The toxicity towards the algal species Pseudokirchneriella subcapitata of 425 organic chemical substances was tested in a growth inhibition test. Precautions were taken to prevent loss of the compounds from the water phase and the test system (closed test system, low biomass, shorter test duration, silanized glass) and to keep pH constant by applying a higher alkalinity. Chemical phase distribution was modelled taking ionization, volatilisation, and adsorption to glass and biomass into consideration. If the modelled water concentration was below 90% of the nominal concentration the calculated EC values were corrected accordingly. The model helped to identify substances, where the calculated water concentration was too uncertain. Substances covering a wide range of physical-chemical properties and different modes of action were tested. Median effect concentrations (EC50) lower than 1000 mg/L were found for 310 substances; 216 of these were in the range from 1 to 1000 mg/L and 94 substances had EC50s below 1 mg/L and should be classified as "Very toxic". 36 substances fell in the group with EC50 > 1000 mg/L. An EC50 could not be established for 79 substances. These 425 different organic substances were tested under uniform conditions and thus considered a valuable source of information for administrators, industry, risk assessors and QSAR modellers.

The prosperity and development of tannery industry have brought about rapid economic growth. However, the tannery landfill without anti-seepage measures in the early stage has generated masses of environmental hazards owing to the lack of awareness in environmental protection. Therefore, it is imperative to pay much attention to the understanding of environmental hazards from tannery waste. In this study, solid samples and groundwater samples were collected from a tannery landfill to study the effect of the characteristic pollutants produced by tanning on chromium distribution with other coexisting substances. The results showed that significant correlations were demonstrated between multiple coexisting substances (total organic carbon, total petroleum hydrocarbons, total nitrogen, Cr, F, Ca, Cu and Pb), indicating the possible same source or they coming from the same tannery production stage. The weights of positive effects and negative effects of coexisting substances on total Cr distribution in the profile decreased in the order: total nitrogen > Cu > Ca > Pb > total organic carbon > F > SO42-> Cd, and Ni > Cl > Hg, respectively. Moreover, the simulation of Visual MINTEQ showed that the cations were mainly bound to Cr as CrO42-, while the anions were bound to Cr3+. This study provided a new perspective on the selection of remediation strategies for Cr-contaminated sites to avoid secondary environmental pollution caused by the release of coexisting heavy metals.

Microplastics (MPs) are ubiquitous in freshwater ecosystems, but knowledge of their effects on extracellular polymeric substance (EPS) produced by algae is poorly understood. The components in specific EPS fractions of Microcystis respond when exposed to MPs is also still unclear. In this study, the responses of Microcystis aeruginosa under polystyrene (PS) microplastic exposure were studied over 17 days of cultivation, using 0.1 μm and 1.0 μm sized PS at three concentration gradients (1, 10 and 100 mg/L). Results indicate that algal growth significantly increased using the 0.1 and 1.0 μm PS at a high concentration (100 mg/L) on day 17, with growth rates of 74.71% ± 0.94% and 35.87% ± 1.23%, respectively. All tested PS had a maximum inhibitory effect on the photosynthesis on day 5, but the inhibition of photosynthetic activity by 0.1 μm PS alleviated after 13 days of exposure, indicating recovery of microalgae from the toxic environment. The two PS sizes at 100 mg/L concentration triggered EPS release in the latter stage of the experiment; meanwhile, fluorescence EEM analysis showed that smaller-sized PS (0.1 μm) at various doses noticeably increased humic acid-like substances in tightly bound EPS (TB-EPS) fractions on day 17. Our findings showed that EPS release and humic acid-like substances secretion of Microcystis likely can resist MPs exposure. The results provide new insights into the toxicity mechanism of MPs on freshwater microalgae, as well as understanding the ecological risks of microplastics.

Organic micropollutants (MPs) pose potential threats to environmental ecosystems and human health. This study investigated uptake of perfluoroalkyl substances (PFASs), pharmaceuticals, and paraben by edible oyster mushrooms (Pleurotus ostreatus), cultivated on spiked growth substrate. Concentrations of pharmaceuticals and paraben in substrate showed a decreasing trend over a 20-day harvesting period, whereas PFAS concentrations were variable over the harvesting period. However, only propylparaben, clarithromycin, and PFASs were detected in fruiting bodies of oyster mushroom. Uptake of PFASs by oyster mushroom fruit bodies was negatively correlated with perfluorocarbon chain length. An impact of MPs on fungal colonization was observed, with decreased respiration in treatments with the highest concentration of MPs, but production of fruiting bodies was not affected by exposure level. The potential human risk from ingestion of MPs was evaluated for oyster mushrooms exposed to the highest concentration of MPs in substrate, based on acceptable daily intake (ADI).

