Acute toxicity of excess mercury on the photosynthetic performance of cyanobacterium, S. platensis--assessment by chlorophyll fluorescence analysis

Measurement of chlorophyll fluorescence has been shown to be a rapid, non-invasive, and reliable method to assess photosynthetic performance in a changing environment. In this study, acute toxicity of excess Hg on the photosynthetic performance of the cyanobacterium S. platensis, was investigated by use of chlorophyll fluorescence analysis after cells were exposed to excess Hg (up to 20 microM) for 2 h. The results determined from the fast fluorescence kinetics showed that Hg induced a significant increase in the proportion of the Q(B)-non-reducing PSII reaction centers. The fluorescence parameters measured under the steady state of photosynthesis demonstrated that the increase of Hg concentration led to a decrease in the maximal efficiency of PSII photochemistry, the efficiency of excitation energy capture by the open PSII reaction centers, and the quantum yield of PSII electron transport. Mercury also resulted in a decrease in the coefficients of photochemical and non-photochemical quenching. Mercury may have an acute toxicity on cyanobacteria by inhibiting the quantum yield of photosynthesis sensitively and rapidly. Such changes occurred before any other visible damages that may be evaluated by other conventional measurements. Our results also demonstrated that chlorophyll fluorescence analysis can be used as a useful physiological tool to assess early stages of change in photosynthetic performance of algae in response to heavy metal pollution.     

Validation of photosynthetic-fluorescence parameters as biomarkers for isoproturon toxic effect on alga Scenedesmus obliquus

Photosynthetic-fluorescence parameters were investigated to be used as valid biomarkers of toxicity when alga Scenedesmus obliquus was exposed to isoproturon [3-(4-isopropylphenyl)-1,1-dimethylurea] effect. Chlorophyll fluorescence induction of algal cells treated with isoproturon showed inactivation of photosystem II (PSII) reaction centers and strong inhibition of PSII electron transport. A linear correlation was found (R2>or=0.861) between the change of cells density affected by isoproturon and the change of effective PSII quantum yield (PhiM'), photochemical quenching (qP) and relative photochemical quenching (qP(rel)) values. The cells density was also linearly dependent (R2=0.838) on the relative unquenched fluorescence parameter (UQF(rel)). Non-linear correlation was found (R2=0.937) only between cells density and the energy transfer efficiency from absorbed light to PSII reaction center (ABS/RC). The order of sensitivity determined by the EC-50% was: UQF(rel)>PhiM'>qP>qP(rel)>ABS/RC. Correlations between cells density and those photosynthetic-fluorescence parameters provide supporting evidence to use them as biomarkers of toxicity for environmental pollutants.     

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.     

Herbicide impact on Hormosira banksii gametes measured by fluorescence and germination bioassays

The innovative bioassay described here involves chlorophyll a fluorescence measurements of gametes from the macroalgae, Hormosira banksii, where gametes (eggs) were exposed to Diuron, Irgarol and Bromacil. Response was assessed as percent inhibition from control of effective quantum yield (DeltaF/Fm') of photosystem II, herein referred to as % PSII Inhibition. This was measured with the dual-channelled pulse amplitude modulated (PAM) fluorometer, ToxY-PAM. The fluorescence bioassay was run simultaneously with an established H. banksii germination bioassay to compare sensitivity, precision, and time-to-result. The fluorescence bioassay gave highly sensitive results evidenced by EC(50)s (% PSII Inhibition) for Diuron, Irgarol and Bromacil being three, four and three orders of magnitude (respectively) lower than EC50s generated from the germination bioassays. Precision of the fluorescence bioassay was demonstrated with low coefficient of variations (<30%) for all three toxicants. With regard to time, the fluorescence bioassay gave results within 6h, as opposed to more than 50h for the germination bioassay.     

