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Energy is a crucial entity for the development and it has various alternative forms of energy sources. Recently, the synthesis of nanoparticles using benign biocatalyst has attracted increased attention. In this study, silver nanoparticles were synthesized and characterized using Azadirachta indica plant-derived phytochemical as the reducing agent. Biomass of the microalga Chlorella sp. cultivated in BG11 medium increased after exposure to low concentrations of up to 0.48 mg L -1 AgNPs. In addition, algal cells treated with 0.24 mg L -1 AgNPs and cultivated in BG11 0 medium which contained no nitrogen source showed the highest hydrogen yield of 10.8 mmol L -1 , whereas the untreated cells under the same conditions showed very low hydrogen yield of 0.003 mmol L -1 . The enhanced hydrogen production observed in the treated cells was consistent with an increase in hydrogenase activity. Treatment of BG11 0 grown cells with low concentration of green synthesized AgNPs at 0.24 mg L -1 enhanced hydrogenase activity with a 5-fold increase of enzyme activity compared to untreated BG11 0 grown cells. In addition, to improve photolytic water splitting efficiency for hydrogen production, cells treated with AgNPs at 0.24 mg L -1 showed highest oxygen evolution signifying improvement in photosynthesis. The silver nanoparticles synthesized using phytochemicals derived from plant enhanced both microalgal biomass and hydrogen production with an added advantage of CO 2 reduction which could be achieved due to an increase in biomass. Hence, treating microalgae with nanoparticles provided a promising strategy to reduce the atmospheric carbon dioxide as well as increasing production of hydrogen as clean energy., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

To reduce the high cost of organic carbon sources in waste resource utilization in the cultivation of microalgae, volatile fatty acids (VFAs) derived from activated sludge were used as the sole carbon source to culture Chlorella sorokiniana under the heterotrophic cultivation. The addition of VFAs in the heterotrophic condition enhanced the total nitrogen (TN) and phosphorus (TP) removal of C. sorokiniana, which proved the advantageous microalgae in using VFAs in the heterotrophic culture after screening in the previous study. To discover the possible mechanism of nitrogen and phosphorus adsorption in heterotrophic conditions by microalgae, the effect of different ratios of VFAs (acetic acid (AA): propionic acid (PA): butyric acid (BA)) on the nutrient removal and growth properties of C. sorokiniana was studied. In the 8:1:1 group, the highest efficiency (77.19%) of VFAs assimilation, the highest biomass (0.80 g L -1 ) and lipid content (31.35%) were achieved, with the highest TN and TP removal efficiencies of 97.44 % and 91.02 %, respectively. Moreover, an aerobic denitrifying bacterium, Pseudomonas, was determined to be the dominant genus under this heterotrophic condition. This suggested that besides nitrate uptake and utilization by C. sorokiniana under the heterotrophy, the conduct of the denitrification process was also the main reason for obtaining high nitrogen removal efficiency., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The strategy of nitrogen sufficiency conversion can improve ammonium nitrogen (NH 4 + -N) removal with microalgal cells from ammonium-rich wastewater. We selected and identified one promising isolated algal strain, NCU-7, Chlorella sorokiniana, which showed a high algal yield and tolerance to ammonium in wastewater, as well as strong adaptability to N deprivation. The transition from N deprivation through mixotrophy (DN, M) to N sufficiency through autotrophy (SN, P) achieved the highest algal yields (optical density = 1.18 and 1.59) and NH 4 + -N removal rates (2.5 and 4.2 mg L -1 d -1 ) from synthetic wastewaters at two NH 4 + -N concentrations (160 and 320 mg L -1 , respectively). Algal cells in DN, M culture obtained the lowest protein content (20.6%) but the highest lipid content (34.0%) among all cultures at the end of the stage 2. After transferring to stage 3, the lowest protein content gradually recovered to almost the same level as SN, P culture on the final day. Transmission electron microscopy and proteomics analysis demonstrated that algal cells had reduced intracellular protein content but accumulated lipids under N deprivation by regulating the reduction in synthesis of protein, carbohydrate, and chloroplast, while enhancing lipid synthesis. After transferring to N sufficiency, algal cells accelerated their growth by recovering protein synthesis, leading to excessive uptake of NH 4 + -N from wastewater. This study provides specific insights into a nitrogen sufficiency conversion strategy to enhance algal growth and NH 4 + -N removal/uptake during microalgae-based ammonium-rich wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

The implementation of industrial-scale facilities for microalgae cultivation is limited due to the high operation costs. One of the main problems in obtaining an efficient and long-lasting microalgae culture system is biofouling. The particular issue when developing antibiofouling surfaces for microalgae cultures is that the material must be transparent. The main purpose of this work was to evaluate the antibiofouling efficiency of a non-toxic polydimethylsiloxane-based coating prepared with polyethylene glycol-based copolymer on different photobioreactors at the pilot-plant scale. The antifouling properties result from the development of a fouling-release coating utilizing hydrogel technology. Nannochloropsis gaditana and Chlorella sorokiniana were cultured outdoors for 3 months over the summer, when biofouling formation is at its highest due to environmental conditions, to test the coating's antibiofouling efficiency. Although biofouling was not completely prevented in either photobioreactor, the coating significantly reduced cell adhesion compared to the polydimethylsiloxane control (70% less adhesion). Therefore, this coating was shown to be a good alternative for constructing efficient closed-photobioreactors at the pilot-plant scale, at least for cultures lasting 3 months., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The algal-bacterial granular sludge (ABGS) system is a prospective wastewater treatment technology, but few studies focused on the effects of different inoculum types on the establishment of the ABGS system under low aeration conditions (step-decrease superficial gas velocity from 1.4 to 0.5 cm/s). Results from this study indicated that compared with other inocula, the ABGS formed by co-inoculating aerobic granular sludge (AGS) and targeted algae (Chlorella) exhibited a shorter granulation period (shortened by 15 days), higher total nitrogen (89.4%) and PO 4 3- -P (95.0%) removal efficiencies, and a greater yield of fatty