malayana , C. marina , C. subsalsa , and C. tenuiplastida , coexist in the tropical waters but only C. marina was detected in temperate waters. This occurrence pattern pointed to a potentially distinct ecophysiological niche occupancy and possible species dispersion. The growth physiology of these species isolated from tropical Southeast Asia was investigated using unialgal cultures in ten temperatures (13.0–35.5°C) and five salinities (15–35) to better understand the factors driving their distribution. The highest maximum specific growth rates were observed in C. subsalsa (0.65 ± 0.01 d -1 ), followed by C. malayana (0.47 ± 0.03 d -1 ), C. marina (0.45 ± 0.02 d -1 ), and C. tenuiplastida (0.39 ± 0.01 d -1 ). Their optimal temperatures were 28.0, 30.5, 25.5, and 30.5°C, respectively, of which C. marina preferred colder water. C. subsalsa exhibited a wider growth temperature range (20.5–35.5°C), followed by C. marina (20.5–30.5°C), C. tenuiplastida (23.0–33.0°C), and C. malayana (25.5–33.0°C). Optimal salinities were similar between C. subsalsa and C. malayana (30), and between C. marina and C. tenuiplastida (25), but C. subsalsa and C. marina exhibited a similar growth salinity range of 15–35, while C. malayana and C. tenuiplastida was 20–35. High values of F v /F m were observed in C. subsalsa and C. marina (> 0.5) in all tested conditions, but F v /F m of C. malayana and C. tenuiplastida were significantly lower at 20.5°C. All four species achieved a maximum cell density of > 10 4 cells mL -1 in their optimal conditions. Optimal temperatures in C. subsalsa and C. marina were identical to previous reports. The high adaptability of C. subsalsa in various temperatures and salinities suggests its high competitiveness and bloom potential. The high adaptability of C. marina in colder waters compared to other species likely contributes to its wide distribution in the temperate Asian waters. The narrow temperature window of C. malayana and C. tenuiplastida suggests their endemicity and limited distribution in the tropical waters. This study provides evidence about the occurrences and bloom potential of Chattonella spp. in Asia, but the endemicity versus dispersion issue remains unresolved.

Dissolved humic substances (DHSs) are the major components of organic matter in the aquatic environment. DHSs are well known to considerably affect the speciation, solubility, and toxicity of a wide variety of pollutants in the aquatic environment. In this study, the effects of the toxicity of heavy metals and hydrophobic organic pollutants (HOPs) on Chlamydomonas reinhardtii in the presence of humic acid (HA) were examined by a microscale algal growth inhibition (μ-AGI) test based on spectrophotometric detection. To clarify the relationship between the chemical properties of HAs and the toxicity change of pollutants, eight HAs from different sources were prepared and used. HAs were responsible for mitigating the toxicity of Hg, Cu, pesticides (γ-HCH, 2,4-D, and DDT), and polycyclic aromatic hydrocarbons (PAHs) such as naphthalene (Nap), anthracene (Ant), and benzo[a]pyrene (BaP). In particular, an approximately 100-fold decrease in the toxicity of BaP was observed in the presence of 10 ppm HAs extracted from tropical peat. The results indicated that the carboxylic group content and the HA molecular weight are correlated to the changes in the heavy metal toxicity. For HOPs, the aromaticity and polarity of HAs are crucial for mitigating their toxicity. Furthermore, it was clearly shown that the lake water including a high concentration of DHSs collected from Central Kalimantan, Indonesia, reduced the toxicity of Hg and γ-HCH on Chlamydomonas reinhardtii . Graphical abstract

Hazard assessment of microplastic is challenging because standard toxicity testing is targeting soluble (at least partially) chemicals. Adverse effects can occur when test organisms are exposed to turbid environments in the presence of various particulate matter (PM), both natural, such as clay and cellulose, and anthropogenic, such as microplastic. It is, therefore, relevant to compare responses to PM exposure between the microplastic and other suspended solids present at ecologically relevant concentrations. This comparison is possible when reference materials are included in the testing of microplastic hazard potential. Here, we evaluated growth inhibition in unicellular alga Raphidocelis subcapitata exposed to different PM (microplastic, kaolin, and cellulose; 10, 100, and 1,000 mg/L); algae without added solids were used as a control. Also, aggregate formation in the exposure systems was analyzed using particle size distribution (PSD) data. At 10–100 mg/L, no adverse growth effects were observed in any treatments; moreover, algal growth was significantly stimulated in kaolin and cellulose treatments compared to the control. However, at 1,000 mg/L, all tested materials exerted growth inhibition, with no significant differences among the materials. Comparing PSD s across the treatments showed that both PM concentration and size of the particle aggregates were significant growth predictors for all materials tested. Therefore, at high concentrations, both natural and anthropogenic materials have a similar capacity to cause growth inhibition. Linking effects in unicellular organisms to microplastic fragments remains a challenge since plastics incorporate chemicals that may leach and elicit specific effects relative to the particulates. The use of reference materials in hazard assessment of plastic litter is needed to delineate these effects.