Microbiomes are vital in promoting nutrient cycling and plant growth in soil ecosystems. However, microbiomes face adverse effects from multiple persistent pollutants, including per- and poly-fluoroalkyl substances (PFASs). PFASs threaten the fertility and health of soil ecosystems, yet the response of microbial community stability and trophic transfer efficiencies to PFASs is still poorly understood. This study explored the spatial patterns of PFASs in topsoil environments from the West Taihu Lake Basin of China and links their presence to soil microbial community stability at compositional and functional levels. Our results revealed that PFBA (13.87%), PFTrDA (11.63%), PFDoA (11.02%), PFOA (10.99%), and PFOS (10.39%) contributed the most to the spatial occurrence of PFASs. Soil properties, including salinity (14.47%), uniformity (9.68%), dissolved inorganic carbon (8.62%), and clay content (8.18%), affected PFASs distribution the most. In soil microbiomes, eukaryotic taxa had wider niche breadths and stronger community stability than prokaryotes when exposed to PFASs (p < 0.05). The presence of PFBA and PFHpA inhibited the functional stability of archaeal and bacterial communities (p < 0.05). PFBA and PFPeA reduced the structural stability of heterotrophic bacteria and Myxobacteria, respectively (p < 0.05). Based on null modeling, PFPeA significantly regulated the assembly processes of most microbial sub-communities (p < 0.01). The trophic transferring efficiencies of autotrophic bacteria to metazoan organisms were directly stimulated by PFASs (p < 0.05), and the potential trophic transferring efficiencies of methanogenic archaea to protozoa were inhibited by PFASs (p < 0.05). This study highlighted the potential contributions of PFASs to soil microbial community stability and food webs during ecological soil management.

As pharmaceutical substances are highly used in human and veterinary medicine and subsequently released in the environment, they represent emerging contaminants in the aquatic compartment. Diclofenac (DCF) is one of the most commonly detected pharmaceuticals in water and little research has been focused on its long-term effects on freshwater invertebrates. In this study, we assessed the chronic impacts of DCF on the freshwater gastropod Lymnaea stagnalis using life history, behavioral and molecular approaches. These organisms were exposed from the embryo to the adult stage to three environmentally relevant DCF concentrations (0.1, 2 and 10 μg/L). The results indicated that DCF impaired shell growth and feeding behavior at the juvenile stage, yet no impacts on hatching, locomotion and response to light stress were noted. The molecular findings (metabolomics and transcriptomic) suggested that DCF may disturb the immune system, energy metabolism, osmoregulation and redox balance. In addition, prostaglandin synthesis could potentially be inhibited by DCF exposure. The molecular findings revealed signs of reproduction impairment but this trend was not confirmed by the physiological tests. Combined omics tools provided complementary information and enabled us to gain further insight into DCF effects in freshwater organisms.

Macrolide antibiotics and azole fungicides are detected widely in the aquatic environment as a result of their increased use in humans and animal livestock disease and their incomplete removal by wastewater treatment plants. In most cases, ecotoxicological tests are performed by using individual chemical substances, but because of the coexistence of a number of chemicals in the environment, organisms are exposed to many chemicals simultaneously. Therefore, it is important to evaluate effects of chemical interactions, adding to potential hazards of individual chemical. Here, we investigated the synergetic effects of combined chemicals (the azole fungicide ketoconazole and either of two macrolide antibiotics, erythromycin and clarithromycin) in growth inhibition testing using Pseudokirchneriella subcapitata according to OECD Test guideline 201. Combination index plots, isobolograms, and curve-shift analyses revealed that the combination of macrolide antibiotic and ketoconazole at various ratios resulted in strong synergism that enhanced growth inhibition of P. subcapitata, suggesting the necessity of investigating potential hazard of combined chemicals for regulatory purposes.