acid methyl esters (FAMEs) (9.04 mg/g MLSS). This was possibly attributed to that the functional bacteria (e.g. Thauera, Gemmobacter and Rhodobacter) in the inoculated AGS facilitated the ABGS granulation. The inoculated algae promoted their effective enrichment under illumination conditions and enhanced the production of extracellular polymeric substances, thus improving the stability of ABGS. The enriched algae were attached to the outer layer of the granules, which could provide sufficient oxygen for bacterial metabolism, revealing the inherent mechanisms for the good stability of ABGS under low aeration intensity. Overall, the rapid granulation of ABGS can be achieved by inoculating optimal inocula under low aeration conditions, which is convenient and economically feasible, and motivates the application of algal-bacterial consortia., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Incorporation of wastewater from industrial sectors into the design of microalgal biorefineries has significant potential for advancing the practical application of this emerging industry. This study tested various food industrial wastewaters to assess their suitability for microalgal cultivation. Among these wastewaters, defective soy sauce (DSS) and soy sauce wastewater (SWW) were chosen but DSS exhibited the highest nutrient content with 13,500 ppm total nitrogen and 3051 ppm total phosphorus. After diluting DSS by a factor of 50, small-scale cultivation of microalgae was conducted to optimize culture conditions. SWW exhibited optimal growth at 25-30 °C and 300-500 μE m -2 s -1 , while DSS showed optimal growth at 30-35 °C. Based on a 100-mL lab-scale and 3-L outdoor cultivation with an extended cultivation period, DSS outperformed SWW, exhibiting higher final biomass productivity. Additionally, nutrient-concentrated nature of DSS is advantageous for transportation at an industrial scale, leading us to select it as the most promising feedstock for microalgal cultivation. With further optimization, DSS has the potential to serve as an effective microalgal cultivation feedstock for large-scale biomass production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Microalgae such as Chlorella pyrenoidosa, Scenedesmus obliquus and Chlorella sorokiniana were cultivated in domestic wastewater for biohydrogen production. The comparison between the microalgae was executed based on biomass productions, biochemical yields and nutrient removal efficiencies. S. obliquus showed the possibility of growing in domestic wastewater reaching maximum biomass production, lipid, protein, carbohydrate yield and nutrient removal efficiency. All the three microalgae reached high biomass production of 0.90, 0.76 and, 0.71 g/L, respectively for S. obliquus, C. sorokiniana and C. pyrenoidosa. A higher protein content (35.76%) was obtained in S. obliquus. A similar pattern of lipid yield (25.34-26.23%) and carbohydrate yield (30.32-33.21%) was recorded in all selected microalgae. Chlorophyll-a content was higher in synthetic media-grown algae compared algae grown in wastewater. The maximum nutrient removal efficiencies achieved were 85.54% of nitrate by C. sorokiniana, 95.43% of nitrite by C. pyrenoidosa, ∼100% of ammonia and 89.34% of phosphorus by C. sorokiniana. An acid pre-treatment was applied to disintegrate the biomass of microalgae, followed by dark fermentation in batch mode to produce hydrogen. During fermentation process, polysaccharides, protein and lipids were consumed. Maximum hydrogen production of 45.50 ± 0.32 mLH 2 /gVS, 38.43 ± 0.42 mLH 2 /gVS and 34.83 ± 1.82 mL/H 2 /gVS was achieved by C. pyrenoidosa, S. obliquus and C. sorokiniana respectively. Overall, the results revealed the potential of microalgal cultivation in wastewater coupled with maximum biomass production lead to biohydrogen generation for environmental sustainability., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

When the multilayer adsorption of lead (Pb) and fulvic acid (FA) occurs on algal surface, the adsorption capacity of Pb on the algae will increase dramatically, thus increasing the environmental risk of Pb. However, the corresponding mechanism and the influence of environmental factors on the multilayer adsorption remain unclear. Here, microscopic observation methods and batch adsorption experiments were exactly designed to investigate the adsorption behavior of multilayer adsorption of Pb and FA on algal surface. The results of Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) revealed that carboxyl groups were the major functional groups responsible for the binding of Pb ions in multilayer adsorption, and its number was more than that in monolayer adsorption. The solution pH, with an optimal pH of 7, was a critical factor influencing the occurrence of multilayer adsorption because it influences the protonation of the involved functional groups and determines the concentration of Pb 2+ and Pb-FA in the solution. Increasing the temperature was beneficial for multilayer adsorption, with ΔH for Pb and FA varied from +17.12 to +47.68 kJ/mol and +16.19 to +57.74 kJ/mol, respectively. The multilayer adsorption of Pb and FA onto algal surface also followed the pseudo-second order kinetic model, but was extremely slower than the monolayer adsorption of Pb and FA by 30 times and 15 orders of magnitude, respectively. Therefore, the adsorption of Pb and FA in the ternary system had a different adsorption behavior than that in the binary system, which verified the presence of multilayer adsorption of Pb and FA and further support the multilayer adsorption mechanism. This work is important to provide data support for water ecological risk prevention and control of heavy metals., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Co-pollution of microplastics and per- and polyfluoroalkyl substances (PFAS) is prevailing in the aquatic environment. However, the risks of coexisting microplastics and PFAS on organisms remain unknown. This study investigated the response mechanisms of Chlorella sorokiniana (C. sorokiniana) under polystyrene microplastics (PS-MPs) and perfluorooctanoic acid (PFOA) stress, including toxicity and defense mechanisms. C. sorokiniana was exposed to PS-MPs (10 mg/L) and PFOA (0.05, 0.5, and 5 mg/L) and their mixtures for 96 h, respectively. We found that the dominant toxicity mechanism of PFOA and PS-MPs to C. sorokiniana was dissimilar. PS-MPs mainly inhibited photosynthesis through shading effect, while PFOA mainly induced oxidative stress by reactive oxygen species. The co-exposure of PFOA and PS-MPs aggravated biotoxicity (maximum inhibition rate: 27.27 ± 2.44%), such as photosynthesis inhibition, physical damage, and oxidative stress, compared with individuals. To alleviate toxicity, C. sorokiniana activated defense mechanisms. Extracellular polymeric substances were the first barrier to protect cells, the effect on its secretion was ordered