Performance of different microalgal species in removing nickel and zinc from industrial wastewater

A series of batch experiments was conducted to compare the ability of 11 microalgal species of the same cell density in removing nickel (Ni) and zinc (Zn) from synthetic wastewater. These included Chlorella vulgaris (commercially available), Chlorella sorokiniana and Scenedesmus quadricauda (isolates from polluted water of Wuhan, China), and eight different isolates from Hong Kong. The Wuhan isolate of Scenedesmus removed most Ni, probably due to its large biomass. Nickel concentration was reduced from an initial 30 to 0.9 mg/l after 5 min (97% Ni removal), and further declined to 0.4 mg/l after 90 min of treatment. In wastewater containing 30 mg/l Ni and 30 mg/l Zn, more than 98%, Ni and Zn were removed simultaneously at the end of 5 min treatment, indicating that the presence of Zn in wastewater did not affect Ni removal by this Scenedesmus isolate. The second most effective species for Ni removal was an isolate, tentatively identified as Chlorella miniata, Ni concentration was reduced to 10 mg/l after 90 min, and was only slightly interfered by the presence of Zn. In terms of metal removal per unit biomass or unit surface area of algal cells, C. miniata was the best species in removing Ni and Zn. At the other extreme, one Hong Kong isolate (Synechocystis sp.) did not remove any Ni and only achieved 40% Zn removal. Performance of the other isolates was comparable with the commercial C. vulgaris, less than 50% Ni was removed after 5 h of treatment and Ni removal was significantly reduced by the presence of Zn. All algae tested were found to be viable, showing these 11 species could tolerate a mixture of 30 mg/l Ni and 30 mg/l Zn in wastewater.     

Biodegradation capacity of tributyltin by two Chlorella species

Two microalgal species, Chlorella vulgaris and Chlorella sp., which showed high tributyltin (TBT) tolerant ability were investigated for their capabilities in degrading TBT at sublethal concentration. The distribution of TBT and its degraded products dibutyltin (DBT) and monobutyltin (MBT) in the incubation medium, extracellular surface and intracellular fraction were monitored during an exposure period of 14 days. Results showed that biosorption of TBT by the algal cell wall was the major mechanism in reducing 40% of the initial TBT from the medium in the first 2 days. The half-life of TBT incubated with C. vulgaris was 60 h while that of Chlorella sp. was 80 h. The occurrence of DBT at Day 1 in the culture medium provided direct evidence to the biodegradation of TBT by both Chlorella species. At the end of the experimental period, 27 and 41% of the original TBT were recovered as DBT and MBT in cultures of C. vulgaris, respectively. In contrast, DBT appeared to be the only degradation product of Chlorella sp. and only 26% of the original TBT was transformed to DBT. Despite the same genus, TBT was debutylated to a greater extent to MBT by C. vulgaris, while DBT was the end degradation product by Chlorella sp. The capability of such debutylating process therefore accounted for the higher tolerant ability of C. vulgaris than Chlorella sp.     

Biotoxicity of nickel oxide nanoparticles and bio-remediation by microalgae Chlorella vulgaris

Adverse effects of manufactured nickel oxide nanoparticles on the microalgae Chlorellavulgaris were determined by algal growth-inhibition test and morphological observation via transmission electron microscopy (TEM). Results showed that the NiO nanoparticles had severe impacts on the algae, with 72 h EC(50) values of 32.28 mg NiOL(-1). Under the stress of NiO nanoparticles, C. vulgaris cells showed plasmolysis, cytomembrane breakage and thylakoids disorder. NiO nanoparticles aggregated and deposited in algal culture media. The presence of algal cells accelerated aggregation of nanoparticles. Moreover, about 0.14% ionic Ni was released when NiO NPs were added into seawater. The attachment of aggregates to algal cell surface and the presence of released ionic Ni were likely responsible for the toxic effects. Interestingly, some NiO nanoparticles were reduced to zero valence nickel as determined by X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS) analysis. The maximum ratios of nickel reduction was achieved at 72 h of exposure, in accordance with the time-course of changes in soluble protein content of treated C. vulgaris, implying that some proteins of algae are involved in the process. Our results indicate that the toxicity and bioavailability of NiO nanoparticles to marine algae are reduced by aggregation and reduction of NiO. Thus, marine algae have the potential for usage in nano-pollution bio-remediation in aquatic system.     