Exogenous selenium (Se) improves the tolerance of plants to abiotic stress. However, the effects and mechanisms of different Se species on drought stress alleviation are poorly understood. This study aims to evaluate and compare the different effects and mechanisms of sodium selenate (Na2SeO4) and sodium selenite (Na2SeO3) on the growth, photosynthesis, antioxidant system, osmotic substances and stress-responsive gene expression of Nicotiana tabacum L. under drought stress. The results revealed that drought stress could significantly inhibit growth, whereas both Na2SeO4 and Na2SeO3 could significantly facilitate the growth of N. tabacum under drought stress. However, compared to Na2SeO3, Se application as Na2SeO4 induced a significant increase in the root tip number and number of bifurcations under drought stress. Furthermore, both Na2SeO4 and Na2SeO3 displayed higher levels of photosynthetic pigments, better photosynthesis, and higher concentrations of osmotic substances, antioxidant enzymes, and stress-responsive gene (NtCDPK2, NtP5CS, NtAREB and NtLEA5) expression than drought stress alone. However, the application of Na2SeO4 showed higher expression levels of the NtP5CS and NtAREB genes than Na2SeO3. Both Na2SeO4 and Na2SeO3 alleviated many of the deleterious effects of drought in leaves, which was achieved by reducing stress-induced lipid peroxidation (MDA) and H2O2 content by enhancing the activity of antioxidant enzymes, while Na2SeO4 application showed lower H2O2 and MDA content than Na2SeO3 application. Overall, the results confirm the positive effects of Se application, especially Na2SeO4 application, which is markedly superior to Na2SeO3 in the role of resistance towards abiotic stress in N. tabacum.

The present study investigated the biodegradation of polystyrene (PS) plastic by mealworm (Tenebrio molitor) on different diets followed by untargeted screening of larvae gut intestine tissue and frass (manure and feed residuals) to investigate the existence of polymer-generated organic residues. Three different diets, consisting of PS, rolled barley and water were tested. PS degradation rates ranged from 16% to 23% within 15 days, with no statistical differences in survival rates. The larvae fed with ad libitum barley:PS (20:1 w/w) and water had the highest growth rate, while higher PS consumption was observed for barley:PS of 4:1 w/w. A GC-TOF-MS analysis revealed no contaminating substances in the gut intestine tissue, nor styrene or PS oligomers, whilst several bioactive compounds and traces of alkanes, mostly with small carbon chains, were present. Metabolomics analysis on the collected frass, either on the lipophilic (CHCl3) or the polar fraction (MeOH-H2O) was performed. Styrene and PS oligomers (dimers, trimers) were identified, though in a relatively low total amount, up to a total of 346.0 ng/mg 2,4 di-tert butylphenol was identified in both frass and tissue, coming from the PS polymer (Non-intentionally added substances; NIAS). Finally, in the polar fraction of frass, bioactive molecules (fatty acids, amides) were identified, together with several hydrocarbons, mostly with longer carbon chains. The formation of these substances indicated enzymatic and biochemical activity in the larvae-gut intestine. It was shown that degrading and contaminating organic compounds occur at low levels, in both gut intestine and frass, during bio-degradation of PS.

Phosphorus (P) uptake, distribution and transformation are important processes associated with the growth and metabolism of microalgae. In this study, the fate of P in soluble microbial products (SMP), extracellular polymeric substances (EPS), and intracellular polymeric substances (IPS) of Chlorella vulgaris C7 in the form of inorganic P (IP) and organic P (OP) was studied under different trophic modes, including photoautotrophy, heterotrophy and mixotrophy. The results showed that mixotrophic cultivation of microalgae brought highest biomass yield, which was 2.09 times and 11.10 times higher than that of the photoautotrophic and heterotrophic conditions. Regarding P distribution and transformation, the trophic modes affected the transformation trends and rates of P in the form of IP and OP among SMP, EPS and IPS. Under photoautotrophic condition, EPS was the main P pool, and most of P in C. vulgaris was IP. While, under mixotrophic condition, IPS was the main P pool, and most of P was transformed to OP. The addition of glucose promoted the uptake of P by algal cells and the transformation of IP to OP, which accounted for 79.32% of total phosphorus at the end of cultivation. Collectively, mixotrophic cultivation was regarded as the optimum approach for microalgae cultivation and nutrient recovery.