PS-MPs+5PFOA > PS-MPs > 5PFOA, and IBRv2 values were 2.37, 1.35, 1.11, respectively. Antioxidant system was thought of second defense pathway, the influence order of treatment groups was PS-MPs+5PFOA > 5PFOA > PS-MPs, and its IBRv2 values were 2.89, 1.69, 0.25, respectively. Our findings provide valuable information on the complex impacts of PFOA and PS-MPs, which facilitate the ecological risk assessment of multiple pollutants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Synthetic phytohormone (SP) is regarded as an attractive candidate for microalgae cultivation due to its potential for high-value microalgae biomass production. Herein, α-naphthylacetic acid (NAA), indomethacin (IN) and 2,4-dichlorophenoxyacetic acid (2,4-D) were used for the mixotrophic cultivation of Chlorella pyrenoidosa with mariculture wastewater (MW) acidogenic fermentation effluent. The growth and lipid accumulation of Chlorella pyrenoidosa added with SP were enhanced, given their high bioavailability of the nutrients. Among these three SPs, IN was optimal for Chlorella pyrenoidosa growth, with the maximum optical density of 1.81. NAA exhibited the best performance for lipid production and the proportion of lipid reached 50.24%. Furthermore, the energy of Chlorella pyrenoidosa cultured with SP preferentially allocated to lipogenesis. To understand the mechanism of lipid accumulation in Chlorella pyrenoidosa in response to SP, the enzyme activities involved in carbon metabolism were determined. The malic enzyme (ME) and acetyl-CoA carboxylase (ACCase) were positively correlated with lipid accumulation. Phosphoenolpyruvate carboxylase (PEPC) was the negative feedback enzyme for lipid synthesis. The findings could provide valuable information for regulation mechanism of lipid accumulation and value-added products recovery by microalgae., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The biological effects and fate of the chiral illicit drug amphetamine in the presence and absence of microplastics on freshwater algae (Chlorella pyrenoids), including acute toxicity, growth inhibition, photosynthetic pigment content, oxidative stress, lipid peroxidation, and enantioselective fate were assessed. An agglomeration and the shading effects of microplastics in algae suspension were also determined. Microplastics were observed to increase the toxicity of amphetamine to algae and reduce algae cell growth. Exposed Chlorella pyrenoids exhibited a reduced algae cell counts in an agglomeration test, wherein algae cells decreased between 18% and 56% among treatment groups exposed to 5-50 mg L -1 of microplastics. The agglomeration test suggested that microplastics might significantly increase the adverse effect on algae. Furthermore, our experiments demonstrated enantioselective degradation of amphetamine in algae, and demonstrated that the S-enantiomer was preferably degraded by algae cells. Adding microplastics to the algae suspension significantly reduced the enantioselectivity, with an EF value of 0.41 compared with amphetamine-alone group (0.34) after 21 d exposure. These results demonstrated the first evidence of microplastics acting as a vehicle to enhance amphetamine toxicity to Chlorella pyrenoids, as well as provided new insights into the co-effect of microplastics and organic contaminants on food source., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

For the sustainable development of the environment, to reduce the high cost and low productivity of microalgae biofuel, nine microalgae strains were screened to study the growh and nutrient removal properties under heterotrophic culture by using the waste carbon source of volatile fatty acids (VFAs). Chlorella sorokiniana (C.sorokiniana) was selected as the best strain with the highest biomass concentration of 0.77 g L -1 , specific growth rate of 0.25 d -1 , biomass productivity of 91.43 mg L -1 d -1 , total nitrogen removal efficiency of 95.96% and total phosphorus removal efficiency of 93.42%. To study the utilization potential of acid-producing wastewater by heterotrophic microalgae, actual acid-producing wastewater was recycled three times for the utilization of C.sorokiniana. After the three utilization cultivation, the removal rates of COD, total nitrogen, ammonia nitrogen, and total phosphorus were 74.44%, 88.05%, 79.08%, and 82.69%, respectively. The total utilization rates of acetic acid, propionic acid, and butyric acid were 58.99%, 70.54%, and 81.52%, respectively. In addition, the highest lipid content of 39.15% and protein content of 42.43% achieved at the third cultivation. After the first cultivation, the composition and diversity of the microbial community structure changed dramatically, with Protebacteria, Bacteroidota, Hydrogenophaga, and Algoriphagus becoming enriched. These results showed a promising way of coupling wastewater treatment with biomass production for long-term sustainability of microalgae lipid production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

A symbiotic system of algae-bacteria, and attached cultivation, are two ways to increase microalgae biomass, and beneficially effect wastewater treatment. However, the possible advantages of the algae-bacteria co-culture in attached cultivation, are still unclear. This paper investigates the effects of different morphologies of a phosphate solubilizing bacteria-Paenibacillus xylanexedens (bacteria supernatant, bacteria, broken bacteria), on the growth of microalgae-Chlorella pyrenoidosa and wastewater treatment in an attached co-culture system. The results show that the broken bacteria had the most significant effect, with the biomass and protein content of Chlorella pyrenoidosa increasing by 125.67% and 25.04%; and the removal rate of COD, NH 4 + -N and PO 4 3- in wastewater increasing by 23.57%, 146.15% and 9.96% respectively. This indicates that the intracellular material of the Paenibacillus xylanexedens was more effective in promoting the biomass growth of Chlorella pyrenoidosa and the removal rates of COD, NH 4 + -N and PO 4 3- , compared to the algae growing without the bacteria. The algae-bacteria symbiotic attached mode was superior to the suspended mode, in terms of both Chlorella pyrenoidosa biomass enhancement and effective wastewater treatment. The addition of different morphologies of Paenibacillus xylanexedens significantly enlarged the difference between the two culture modes. This study provides a new method for coupled algae-bacteria co-cultures for wastewater treatment, based on the symbiotic effect., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The consortium of microalgae and nitrifying bacteria has attracted attention owing to its advantages, such as energy- and cost-efficiency in terms of using only light irradiation without aeration. However, high light intensity can easily cause photoinhibition of nitrifying bacteria, resulting in process breakdown of the