二氧化氯杀灭小球藻

实验研究了二氧化氯投加量、小球藻的初始浓度、pH、有机物和氨氮含量对ClO2杀灭来自于水库水的小球藻的影响,考察了ClO2氧化与混凝工艺结合时去除小球藻的效果并对工艺条件进行优化。结果表明,ClO2在投加量1.1 mg/L下,接触10 min,小球藻的杀灭率为71.93%。小球藻的杀灭率随着ClO2投加量的增大和藻初始浓度的升高而提高,随水中有机物含量的增加而显著降低,氨氮含量对小球藻杀灭的效果影响很小。在酸性条件和碱性条件下,小球藻的杀灭率均随pH升高而急剧下降,而在中性至弱碱性区间内,藻的杀灭率随pH升高而缓慢下降。对于某以小球藻为优势藻的供水水库源水,ClO2氧化与混凝工艺结合,藻的去除率高达98.47%。除藻的最佳工艺条件为：二氧化氯投加量为0.5 mg/L,聚合氯化铝为5 mg/L,二氧化氯与混凝剂同时投加。     

Effects of propanil, tebufenozide and mefenacet on growth of four freshwater species of phytoplankton: a microplate bioassay

The Albufera Natural Park situated in Valencia (Spain), with a very rich flora and fauna is surrounded by rice fields in which pesticide spraying is a regular practice. With this in mind, the sensitivity of four algal species, Scenedesmus acutus, Scenedesmus subspicatus, Chlorella vulgaris and Chlorella saccharophila to pesticides propanil, tebufenozide and mefenacet was studied using single species toxicity tests. Organisms were exposed to different concentrations of these herbicides and the algal growth was measured in a microplate reader at 410 nm, at 0, 24, 48 and 72 h. Tebufenozide appeared to be the most inhibitory to Scenedesmus and Chlorella species growth. 72 h EC50 of propanil, tebufenozide and mefenacet ranged from 0.29 to 5.98 mg/l, 0.12 to 0.15 mg/l and from 0.25 to 0.67 mg/l, respectively for the four algal species. The two species of Chlorella were more tolerant than the two species of Scenedesmus.     

Evaluation of different algal species sensitivity to mercury and metolachlor by PAM-fluorometry.

In this study, the pulse-amplitude-modulation (PAM)-fluorometric method was used to evaluate the difference in the sensitivity to mercury (Hg) and metolachlor of six algal species: Ankistrodesmus falcatus, Selenastrum capricornutum, Chlorella vulgaris, Nannoplankton (PLS), Microcystis aeruginosa and Pediastrum biwae. We found that the fluorescence parameters (phiM, the maximal photosystem II (PSII) quantum yield, phi'M, the operational PSII quantum yield at steady state of electron transport, Q(P), the photochemical quenching value, and Q(N), the non-photochemical quenching value) were appropriate indicators for inhibitory effects of mercury but only phi'M and Q(N) were useful for metolachlor. The examined algal species showed very different levels of sensitivity to the effect of Hg and of metolachlor. The most sensitive species to Hg and metolachlor were respectively M. aeruginosa and A. falcatus, while the least sensitive were C. vulgaris and P. biwae. We interpreted these differences by the action mode of pollutants and by the different metabolism properties and morphological characteristics between algal species. These results related to fluorescence parameters may offer useful tool to be used in bioassay for different pollutants. Heterogeneous algal sensitivity to the same pollutant suggests the need to use a battery of species to evaluate the effects of mixtures of pollutants in aquatic systems.     