Acidimicrobiaceae sp. strain A6 (A6), is an anaerobic autotrophic bacterium capable of oxidizing ammonium (NH4+) while reducing ferric iron and is also able to defluorinate PFAS under these growth conditions. A6 is exoelectrogenic and can grow in microbial electrolysis cells (MECs) by using the anode as the electron acceptor in lieu of ferric iron. Therefore, cultures of A6 amended with perfluorooctanoic acid (PFOA) were incubated in MECs to investigate its ability to defluorinate PFAS in such reactors. Results show a significant decrease in PFOA concentration after 18 days of operation, while producing current and removing NH4+. The buildup of fluoride and shorter chain perfluorinated products was detected only in MECs with applied potential, active A6, and amended with PFOA, confirming the biodegradation of PFOA in these systems. This work sets the stage for further studies on the application of A6-based per- and polyfluorinated alkyl substances (PFAS) bioremediation in microbial electrochemical systems for water treatment.

The presence of microplastics in the aquatic ecosystem represents a major issue for the environment and human health. The capacity of organic pollutants to adsorb onto microplastic particles raises additional concerns, as it creates a new route for toxic compounds to enter the food web. Current knowledge on the impact of pristine and/or contaminated microplastics on aquatic organisms remains insufficient, and we provide here new insights by evaluating their biological effects in zebrafish (Danio rerio). Zebrafish larvae were raised in ZEB316 stand-alone housing systems and chronically exposed throughout their development to polyethylene particles of 20-27 μm, pristine (MP) or spiked with benzo[α]pyrene (MP-BaP), supplemented at 1% w/w in the fish diet. While they had no effect at 30 days post-fertilization (dpf), MP and MP-BaP affected growth parameters at 90 and 360 dpf. Relative fecundity, egg morphology, and yolk area were also impaired in zebrafish fed MP-BaP. Zebrafish exposed to experimental diets exhibited an increased incidence of skeletal deformities at 30 dpf as well as an impaired development of caudal fin/scales, and a decreased bone quality at 90 dpf. An intergenerational bone formation impairment was also observed in the offspring of parents exposed to MP or MP-BaP through a reduction of the opercular bone in 6 dpf larvae. Beside a clear effect on bone development, histological analysis of the gut revealed a reduced number of goblet cells in zebrafish fed MP-BaP diet, a sign of intestinal inflammation. Finally, exposure of larvae to MP-BaP up-regulated the expression of genes associated with the BaP response pathway, while negatively impacting the expression of genes involved in oxidative stress. Altogether, these data suggest that long-term exposure to pristine/contaminated microplastics not only jeopardizes fish growth, reproduction performance, and skeletal health, but also causes intergenerational effects.

This study investigated the influence of silver ions (Ag+) on the cytotoxicity of silver nanoparticles (AgNPs) in Phanerochaete chrysosporium and noted the degree of extracellular secretions in response to the toxicant's stress. Oxalate production was elicited with moderate concentrations of 2,4-dichlorophenol (2,4-DCP) and AgNPs reaching a plateau at 10 mg/L and 10 μM, respectively. Increased oxalate accumulation was accompanied by higher activities of manganese peroxidase (MnP) and lignin peroxidase (LiP). However, the secretion of oxalate, MnP and LiP was significantly inhibited owing to Ag+ incorporation into AgNP solution. Production of extracellular polymeric substances (EPS) significantly elevated with an increase in 2,4-DCP concentrations; however, after 24 h of exposure to 100 mg/L 2,4-DCP, an obvious decrease in EPS occurred, indicating that part of EPS could be consumed as carbon and energy sources to ameliorate biological tolerance to toxic stress. Furthermore, AgNP-induced "particle-specific" cytotoxicity was substantially enhanced with additional Ag+ as evidenced by its significant negative impact on cellular growth, plasma membrane integrity, and morphological preservation compared with AgNPs at equal Ag concentration.