consortium. This challenge limits its practical application in outdoor environment. In a previous study, we developed a "light-shielding hydrogel" which entrapped nitrifying bacteria in carbon black-added alginate hydrogel beads and confirmed its effectiveness of protecting the nitrifying bacteria from intense light up to 1600 μmol photons m -2 s -1 . However, the applicability of the light-shielding hydrogel to microalgae-nitrifying bacteria consortia under strong light irradiation has not yet been clarified. In this study, we aimed to establish consortia of Chlorella sorokiniana and nitrifying bacteria immobilised in light-shielding hydrogel and evaluate their nitrification performance under strong light. Three nitrifying bacteria conditions were used: light-shielding hydrogel, hydrogel containing only nitrifying bacteria without carbon black ('hydrogel'), and dispersed nitrifier without immobilisation ('dispersion') as a control. At 1600 μmol photons m -2 s -1 , the dispersion afforded a significant decrease in nitrification activity and subsequent process breakdown. In contrast, light-shielding hydrogel achieved complete nitrification without nitrite accumulation and had nitrification rates of approximately nine and two times higher than those for the dispersion and hydrogel conditions, respectively. Based on the overall evaluation, a possible sequence of process breakdown under strong light was also proposed. This study demonstrated for the first time that the light-shielding hydrogel/consortia combination had potential for applications, which require mitigation of photoinhibition under strong light irradiation. Further, it is expected that the proposed method will contribute to realise the practical application of microalgae-nitrifying bacteria consortia in various countries that experience high sunlight intensity due to their location in the sunbelt areas., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The impact of ionizing radiation on microorganisms such as microalgae is a topic of increasing importance for understanding the dynamics of aquatic ecosystems in response to environmental radiation, and for the development of efficient approaches for bioremediation of mining and nuclear power plants wastewaters. Currently, nothing is known about the effects of ionizing radiation on the microalgal cell wall, which represents the first line of defence against chemical and physical environmental stresses. Using various microscopy, spectroscopy and biochemical techniques we show that the unicellular alga Chlorella sorokiniana elicits a fast response to ionizing radiation. Within one day after irradiation with doses of 1-5 Gy, the fibrilar layer of the cell wall became thicker, the fraction of uronic acids was higher, and the capacity to remove the main reactive product of water radiolysis increased. In addition, the isolated cell wall fraction showed significant binding capacity for Cu 2+ , Mn 2+ , and Cr 3+ . The irradiation further increased the binding capacity for Cu 2+ , which appears to be mainly bound to glucosamine moieties within a chitosan-like polymer in the outer rigid layer of the wall. These results imply that the cell wall represents a dynamic structure that is involved in the protective response of microalgae to ionizing radiation. It appears that microalgae may exhibit a significant control of metal mobility in aquatic ecosystems via biosorption by the cell wall matrix., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The role of dissolved organic matter (DOM) on the biochemical behavior and toxicity of heavy metals in water is very important but complex and unclear. The present work extracted DOM from a natural water and separated it into three fractions, namely humic acid (HA), fulvic acid (FA) and transphilic acid (TPA). Optical detection showed that HA had most aromatic ring skeletons, FA had more aromatic ring hydrophilic groups, and TPA had the largest number of hydroxyl or carboxyl groups. Their effects on the toxicity of Cu by Chlorella pyrenoidosa depended on types and concentration of DOM. In the case of algal exposure to 0.003 mM initial Cu concentration, the final algal optical density increased from 0.317 of the control group to 0.345, 0.645 and 0.435 in the presence of 20, 10 mg L -1 HA, and 10 mg L -1 TPA, respectively, but were suppressed to 0.246, 0.117 and 0.234 in the presence of 10, 20 mg L -1 FA and 20 mg L -1 TPA. Most adsorption isotherms lost the linearity in the presence of HA, FA and TPA. The adsorbed Cu increased from 0.242 to 0.477 mmol g -1 , following the order of increased concentration of HA, FA, and TPA. The formation of ternary complex and the multi-layer adsorption were proposed to explain the significant enhancement adsorption of Cu in the presence of FA and TPA. This study showed that the type and the density of effective functional groups in DOM determined its effects on Cu toxicity and bioavailability to algae., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Calcium decline and cyanobacterial blooms pose a serious threat to the crustacean zooplankton Daphnia, which has a high demand for calcium. In the present study, we exposed two different clones of Daphnia pulex to different combinations of calcium concentrations (0.1, 0.5, 1.0, 5.0, 10.0 mg L -1 ) and food types (100% Chlorella; 80% Chlorella and 20% non-toxic Microcystis; 80% Chlorella and 20% toxic Microcystis) for 16 days, recorded the key life-history traits, and then used an exponential rise function to fit the traits. Results showed toxic Microcystis and low calcium together negatively affected the survival, development, and reproduction of Daphnia. The negative effect of non-toxic Microcystis and low calcium only affected the development and reproduction. The survival time and reproductive performance increased exponentially with increasing calcium concentration and then approached an asymptotic maximum. Both non-toxic and toxic Microcystis reduced the asymptotic maximum of the reproductive performance. The rising rate at which they reached the asymptotes differed significantly among the three food types; i.e., the reproductive performance of Daphnia was affected in a wider range of calcium concentrations under bad food quality. The findings indicated that Microcystis impaired the tolerance of Daphnia to low calcium, which may cause serious consequences in freshwater ecosystems., Competing Interests: Declaration of competing interest The authors declare no actual or potential conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The extreme high CO 2 in industrial exhaust gas cannot be tolerated by microalgae is the key challenge for the application of microalgae in CO 2 bio-sequestration. To provide better insights for this challenge, we chose one high CO 2 tolerant (Chlorella sp. LAMB 31) and non-tolerant (Chlorella sp. LAMB 122) Chlorella sp. to examine their different CO 2 fixation and