The comparison of growth and nutrient removal efficiency of Chlorella pyrenoidosa in settled and activated sewage

The microscopic green alga, Chlorella pyrenoidosa was grown in settled and activated sewage under two different culture systems, batch and semi-continuous. Good growth was obtained in both types of wastewater and the algal production was comparable to and even higher than that found in commercial Bristol medium. The semi-continuous culture supported more growth than the batch system. There was a close relationship between algal growth and the amount of nutrient removed from both settled and activated sewage. A more rapid drop in NH(4)(+)-N was found in settle rather than activated sewage. The NH(4)(+)-N of settled sewage dropped from its initial 27 to 5 mg litre(-1) in both culture systems. On the other hand, the NO(3)(-)-N of activated sewage started to decrease from Day 2 onwards and the final NO(3)(-)-N concentration was less than 1 mg litre(-1) (over 90% removal efficiency). The amount of total inorganic nitrogen being reduced due to algal culture was similar in both types of sewage. The changes of phosphate content followed the same trend in both sewage, the P concentration increased slightly in the first two days then decreased, especially in the semi-continuous cultures. The final ortho-P in the sewage treated by Chlorella in semi-continuous culture was less than 5 mg litre(-1) (about 62% reduction). Such removal efficiency was slightly lower than those reported in previous studies. In general, the semi-continuous algal culture appeared to be a more suitable and efficient way for wastewater treatment than the batch system. With respect to the total reduction of wastewater inorganic N and P by means of Chlorella cells, there was no significant difference between settled and activated sewage.     

Wastewater nutrient removal by Chlorella pyrenoidosa and Scenedesmus sp

Two different species of photosynthetic microalgae, Chlorella pyrenoidosa and Scenedesmus sp., were grown in settled and activated sewage filtrates at two different inoculum sizes, aimed to reduce nutrient load from wastewater. Higher growth rates were recorded in cultures with higher inoculum size, and algal cells usually grew better in settled sewage than in activated sewage. As algae started to grow and multiply, both nitrogen and phosphorus content in wastewater decreased significantly. The removal rate was rapid during the first week of growth and more than 2/3 of wastewater N and P was reduced. After the initial period, the removal rate slowed down. At the end of this study, more than 80% of total-P and inorganic N present in settled sewage were reduced but such removal efficiency was lower in activated sewage. In general, high inoculum size of algal cells provided more N and P removal than low inoculum, and Chlorella cells performed better than Scenedesmus. These results suggest that cultivation of Chlorella seems to be one of the feasible methods to reduce the amount of nitrogen and phosphorus entering the nearby coastal water, thus preventing the eutrophication problem. It is also clear that algal ponds with high inoculum size might be more suitable to be installed as a secondary rather than a tertiary treatment process.     

Effect of immobilized microalgal bead concentrations on wastewater nutrient removal

A unicellular green microalga, Chlorella vulgaris, entrapped in calcium alginate as algal beads were employed to remove nutrients (N and P) from simulated settled domestic wastewater. A significantly higher nutrient reduction was found in bioreactors containing algal beads (at concentrations ranging from 4 to 20 beads ml(-1) wastewater) than the blank alginate beads (without algae). A complete removal of NH(4)(+)-N and around 95% reduction of PO(4)(3-)-P was achieved within 24 h of treatment in bioreactors having the optimal algal bead concentration (12 beads ml(-1), equivalent to 1:3 algal beads:wastewater, v/v). The NH(4)(+)-N removal was significantly lower at low (around 4 beads ml(-1)) and high (>15 beads ml(-1)) algal bead concentrations. On the other hand, the effect of bead concentration on phosphate removal was less obvious, and bead concentrations ranging from 8 to 20 beads ml(-1) showed comparable percentages of phosphate reduction. Algal uptake and adsorption on alginate gels were found to be the major processes involved in the removal of N and phosphate in the present study. In addition, NH(4)(+)-N could be lost via ammonia volatilization while PO(4)(3-)-P was removed by chemical precipitation, as alkaline pH was recorded in the immobilized microalgal treatment system.     