Enchytraeidae Fridericia bulbosa as a new test species for soil ecotoxicity assessment

Fridericia bulbosa (Rosa, 1887) was proposed as a new test species to assess soil ecotoxicity. The acute toxicity of mercury (Hg) and bromoxynil (BX) on the mortality of Fridericia bulbosa (F. bulbosa) in OECD soil was investigated. The results suggested there were statistically significant differences (p<0.05) between controls and treatments except lower concentration of exposure to single pollutant. BX was more toxic to F. bulbosa than Hg. The 14 d LC(50) values for F. bulbosa exposed to Hg and BX were 3.87 and 2.41 mg kg(-1), respectively. The effects of mixtures with single pollutants on earthworms were observed differently. Toxicity of Hg on earthworms was significantly influenced by the concentration of BX. BX was the main contributive factor of the combined toxic effects. It can be proved that F. bulbosa is a suitable bio-indicator to measure the acute toxicity of mixed pollutants in soil and the mortality of earthworms may be considered as a valuable and sensitive biomarker to diagnose adverse effect of Hg or BX in soil environment.     

Alteration of chromium effect on photosystem II activity in Chlamydomonas reinhardtii cultures under different synchronized state of the cell cycle

The inhibitory effect of chromium (Cr) on photosystem II (PSII) activity was investigated in the green alga Chlamydomonas reinhardtii during different phases of the cell cycle. Algae were cultivated in continuous light or a light/dark cycle (16:8 h) to obtain a synchronously dividing cell culture. The cell division phases were determined with the DNA-specific fluorescent probe SYBR green using flow cytometry. The effect of Cr on PSII activity was investigated after a 24-h treatment with algal cultures having different proportions of newly divided cells (G₀/G₁), dividing cells at the DNA replication phase (S), and dividing cells at the mitosis phase (G₂/M). Using chlorophyll a fluorescence parameters based on PSII electron transport capacity in dark- (Φ_MII) and light-adapted (Φ′_MII) equilibrium state, we found that the effect of Cr differs depending on the stage of the cell cycle. When algal cultures had a high proportion of cells actively dividing (M phase), the toxic effect of Cr on PSII activity appeared to be much higher and PSII quantum yield was decreased by 80 % compared to algal cultures mainly in the G₀/G₁ phase. Therefore, the inhibitory effect of Cr on photosynthesis appears to be different according to the cell cycle state of the algal population.     

Effect of the herbicide flumioxazin on photosynthetic performance of grapevine (Vitis vinifera L.)

Among the herbicides used in vineyards, the pre-emergence soil-applied flumioxazin (fmx) is a recently used molecule that inhibits chlorophyll biosynthesis in weed species. The aim of this work is to further characterize the effects of fmx on the non-target grapevine (Vitis vinifera L. cv. Chardonnay) using cutting as a model. Several photosynthesis parameters were estimated during 25 days after treatment with various fmx concentrations (from 0.5mM to 50mM). Measuring chlorophyll fluorescence it appeared that fmx or a by-product penetrated the plant throughout roots and spread throughout vessels. Besides the initial target, protox, fmx affected other functions related to photosynthesis. Fmx induced a simultaneous drop of both P(n), g(s) and T. Fmx caused stomatal closure, which partially explains the decrease of the net photosynthesis. The decline in F(v)/F(m) indicates that the photochemistry of PSII and its ability to reduce the primary acceptor Q(A) are also affected by fmx in grapevine. Fmx leads to a decrease in the coefficients of both photochemical and non-photochemical quenching. Simultaneous stomatal closure and decrease in the quantum yield of CO(2) assimilation indicate a change in energy metabolism following fmx stress. After ten days of fmx treatment, analyses of the response of net carbon assimilation in leaves to different intercellular CO(2) concentrations have shown a decrease of the maximum carboxylation velocity of RuBP. Stomatal closure, PSII photochemistry, change in energy metabolism and RuBP activity were affected by fmx treatment.     

Effects of cadmium on the activities of photosystems of Chlorella pyrenoidosa and the protective role of cyclic electron flow

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 200 μM 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 100 μM 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.     

Removal of gaseous elemental mercury by dielectric barrier discharge

The removal of elemental mercury (Hg(0)) with the reactive species produced from dielectric barrier discharge (DBD) was studied. The effects of the operating parameters, such as the applied voltage, residence time, initial concentration and co-existence of other pollutants, were investigated. The removal of Hg(0) was significantly promoted by an increase in the applied voltage of the DBD reactor system. The presence of NO gas decreased the Hg(0) removal efficiency within the range of input powers tested compared to the case of Hg(0)-only due to the competition for ozone between Hg(0) and NO gas in the DBD reactor.     