carbon allocation responses to 40% CO 2 . The results indicated LAMB 31 had a 24-h "lag phase" of biomass increase, during which the transition from PSII-PSI and the increase of lipid synthesis happened to acclimate high CO 2 conditions, followed by the increase of pigments synthesis, carbon fixation rates and polysaccharide productions. However, no acclimating mechanism was observed in LAMB 122, whose biomass, photosynthesis and material synthesis were all gradually collapsed under 40% CO 2 . Finally, four parameters including Chl a, polysaccharides, carbon fixation rates and MDA were selected to be good physiological biomarkers for high CO 2 tolerant strains screenings in the future., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Atrazine is a widely-applied herbicide used primarily to control weeds, which can persist in the ecosystem and exert potential toxicity to phytoplankton in the aquatic environment. In this study, acute toxicity of atrazine on microalgae Chlorella sp. was investigated with different initial cell densities (1 × 10 5 and 1 × 10 6  cells mL -1 ) and exposure periods (4 d and 8 d). Both growth rate and photosynthetic parameters of the microalgae in response of atrazine stress were determined to find out the sensitive indices and toxicological mechanisms. Because of the independence of initial cell density as well as the high sensitivity and reliability, the performance index PI ABS was verified as the most convincing photosynthetic parameter for indicating IC 50 of atrazine on Chlorella sp., being superior to the traditional parameters of growth rate and F V /F M . The IP amplitude (ΔF IP , fluorescence amplitude of the I-to-P-rise in the OJIP curve) was another sensitive biomarker to reflect atrazine stress. Results from chlorophyll fluorescence transient revealed that atrazine damaged the photosystem II (PS II) reaction center, suppressed the electron transport at the donor and receptor sides, and acted on the absorption, transfer, and utilization of light energy. Our results provide confirmatory references for understanding the toxicity and mechanisms of atrazine on freshwater microalgae., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Naphthenic acids (NAs) originate from bitumen and are considered a major contributor to acute toxicity in oil sands process-affected water (OSPW) produced from bitumen extraction processes. To reclaim oil sands tailings and remediate OSPW, in-pit fluid fine tailings can be water-capped as end pit lakes (EPL). Addressing NAs present in OSPW, either through removal, dilution or degradation, is an objective for oil sands reclamation. EPLs can remediate NAs through degradation or dilution or both. To assess and understand degradation potential, Chlorella kessleri and Botryococcus braunii were tested for their tolerance to, and ability to biodegrade, three model NAs (cyclohexanecarboxylic acid, cyclohexaneacetic acid, and cyclohexanebutyric acid). Water sourced from the industry's first EPL, the Base Mine Lake (BML), was used alone as an inoculum or co-cultured with C. kessleri to biodegrade cyclohexanecarboxylic acid and cyclohexanebutyric acid. All cultures metabolized the model compounds via β-oxidation. Biodegradation by the co-culture of C. kessleri and BML inoculum was most effective and rapid: the cyclohexaneacetic acid generated from cyclohexanebutyric acid could be further degraded by the co-culture, while the cyclohexaneacetic acid generated could not be consumed by pure algal cultures or BML inoculum alone. Adding C. kessleri greatly increased the diversity of the microbial community in the BML inoculum; many known hydrocarbon and NA degraders were identified from the 16S rRNA gene sequencing from this co-culture. This more diverse microbial community could have potential for EPL remediation., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Titanium dioxide nanoparticles (TiO 2 NPs) find applications in our day-to-day life because of unique physicochemical properties. Their release into the aquatic environment poses a possible risk to the organisms. However, the continuing exposure of NPs might reduce their bioavailability to marine organisms owing to aggregation and sedimentation in the aqueous systems thus significantly reducing their toxic impact. In this regard, the present study investigates the effect of continuous exposure of TiO 2 NPs to marine microalgae Chlorella sp. under UV-A irradiation through "tanks in series" mode of experiments. In a three-cycle experiment, concentration of TiO 2 NPs in the first cycle was fixed at 62.6 μM, and the interacted nanoparticles was subsequently exposed to fresh batches of algae in the next two cycles. After the interaction, the NPs underwent severe aggregation (mean hydrodynamic diameter 3000 ± 18.2 nm after cycle I) leading to gravitational settling in the medium and thus decreased bioavailability. The aggregation can be attributed to interactions between the particles themselves (homo-aggregation) further aggravated by the presence of the algal cells (hetero-aggregation). Cellular viability after cycle I was found to be only 24.2 ± 2.5%, and it was enhanced to 96.5 ± 2.8% after the cycle III in the course of continuous exposure. The results were validated with estimation of oxidative stress markers such as intracellular ROS (total ROS, superoxide and hydroxyl radicals) and LPO after each cycle of exposure. The continuing decrease in the EPS across the cycles further confirmed the diminishing toxicity of the NPs., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The genus Chlorella contains unicellular green algae that have been used as food supplements. The purpose of this work was to evaluate the safety of the Chlorella sorokiniana strain CK-22 using powdered preparation (CK-22P) both by in vitro and in vivo assays. These included an experiment for cytotoxicity using Chinese hamster lung fibroblasts (V79 cells) and a 13-week repeated-dose oral toxicity trial using Wistar rats. The cytotoxicity was evaluated by MTT assay of a hot water extract (Hw-Ex) and 80% ethanol extract (Et-Ex) of CK-22P, and no effect on cell viability was observed. The 50% viability inhibitory effect (IC50) value for Hw-Ex and Et-Ex were estimated as greater than 73 and 17 μg/ml, respectively. In the subchronic toxicity test, pelleted rodent diet containing 0%, 2.5%, 5% or 10% CK-22P was given to Wistar rats (ten animals/sex/groups) for 13 weeks. During the experimental period, no CK-22P treatment-induced differences in general condition, body weight gain, food and water consumption, ophthalmology, urinalysis, hematology, clinical chemistry, gross pathology, organ weights, histopathology, or animal death were observed. The no-observed-adverse-effect levels (NOAEL) were estimated to be 5.94 g/kg body-weight/day for males and 6.41 g/kg body-weight/day for females., (Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.)