Effects of bensulfuron-methyl and cinosulfuron on growth of four freshwater species of phytoplankton

The acute toxicity of sulfonylurea herbicides bensulfuron-methyl and cinosulfuron was tested on the five species of freshwater phytoplankton: Scenedesmus acutus, Scenedesmus subspicatus, Chlorella vulgaris and Chlorella saccharophila. Herbicide concentrations eliciting a 50% growth reduction over 96 h (EC50) ranged from 8 to 104 mg/l for cinosulfuron and from 0.015 to 6.2 mg/l for bensulfuron-methyl. The pesticides bensulfuron-methyl, atrazine and benthiocarb were more toxic than cinosulfuron, chlorsulfuron, molinate, fenitrothion and pyridaphenthion in a toxicity study with the same algal species. The transformation of effective concentrations of bensulfuron-methyl and cinosulfuron and other pesticides, obtained from toxicity measurements, into percent of the saturation level in water is used as a first evaluation of potential hazard to aquatic systems. The herbicides cinosulfuron, methyl-bensulfuron, atrazine and chlorsulfuron were more dangerous than the herbicides benthiocarb and molinate and than the insecticides fenitrothion and pyridaphenthion, in a study of hazard evaluation. The two species of Chlorella were more tolerant to both herbicides than the two species of Scenedesmus. A potential environmental hazard of sulfonylurea herbicides to aquatic systems has to be expected even at low environmental concentrations.     

赤泥复合剂絮凝处理太湖水华藻与海洋赤潮藻

研究了一种新型赤泥复合剂对太湖夏季水华原水、实验室培养海洋赤潮藻及模拟海水的絮凝除藻降浊性能。结果表明,赤泥复合剂具有良好的除藻降浊效果,对太湖水华藻去除率为97.3%,原水浊度去除率为95%;对海洋小球藻和三角褐指藻2种赤潮藻去除率分别高达99.1%和98.9%;对2种模拟海水的最高除浊率分别为96.3%和95.3%,其剩余浊度均低于4 NTU,从而为治理淡水水华与海洋赤潮提供一种新技术,同时为赤泥综合利用提供一种新途径。     

小球藻(Chlorella vulgaris)净化沼液和提纯沼气

为了研究同时提纯沼气和净化沼液的可能,利用沼液作为小球藻(Chlorella vulgaris)的培养基,同时把沼气中的CO2作为小球藻培养的碳源。在无菌培养条件下,得出了最适生长条件为沼液浓度50%、初始pH值6.0、接种小球藻干重为0.20 mg/mL、光质为红光。在此条件下沼液中的COD、TN、TP和BOD5的去除率分别达到了88.5%、91.2%、95.3%和87.6%,沼气中甲烷含量由原先的40%提升到60.2%。很好地同时解决了沼气品质低下和高浓度污染物沼液污染环境的问题。     

Enantioselectivity in toxicity and degradation of dichlorprop-methyl in algal cultures

Enantioselectivity in the toxicity and degradation of the herbicide dichlorprop-methyl (2,4-DCPPM) in algal cultures was studied. Enantioselectivity was clearly observed in the toxicity of racemic 2,4-DCPPM and its two enantiomers. R-2,4-DCPPM showed low toxicity to Chlorella pyrenoidosa and Chlorella vulgaris, but higher toxicity to Scenedesmus obliquus. The observed toxicity was ranked: R-2,4-DCPPM>S-2,4-DCPPM>Rac-2,4-DCPPM; the toxicity of R-2,4-DCPPM was about 8-fold higher than that of Rac-2,4-DCPPM. Additionally, 2,4-DCPPM was quickly degraded, in the initial 12 h, and different algae cultures had different enantioselectivity for the 2,4-DCPPM enantiomers. There was no significant enantioselectivity for 2,4-DCPPM in Chlorella vulgaris in the initial 7 h. However, racemic 2,4-DCPPM was degraded by Scenedesmus obliquus quickly, in the initial 4 h, much quicker, in fact, than the S- or R-enantiomers (racemate>R->S-), indicating that the herbicide 2,4-DCPPM was absorbed enantioselectively by Scenedesmus obliquus. The rapid formation of 2,4-DCPP suggested that 2,4-DCPPM adsorbed by algal cells was catalytically hydrolyzed to the free acid, a toxic metabolite. The production rates of 2,4-DCPP were as follows: Scenedesmus obliquus>Chlorella pyrenoidosa>Chlorella vulgaris, consistent with the degradability of 2,4-DCPPM. Scenedesmus obliquus had quick, but different, degradative and uptake abilities for R-, S-, and Rac-2,4-DCPPM. The R- and S- enantiomers were not hydrolyzed in the first 12 h, while both enantiomers were hydrolyzed slowly after that. These results indicate that some physical and chemical properties of compounds are of importance in determining their enantioselective toxicity and degradation. The ester and its metabolite likely played an important role in enantioselective toxicity to the three algae.     