Due to the bioaccumulation and non-biodegradability of cadmium, Cd can pose a serious threat to ecosystem even at low concentration. Microalgae is widely distributed photosynthetic organisms in nature, which is a promising heavy metal remover and an effective industrial sewage cleaner. However, there are few detailed reports on the short-term and long-term molecular mechanisms of microalgae under Cd stress. In this study, the adsorption behavior (growth curve, Cd removal efficiency, scanning electron microscope, Fourier transform infrared spectroscopy, and dynamic change of extracellular polymeric substances), cytotoxicity (photosynthetic pigment, MDA, GSH, H2O2, O2-) and stress response mechanism of microalgae were discussed under EC50. RNA-seq detected 1413 DEGs in 4 treatment groups. These genes were related to ribosome, nitrogen metabolism, sulfur transporter, and photosynthesis, and which been proved to be Cd-responsive DEGs. WGCNA (weighted gene co-expression network analysis) revealed two main gene expression patterns, short-term stress (381 genes) and long-term stress (364 genes). The enrichment analysis of DEGs showed that the expression of genes involved in N metabolism, sulfur transporter, and aminoacyl-tRNA biosynthesis were significantly up-regulated. This provided raw material for the synthesis of the important component (cysteine) of metal chelate protein, resistant metalloprotein and transporter (ABC transporter) in the initial stage, which was also the short-term response mechanism. Cd adsorption of the first 15 min was primary dependent on membrane transporter and beforehand accumulated EPS. Simultaneously, the up-regulated glutathione S-transferase (GSTs) family proteins played a role in the initial resistance to exogenous Cd. The damaged photosynthetic system was repaired at the later stage, the expressions of glycolysis and gluconeogenesis were up-regulated, to meet the energy and substances of physiological metabolic activities. The study is the first to provide detailed short-term and long-term genomic information on microalgae responding to Cd stress. Meanwhile, the key genes in this study can be used as potential targets for algae-mediated genetic engineering.

Adhesive biocoating has microstructure composed of biomolecules to entrap viable cells in a stabilized matrix over exposed surfaces. Although marine benthic diatoms are a common group of algae excreting substantial amount of extracellular polymeric substances (EPS), studies regarding the utilization of these EPS are scarce. Using the soluble EPS derived from Navicula incerta and pre-deposition of it as a thin conditioning layer on microporous polyvinylidene fluoride (PVDF) membranes, the pre-coated surface was used to investigate the cell binding affinity of three marine microalgae, namely Amphora coffeaeformis, Cylindrotheca fusiformis and Navicula incerta. Microalgae actively engaged themselves on the pre-coated membranes which was 10 times greater than the initial cell adhesion degree. Soluble EPS is mainly comprised of polysaccharide while bounded EPS is mainly comprised of protein. On EPS pre-coated membranes, N. incerta released the least amount of bounded polysaccharides (<100 mg m-2) and vice versa for the other two because EPS production is usually maximized to assist cell adhesion onto unfavorable substrates. In stark contrast, when the adaptation period (first 6 h) ended, cells began to secrete more bounded protein for cell growth, and an increasing trend of protein content found in N. incerta has verified its optimal adaptation onto the biocoating itself. On pristine PVDF membranes, the adhesion degree was ranked in ascending order: C. fusiformis, N. incerta and A. coffeaeformis. Interestingly, after the pre-coating process, the order was reported as: A. coffeaeformis, N. incerta and C. fusiformis, but it should be noted that C. fusiformis demonstrated fluctuating cell colonization degree and bounded EPS production over time. In other words, the biofilm's susceptibility was confirmed since the cells latched loosely on the membranes rather than in a biofilm matrix. Biocoating enables uniform cell distribution and firmer biofilm growth, opening the door to vast future applications in environmental bioremediation and sensing.