This study evaluated the impact of dissolved organic matter (DOM) of varying molecule weights (MWs) on long-term exposure to Cu and Pb in Chlorella pyrenoidosa. Citric acid, fulvic acid, and humic acid, in the order of increasing MWs, were selected to represent DOM. The results showed that DOM with larger MWs had stronger inhibitory effects on the bioavailability of Cu to algae. However, the biosorption isotherm of Pb in the presence of DOM was different: as Pb equilibrium concentration increased, the biosorption capacity increased sharply to a maximum, then decreased. The maximum values ranged between 0.186 and 0.398 mmol g -1 , as the solution DOM concentration and MW changed, exhibiting a stoichiometric relationship between DOM, Pb and algae. The ternary complex of Pb-DOM-alga formed in a limited Pb concentration range, and increased the percentage of internalized Pb. This research helps to understand the role of DOM in metal uptake in phytoplankton., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Microalgae biomasses are considered promising feedstocks for biofuel and methane productions. Two Continuously Stirred Tank Reactors (CSTR), fed with fresh (CSTR-C) and heat pre-treated (CSTR-T) Chlorella biomass were run in parallel in order to determine methane productions. The methane yield was 1.5 times higher in CSTR-T with regard to CSTR-C. Aiming to understand the microorganism roles within of the reactors, the sludge used as an inoculum (I), plus raw (CSTR-C) and heat pre-treated (CSTR-T) samples were analyzed by high-throughput pyrosequencing. The bacterial communities were dominated by Proteobacteria, Bacteroidetes, Chloroflexi and Firmicutes. Spirochaetae and Actinobacteria were only detected in sample I. Proteobacteria, mainly Alfaproteobacteria, were by far the dominant phylum within of the CSTR-C bioreactor. Many of the sequences retrieved were related to bacteria present in activated sludge treatment plants and they were absent after thermal pre-treatment. Most of the sequences affiliated to the Bacteroidetes were related to uncultured groups. Anaerolineaceae was the sole family found of the Chloroflexi phylum. All of the genera identified of the Firmicutes phylum carried out macromolecule hydrolysis and by-product fermentation. The proteolytic bacteria were prevalent over the saccharolytic microbes. The percentage of the proteolytic genera increased from the inoculum to the CSTR-T sample in a parallel fashion with an available protein increase owing to the high protein content of Chlorella. To relate the taxa identified by high-throughput sequencing to their functional roles remains a future challenge., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Enantioselectivity in ecotoxicity, digestion and uptake of chiral pesticide cyproconazole to Chlorella pyrenoidosa was studied. The 96h-EC50 values of rac- and the four enantiomers were 9.005, 6.616, 8.311, 4.290 and 9.410 mg/L, respectively. At the concentrations of 8 mg/L and 14 mg/L, the contents of pigments exposed in rac-, enantiomer-2 and 4 were higher than that exposed in enantiomer-1 and 3. The superoxide dismutase (SOD) and catalase (CAT) activity of algae exposed to enantiomer-1 and 3 was higher than that exposed to the rac-, enantiomer-2 and 4 at three levels. In addition, the malondialdehyde (MDA) concentrations in algae disposed with enantiomer-1 and 3 were increased remarkably at three levels. For the digestion experiment, the half-lives of four enantiomers in algae suspension were 28.06, 19.10, 21.13, 15.17 days, respectively. During the uptake experiment, the order of the concentrations of cyproconazole in algae cells was enantiomer-4, 2, 3 and 1. Based on these data, we concluded that ecotoxicity, digestion and uptake of chiral pesticide cyproconazole to C. pyrenoidosa were enantioselective, and such enantiomeric differences must be taken into consideration when assessing the risk of cyproconazole to environment., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

In the natural environment, heavy metal contamination can occur as long-term pollution of sites or as pulses of pollutants from wastewater disposal. In this study two freshwater green algae, Chlorella sp. FleB1 and Scenedesmus YaA6, were isolated from lead-polluted water samples and the effects of 24 h vs 4 and 8 d exposure of cultures to lead on growth, photosynthetic physiology and production of reactive oxygen species (ROS) of these algae were investigated. In Chlorella sp. FleB1, there was agreement between lead impacts on chlorophyll content, photosynthesis and growth in most case. However, in Scenedesmus acutus YaA6 growth was inhibited at lower lead concentrations (0.03-0.87 × 10(-9) M), under which ROS, measured by 2',7' dichlorodihydrofluorescein diacetate fluorescence, were 4.5 fold higher than in controls but photosynthesis was not affected, implying that ROS had played a role in the growth inhibition that did not involve direct effects on photosynthesis. Effects of short-term (5 h, 24 h) vs long-term (4 d and 8 d) exposure to lead were also compared between the two algae. The results contribute to our understanding of the mechanisms of lead toxicity to algae., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The present work evaluated the combined effects of cefradine and ceftazidime on the green alga Chlorella pyrenoidosa using response surface methodologies (RSM). After a 48 h-exposure, the population growth rate (PGR), the chlorophyll-a content and the SOD content of the alga increased with increased concentrations of two antibiotics. However, the three responses did not continue to demonstrate significant increases once antibiotic concentrations exceed a moderate level. Three two order polynomial regression equations were obtained to describe well the relationship between the responses of the alga and the two antibiotics' concentration (R(2) = 0.9997, 0.9292 and 0.9039, respectively). Three 3 D-surface graphs and their contour plots showed directly the changing trends of the alga under the combined effects of two antibiotics. This study for the first time employed the RSM in ecotoxicology, which indicated that the RSM should be placed under a feasible and a potential application prospect in toxicity assessment., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Natural and synthetic