Brassinosteroid alleviates phenanthrene and pyrene phytotoxicity by increasing detoxification activity and photosynthesis in tomato

The present study was carried out to investigate the effects of exogenously applied 24-epibrassinolide (BR) on growth, gas exchange, chlorophyll fluorescence characteristics, lipid peroxidation and antioxidant systems of tomato seedlings grown under different levels (0, 10, 30, 100 and 300 μM) of phenanthrene (PHE) and pyrene (PYR) in hydroponics. A concentration-dependent decrease in growth, photosynthetic pigment contents, net photosynthetic rate (Pn), stomatal conductance (Gs), maximal quantum yield of PSII (Fv/Fm), effective quantum yield of PSII (Φ_PSII), photochemical quenching coefficient (qP) has been observed following PHE and PYR exposure. By contrast, non-photochemical quenching coefficient (NPQ) was increased. PHE was found to induce higher stress than PYR. However, foliar or root application of BR (50 nM and 5 nM, respectively) alleviated all those depressions with a sharp improvement in the activity of photosynthetic machinery. The activities of guaicol peroxidase (GPOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) as well as content of malondialdehyde (MDA) were increased in a dose-dependent manner under PHE or PYR treatments. Compared with control the highest increments of GPOD, CAT, APX, GR and MDA by PHE/PYR alone treatments were observed following 300 μM concentration, which were 67%, 87%, 53%, 95% and 74% by PHE and 42%, 53%, 30%, 86% and 62% by PYR, respectively. In addition, both reduced glutathione (GSH) and oxidized glutathione (GSSG) were induced by PHE or PYR. Interestingly, BR application in either form further increased enzymatic and non enzymatic antioxidants in tomato roots treated with PHE or PYR. Our results suggest that BR has an anti-stress effect on tomato seedlings contaminated with PHE or PYR and this effect is mainly attributed by increased detoxification activity.     

Temporal variability of mercury speciation in urban air

Semi-continuous measurements of ambient mercury (Hg) species were performed in Detroit, MI, USA for the calendar year 2003. The mean (±standard deviation) concentrations for gaseous elemental mercury (GEM), particulate mercury (HgP), and reactive gaseous mercury (RGM) were 2.2±1.3 ng m⁻³, 20.8±30.0, and 17.7±28.9 pg m⁻³, respectively. A clear seasonality in Hg speciation was observed with GEM and RGM concentrations significantly (p<0.001) greater in warm seasons, while HgP concentrations were greater in cold seasons. The three measured Hg species also exhibited clear diurnal trends which were particularly evident during the summer months. Higher RGM concentrations were observed during the day than at night. Hourly HgP and GEM concentrations exhibited a similar diurnal pattern with both being inversely correlated with RGM. Multivariate analysis coupled with conditional probability function analysis revealed the conditions associated with high Hg concentration episodes, and identified the inter-correlations between speciated Hg concentrations, three common urban air pollutants (sulfur dioxide, ozone, and nitric oxides), and meteorological parameters. This analysis suggests that both local and regional sources were major factors contributing to the observed temporal variations in Hg speciation. Boundary layer dynamics and the seasonal meteorological conditions, including temperature and moisture content, were also important factors affecting Hg variability.     

The effect of mercury on trees and their mycorrhizal fungi

The Oak Ridge Reservation, established in 1942, was the designated site for the construction of the atomic bomb. During a 20-year period from 1944 to 1963 radioactive and toxic chemical pollutants, especially mercury compounds were released into the surrounding waterways. Tree diversity and mycorrhizal presence and abundance were analyzed in the mercury-contaminated floodplains of East Fork Poplar Creek Oak Ridge (EFPC) (Tennessee). A subsequent greenhouse study was conducted to assess the phytotoxic effects of different mercuric solutions on Platanus occidentalis (American Sycamore), inoculated with soils from EFPC. Total soil mercury in the field had no effect on tree diversity. Organic species of mercury proved to be more toxic than inorganic species of mercury and soil inoculants from EFPC had no protective effects against Hg toxicity in our greenhouse study. Comparison of the effects of mercury contamination in our field and greenhouse studies was difficult due to uncontrolled factors.     