Zinc acclimation and its effect on the zinc tolerance of Raphidocelis subcapitata and Chlorella vulgaris in laboratory experiments.

The effect of zinc acclimation of Raphidocelis subcapitata (syn. Selenastrum capricornutum) and Chlorella vulgaris on their sensitivity towards this metal was examined in a series of laboratory experiments. These two commonly used algal species were acclimated to 65 microg Zn/l and changes in zinc tolerance were monitored using standard growth inhibition tests. The chemically defined ISO medium was used as a control culture medium. Both species demonstrated a maximum increase in zinc tolerance of a factor of 3 after 100 days of acclimation. Shifts in the shape of the concentration-response curve due to acclimation were observed for R. subcapitata. Compared to non-acclimated algae, acclimated R. subcapitata exhibited higher growth rates in all zinc treatments as well as in the controls. This suggests that the use of ISO-medium results in sub-optimal growth due to zinc deficiency. These effects could not be demonstrated for C. vulgaris. The zinc tolerance of both species decreased significantly one week after returning the acclimated algae to control (ISO) medium. 72hEC50 values based on growth rate were two to four times higher than those calculated using biomass measurements. Algal toxicity test results, particularly if used for metal risk assessments, must not be conducted using nutrient deficient media.     

Effects of zinc on the structure and growth dynamics of a natural freshwater phytoplankton assemblage reared in the laboratory

The response of a natural phytoplankton assemblage to different concentrations of Zn(2+) was evaluated by means of a static laboratory bioassay. Aliquots of surface water, taken in autumn from a non-polluted point of the Reconquista River (Buenos Aires, Argentia), were incubated in mineral nutrient media containing 2.5, 10 or 25 mg litre(-1) of Zn(2+) (as zinc chloride). The comparative structure and dynamics of the communities were followed through periodic physico-chemical and biological analyses of samples taken during 24 days of incubation. Under the experimental conditions of the bioassays, the existence of several Zn(2+) tolerant algal species was shown: the most important of them were Chlorella vulgaris Beij. (Chlorophyceae, Chlorococcales), Nitzschia palea (Kütz.) Smith and Gomphonema parvulum (Kütz.) Kütz. (Bacillariophyceae). It was also demonstrated that the algal responses to Zn(2+) were selectively concentration dependent: at 2.5 and 10 mg litre(-1) a stimulatory effect was observed in the diatoms; at 25 mg litre(-1), diatom toxicity occurred. In contrast, Chlorophyceae growth was stimulated at the maximal Zn level. In general, the diversity, richness and equitability of the community were adversely affected by Zn in a concentration-dependent fashion.     

Removal and reductive dechlorination of triclosan by Chlorella pyrenoidosa

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 800 ng mL⁻¹, more than 50% of triclosan was eliminated by algal uptake from the culture medium during the first 1 h exposure and reached equilibrium after the 6 h treatment. In the biodegradation experiments, a removal percentage of 77.2% was obtained after C. pyrenoidosa was cultivated with 800 ng mL⁻¹ triclosan for 96 h. 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.     