steroid hormones such as progesterone and norgestrel in the aquatic environment may cause adverse effects on aquatic organisms. This study investigated the biotransformation of progesterone and norgestrel in aqueous solutions by two freshwater microalgae Scenedesmus obliquus and Chlorella pyrenoidosa and elucidated their transformation mechanisms. More than 95% of progesterone was transformed by the two microalgae within 5d. For norgestrel, almost complete transformation by S. obliquus was observed after 5 d, but nearly 40% was remained when incubated with C. pyrenoidosa. The results also showed that these two compounds were not accumulated in the algal cells. Biotransformation was found to be the main mechanism for their loss in the aqueous solutions, and it followed the first-order kinetic model. For progesterone, three main transformation products, i.e. 3β-hydroxy-5α-pregnan-20-one, 3,20-allopregnanedione and 1,4-pregnadiene-3,20-dione, and six minor androgens were identified. For norgestrel, only two transformation products, 4,5-dihydronorgestrel and 6,7-dehydronorgestrel, were identified for the first time. Hydroxylation, reduction and oxidation are proposed to be the main transformation pathways. Among the two microalgae species, S. obliquus was found more efficient in the transformation of the two target compounds than C. pyrenoidosa. The results clearly demonstrated the capability of the two microalgae to transform the two progestogens. The biotransformation and products could have significant environmental implications in the fate and effects of the two steroids., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Cadmium (Cd) shows high toxicity to aquatic microalgae. Many studies showed that Cd inhibited activities of photosystem II (PSII) but the effects of heavy metals on photosystem I (PSI) and cyclic electron flow (CEF) were still controversial and unclear. The effects of CdCl2 on the activities of PSI, PSII and CEF in Chlorella pyrenoidosa was measured simultaneously in the present study. In presence of 200μM of Cd, ultrastructure of some cells was strongly modified. Cd exposure led to decrease of the activities of photosynthetic oxygen evolution and respiration. PSII was more sensitive to Cd treatment than PSI. Cd treatment showed significant inhibition on the photochemical quantum yield and electron transport rate of PSII. Cd increased the quantum yield of non-light-induced non-photochemical fluorescence quenching, indicating the damage of PSII. The activity of PSI showed tolerance to Cd treatment with concentration less than 100μM in the experiment. Linear electron flow (LEF) made significant contribution to the photochemical quantum yield of PSI of the untreated cells, but decreased with increasing Cd concentration. The contribution of CEF to the yield of PSI increased with increasing Cd concentration. The activation of CEF after exposure to Cd played an essential role for the protection of PSI., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The biosorption isotherms for phenanthrene (Phen) by cultured algae, field-collected plankton, and market algae samples (OSs) and their fractions (lipid-LP, lipid free carbon-LF, alkaline nonhydrolyzable carbon-ANHC, and acid nonhydrolyzable carbon-NHC) were established. All the biosorption isotherms are well fitted by the Freundlich model. The biosorption isotherms for the ANHC and NHC fractions are nonlinear and for the other fractions are linear. It was found that the NHC fractions are chemically and structurally different from other fractions by using elemental analysis and Fourier transformed infrared spectroscopy (FTIR), consisting mainly of aliphatic polymethylene carbon. The average KOC values for Phen at Ce=0.005Sw are 10706±2768mLg(-1) and 95843±55817mLg(-1) for the bulk market algal samples and their NHC isolates, respectively. As the NHC fraction for Porphyra contains higher polymethylene carbon than that for Seaweed or Spirulina, it exhibits higher biosorption capacity. Moreover, the logKOC values are significantly higher for the field-collected samples than for the market algae and cultured algae samples. The multivariate correlation shows that the logKOC values are positively related to the LP contents, and negatively to the C/N ratios for the original algal samples. Furthermore, the logKOC values are negatively related to the polarity indices (O/C and O+N/C) for the original samples and their fractions excluding LP fractions. These observations help to understand the role of polarity, LP and NHC fractions, and aliphatic structures in the biosorption of Phen, which requires more attention in the examination of sorption processes in the natural environment., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Triclosan that is widely used as antimicrobial agent has been detected as contaminant in various aquatic environments. In this work, removal and biodegradation of triclosan in water by using a ubiquitous green alga, Chlorella pyrenoidosa was investigated. When C. pyrenoidosa was exposed to a series concentration of triclosan from 100 to 800ngmL(-1), more than 50% of triclosan was eliminated by algal uptake from the culture medium during the first 1h exposure and reached equilibrium after the 6h treatment. In the biodegradation experiments, a removal percentage of 77.2% was obtained after C. pyrenoidosa was cultivated with 800ngmL(-1) triclosan for 96h. A major metabolite from the reductive dechlorination of triclosan was identified by using liquid chromatography coupled with electrospray ionization-mass spectrometry. The ultrastructural morphology of algal cells grown in the presence of triclosan was observed by using transmission electron microscopy and the growth of algal cells was detected. It was found that the trilcosan treatment resulted in the disruption of the chloroplast and the release of organic material into aquatic environment, which indicated that triclosan may affect membrane metabolism., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Protein derived the marine Chlorella ellipsoidea was hydrolyzed using different proteases (papain, trypsin, pepsin and α-chymotrypsin) for production of antioxidative peptide, and the antioxidant activities of their hydrolysates were investigated using free radical scavenging assay by electron spin resonance spin-trapping