Pipe material and residual chlorine are key factors for the drinking water distribution system, and understanding the biofilm ecosystem is vital for water quality safeguard. The aim of our study was to determine the influence of pipe materials (ductile iron, steel, polyethylene) and chlorination on the biofilm structure and microbial community, as shown by the physicochemical properties, extracellular polymeric substances (EPS) structural characteristics, bacterial community composition, and functional traits. EPS spatial properties were studied based on a semi-quantitative confocal laser scanning microscope (CLSM) description. Regarding the impact of chlorination, residule chlorine (1.0 ± 0.3 mg L-1 free chlorine) could inhibit the bacteria colonization, and initiate a potential response to external disinfectants revealed by the EPS spatial distribution changes and communities variation compared to unchlorinated system. Regarding the impact of pipe material, polyethylene (PE) biofilms displayed lower biomass, loose zoogloea structure, lower proteins and polysaccharides content, and poor microbial diversity in contrast to ductile iron and steel biofilms. Pipe material was the more possible driving factor of the biofilm community composition compared to the chlorination based on principal coordinates analysis (PCoA) and permutational multivariate analysis of variance (PERMANOVA). Actinobacteria was dominant in the PE biofilms (45.57%-83.32%), while Alphaproteobacteria (34.30%-73.22%) and Gammaproteobacteria (6.46%-36.82%) were the major classes in the steel and ductile iron biofilms. The genus Rhodococcus was predominant in the PE biofilms. Rhodococcus, Pseudomonas, and Sphingomonas seemed to have a better growth advantage in the chlorinated system and display a stronger disinfectant resistance. Functional sketch prediction indicated the potential impact of pipe material and chlorination on functional pathway abundnce, possible functional pathways associated with infectious disease included. This study provides insights into the impact of pipe material and chlorination on biofilm structure and microbial community and might help to develop monitoring or maintenance strategies to protect the biosafety of the drinking water.

High salinity seriously inhibits the growth and metabolism of microorganisms, resulting in poor settleability, excessive biomass loss and low treatment efficiency of biological wastewater treatment systems. The development of halophilic aerobic granular sludge (HAGS) is a feasible strategy for addressing this challenge. However, there are problems with the granulation of HAGS and the stability of granules at decreasing temperatures. In this study, granular activated carbon (GAC) with a large specific surface area and good biocompatibility was used to enhance the robustness of HAGS. The results showed that the addition of GAC shortened the granulation time from 60 d (control system) to 35 d (GAC-addition system). The proteins contents of extracellular polymeric substances (EPS) in the GAC-addition system was significantly higher (p < 0.05) than that in the control system during granulation. Satisfactory NH4+-N and chemical oxygen demand (COD) removal efficiencies reached more than 96% in both systems at 18-26 °C. When the operating temperature was lower than 15 °C, the GAC-addition system exhibited better NH4+-N removal performance (>80%) than the control system (<60%). Moreover, the abundance of almost all nitrogen metabolism-related genes in the GAC-addition system was higher than that in the control system. During the granulation process, the enrichment of functional microorganisms, including family Flavobacteriaceae, Rhodobacteraceae, and Cryomorphaceae, may promote the production of EPS by significantly upregulating (p < 0.05) the metabolic pathway "Signaling Molecules and Interaction" in the GAC-addition system. The overexpression of the nitrogen assimilation gene glnA in heterotrophic bacteria (Halomonas and Marinobacterium) may promote the conversion of inorganic nitrogen to extracellular proteins to adapt to the decreased operational temperature. Our findings confirm that GAC addition is a simple but effective strategy to accelerate granulation and enhance the robustness of HAGS in saline wastewater treatment.

Biochar is a black carbon sorbent that has the ability to stabilize organic substances in soil and, therefore, the potential to reduce their bio-availability. This sustainable material can be produced from locally-available agro wastes. The present study, for the first time, investigated the effects of biochars produced from oil palm empty fruit bunches (EFB) and rice husk (RH) on the efficiency of imazapic and imazapyr (two polar members of imidazolinone herbicides) as well as Onduty®, a mixture of them. It was executed in a Malaysian paddy field soil during a 30-day greenhouse experiment. The presence of optimized EFB and RH biochars in the heavy soil generally increases weed seeds germination and plants growth due to stabilization of the herbicides. The effect of EFB biochar was found higher than RH biochar having a higher affinity to the herbicides. An increase in the biochars application rates enhanced their effects as a soil modifier. Differences were more significant in the higher herbicides doses. Efficacies of all the herbicides were generally decreased in the biochar-amended soils. In the presence of 0.5% biochar in soil, the GR50 values for all herbicides were almost similar to biochar-free soil. In a 1.0% biochar-soil mixture, GR50 values of the herbicides increased by about 1.5 times. Bio-efficacies of the herbicides decreased by around 2.0 times as the biochar application rate enhanced to 2.0%. The greatest GR50 values were obtained in the presence of 4.0% biochar in the soils and were about 7.0-8.5 folds, indicating the high capability of amended soil in the stabilization of the herbicides. The findings of this study can help to reduce imidazolinones' pollution and, in this way, prevent the threats of their residues to the environment.