Biological treatment of wastewater by selected aquatic plants

Pollutant-removal efficiency of certain macrophytes and algae, such as Eichhornia crassipes, Microcystis aeruginosa, Scenedesmus falcatus, Chlorella vulgaris and Chlamydomonas mirabilis, has been tested in laboratory conditions to evaluate their potential role in wastewater treatment. Sewage of Varanasi city, mixed with the effluents of about 1200 small-scale industries, was used for the tests. The investigation was performed in three stages i.e. a water hyacinth culture followed by an algal culture, and finally a second water hyacinth culture. For the first water hyacinth culture, 10 water hyacinth plants were grown in a tank of wastewater with 15 days' retention time. In the second stage, algal species were cultured in the treated wastewater for 5 days, whilst in the third stage, water hyacinth plants were again grown for further treatment of the wastewater for 9 days. This three-stage aquaculture resulted in very high reductions of BOD (96.9%), suspended solids (78.1%), total alkalinity (74.6%), PO(4)-P (89.2%), NO(3)-N (81.7%), acidity (73.3%), NH(4)-N (95.1%), COD (77.9%), hardness (68.6%) and coliform bacteria (99.2%). An increase in the concentration of dissolved oxygen (70%) was also observed.     

Population growth of Euchlanis dilatata (Rotifera): combined effects of methyl parathion and food (Chlorella vulgaris)

In the present work, the combined impact of four concentrations (0, 0.0625, 0.125, and 0.25 mg/L) of methyl parathion and three densities (0.5 x 10(6), 1.0 x 10(6), and 2.0 x 10(6) cells/mL) of the green alga Chlorella vulgaris on the population growth of Euchlanis dilatata was studied. In general, regardless of the food level, an increase in the concentration of methyl parathion resulted in a significant reduction of the maximal population density and rate of population increase. The population growth rate in the controls ranged from 0.248 to 0.298; rates were lower in the presence of the pesticide. At any toxicant concentration, rotifers fed higher algal density showed significantly higher population growth compared with those at lower food levels. An interaction between toxicant and food level was evident on the population growth of E. dilatata. Results have been discussed in light of the protective role of algal density on the toxic effects of insecticides on rotifers and the differences in susceptibility to toxicants between planktonic and littoral rotifers.     

二氧化氯杀灭小球藻

实验研究了二氧化氯投加量、小球藻的初始浓度、pH、有机物和氨氮含量对ClO2杀灭来自于水库水的小球藻的影响,考察了ClO2氧化与混凝工艺结合时去除小球藻的效果并对工艺条件进行优化。结果表明,ClO2在投加量1.1 mg/L下,接触10 min,小球藻的杀灭率为71.93%。小球藻的杀灭率随着ClO2投加量的增大和藻初始浓度的升高而提高,随水中有机物含量的增加而显著降低,氨氮含量对小球藻杀灭的效果影响很小。在酸性条件和碱性条件下,小球藻的杀灭率均随pH升高而急剧下降,而在中性至弱碱性区间内,藻的杀灭率随pH升高而缓慢下降。对于某以小球藻为优势藻的供水水库源水,ClO2氧化与混凝工艺结合,藻的去除率高达98.47%。除藻的最佳工艺条件为:二氧化氯投加量为0.5 mg/L,聚合氯化铝为5 mg/L,二氧化氯与混凝剂同时投加。     

Effects of macronutrient additions on nickel uptake and distribution in the dinoflagellate Prorocentrum donghaiense Lu

The influences of macronutrient additions on nickel (Ni) uptake and distribution in the subcellular structures and macromolecular components of the dinoflagellate Prorocentrum donghaiense Lu were examined using a radioisotope tracer method. The results showed that nitrate addition enhanced the uptake of Ni by P. donghaiense, whereas phosphate addition inhibited Ni uptake at high-Ni concentration. Nitrate or phosphate addition significantly affected Ni distribution in the subcellular structures and components. The majority of Ni was found in the soluble substances (>70%) and in the proteins (55.0-79.6%) of the algal cells. Urea reduced the Ni content in the amino acid-carbohydrate but elevated its content in proteins, and shown significantly correlated with the protein content of the algal cells. Thus, nutrient enrichment could influence both metal uptake and its distribution in the subcellular structures and components of the phytoplankton, as well as its subsequent transfer in marine food chains.