technique. Among the hydrolysates, the peptic hydrolysate exhibited the highest antioxidant activity compared to other hydrolysates. To identify antioxidant peptide, the peptic hydrolysate was purified using consecutive chromatographic methods, and the antioxidant peptide was identified to be Leu-Asn-Gly-Asp-Val-Trp (702.2 Da) by Q-TOF ESI mass spectroscopy. The antioxidant peptide scavenged peroxyl, DPPH and hydroxyl radicals at the IC(50) values of 0.02, 0.92 and 1.42 mM, respectively. The purified peptide enhanced cell viability against AAPH-induced cytotoxicity on normal cells. Furthermore, the purified peptide reduced the proportion of apoptotic and necrotic cells induced by AAPH, as demonstrated by decreased sub-G(1) hypodiploid cells and decreased apoptotic body formation by flow cytometry., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Present study investigated physiological and biochemical changes in a green alga culture (Chlorella pyrenoidosa) upon exposure to varied concentrations of pentachlorophenol (PCP) (0, 0.4 mgL(-1), 2.5 mgL(-1) and 10 mgL(-1)). The results revealed that with the increase of PCP concentration, a decrease of the algal biomass, levels of photosynthetic pigments (chlorophyll a, b and carotenoid), soluble protein and an increase of nitrate reductase (NR) activity were observed. This suggests that the PCP may serve as an uncoupler, causing low ATP level within the algal cells and led most of the NR molecules to be in the dephosphoration state (i.e. active from). Moreover, it shows that photosynthetic pigments especially carotenoid were more sensitive indicators to indicate PCP toxicity as compared to the other parameters., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

In order to better understand the relationship between lead speciation and its bioavailability in natural waters, the interactions between Pb(II), fulvic acid and the freshwater alga, Chlorella kesslerii were studied at various algal cell densities. An increase in cellular lead or fulvic acid adsorbed to algae was observed with decrease of the cell density from ca. 10(7) to 10(5)cells ml(-1). In the presence of fulvic acid, cellular Pb was greater than that expected for the same free lead ion concentrations in the absence of fulvic acid in agreement to our previous study. This effect was found to be more pronounced at lower cell density, in accordance with increased fulvic acid adsorption to algae. Good fit between experimental observations and model predictions of cellular Pb at various cell densities, was observed by assuming that fulvic acid adsorbed to algae give rise to additional binding sites for Pb(II). The findings of this study indicate that a further extension of the biotic ligand model which includes the formation of a ternary complex and cell density (or concentration) as an input parameter is needed to improve its site-specific predictive capacity, especially in the case of dissolved organic matter-rich surface waters. This extension of predictive capacity would allow to reduce the deviations from the BLM model predictions for microalgae in the presence of dissolved organic matter and hence will serve as a mechanistic tool for establishing ambient site-specific water quality criteria.

The photocatalytic degradation of sulfacetamide, sulfathiazole, sulfamethoxazole and sulfadiazine in water solutions during their illumination of UV radiation (lambda(max) 366 nm) with TiO2 catalyst was examined. The growth-inhibition effect of sulfonamides and intermediate products theirs photodegradation was investigated in aqueous solution with the green alga Chlorella vulgaris. The biodegradability of the investigated compounds was determined in the illuminated solutions and is expressed as Biochemical Oxygen Demand. It was found that all of the investigated sulfonamides in the initial solutions were resistant to biodegradation and were toxic relative to C. vulgaris. The toxicity (EC50 values) relative to C. vulgaris increased in the following order sulfacetamide, sulfathiazole, sulfamethoxazole, sulfadiazine. All of the investigated sulfonamides undergo photocatalytic degradation. The toxicity of intermediate products of the sulfonamides degradation was significantly lower than the toxicity of sulfonamides in the initial solutions and was dependent on illumination time and degradation rate. The intermediate products of photocatalysis in contrast to the initial sulfonamides, might be mineralized using biological methods.

The effective concentrations of copper on the inhibition of the growth of Scenedesmus obliquus, Chlorella pyrenoidosa and Closterium lunula at 96 h (96 h EC50) were determined to be 50, 68 and 200 microg/l, respectively. The low initial bioaccumulation of Cu by C. lunula was found to be responsible for its tolerance to Cu. The amount of Cu accumulated by all three microalgae reached the maximum value and decreased quickly after the peak followed by a slow decrease over the next 6 d. Bioaccumulation of Cu by C. lunula was directly proportional to the initial Cu concentration. After reaching the first peak after 1 d, the bioconcentration factor of Cu by microalgae declined to its minimum value during the exponential growth phase but increased in the stationary growth phase again. This indicates that desorption of Cu from microalgae was higher during the exponential growth phase but lower in the stationary growth phase. Smaller microalgae with low 96 h EC50 values are more efficient in removing Cu from wastewater.

A study was undertaken to examine the uptake of the organophosphate insecticide pyridaphenthion in the chlorophyta Chlorella saccharophila. Algae cultures were exposed to the initial nominal concentration 10.0 mg l(-1) pyridaphention during seven days. The insecticide bioconcentrates in the biomass to the highest level of 441.5 +/- 25.9 mg kg(-1) on the fifth day of exposure and was followed by a decrease to 76.6 +/- 5.1 mg kg(-1) on the seventh day. A model was constructed to describe the dynamic process, which estimated a bioconcentration factor (BCF) equal to 28. The study demonstrates the potential of accumulation of pyridaphenthion in aquatic organisms and helps to expand the pyridaphenthion toxicity database. The replacement of fenitrothion by pyridaphenthion concerning their use in rice flooded cultures is discussed.