Short term recovery of periphyton photosynthesis after pulse exposition to the photosystem II inhibitors atrazine and isoproturon

Aquatic organisms are exposed to fluctuating concentrations of herbicides which contaminate rivers following their use for agricultural or domestic purposes. The development of sensitive bioanalytical tests enabling us not only to detect the effects of those pollutants but to take into account this pattern of exposure should improve the ecological relevance of river toxicity assessment. In this respect, the use of chlorophyll fluorescence measurements is a convenient way to probe the effect of photosystem II (PSII) inhibitors on primary producers. This study was devoted to validate the combined use of two fluorescence parameters, the effective and the optimal quantum yields of PSII photochemistry (Φ_PSII and F_v/F_m), as reliable biomarkers of initial isoproturon (IPU) or atrazine (ATZ) toxicity to natural periphyton in a pulse exposition scenario. Φ_PSII and F_v/F_m were regularly estimated during a 7 h-exposure to each pollutant (0-100 μM) and also later after being transferred in herbicide-free water (up to 36 h). Our results showed that IPU was more toxic than ATZ, but with effects reversible within 12 h. Moreover, these two similarly acting herbicides (i.e. same target site) presented contrasted short term recovery patterns, regarding the previous exposure duration.     

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.     

Metals and arsenic in soils and corresponding vegetation at Central Elbe river floodplains (Germany)

Floodplain soils at the Elbe river are frequently polluted with metals and arsenic. High contents of these pollutants were detected down to subsoil layers. NH4NO3-extractable (phytoavailable) Cd, Ni, and Zn were elevated in horizons with high acidity. Among five common floodplain plant species, Artemisia vulgaris showed highest concentrations of Cd, Cu, and Hg, Alopecurus pratensis of As and Phalaris arundinacea of Ni, Pb, and Zn. Relationships were weak between metal concentrations in plants and phytoavailable stocks in soil. As and Hg uptake seems to be enhanced on long submerged soils. Enrichments of Cd and Hg are linked to a special plant community composition. Grassland herbage sampled in July/August revealed higher concentrations of As (+122%), Hg (+124%), and Pb (+3723%) than in May. To limit harmful transfers into the food chain, low-lying terraces and flood channels revealing highest contaminations or phytoavailabilities should be excluded from mowing and grazing.     

Repeated use of two Chlorella species, C. vulgaris and WW1 for cyclic nickel biosorption.

Two living Chlorella species were used to remove nickel from solution containing 30 micrograms Ni ml-1 in 10 successive cycles. The present study also examined the continued viability of these two algal species after repeated exposure to nickel. The two species of Chlorella were Chlorella vulgaris (commercially available) and WW1 (indigenous species isolated from domestic sewage and was tentatively identified as Chlorella miniata). The nickel removal percentage of WW1 cells was maintained at around 85% in the first five cycles, then declined slightly from the fifth cycle onwards, and finally achieved around 70% removal at the end of the 10th cycle. On the contrary, the removal efficiency of C. vulgaris declined from 50 to 30% during the 10 cycles of nickel bisorption. At the end of these 10 successive cycles, WW1 accumulated a substantial amount of Ni2+ (the cumulative cellular Ni concentration was 0.92% dry w.), while the value was only 0.17% in the case of C. vulgaris. These results suggest that the local isolate, WW1, had more consistent and satisfactory ability for removing Ni than the commercial C. vulgaris. Both algal species were still capable of dividing after each nickel treatment cycle, suggesting that the cells were not killed even when significant amounts of nickel were adsorbed/absorbed. However, Ni exposure adversely affected the physiological activity of algal cells as reflected by the decline in division rate and chlorophyll-a activity in both species. Such negative effects became more obvious as the number of cyclic treatments was increased. Nevertheless, WW1 cells appeared to recover from nickel treatment when re-cultivated in commercial medium for 2 weeks.     

Assessment of the risk posed by the antifouling booster biocides Irgarol 1051 and diuron to freshwater macrophytes

Antifouling paints are used to reduce the attachment of living organisms to the submerged surfaces of ships, boats and aquatic structures, usually by the release of a biocide. Two 'booster' biocides in common use are the triazine herbicide Irgarol 1051 (N-2-methylthio-4-tert-butylamino-6-cyclopropylamino-s-triazine), and diuron (1-(3,4-dichlorophenyl)-3,3-dimethylurea), which are designed to inhibit algal photosynthesis. Previous research has been directed at the effects of these compounds in marine and estuarine environments. In 2001 we sampled the main rivers and shallow freshwater lakes (Broads) of East Anglia UK for Irgarol 1051, its metabolite GS26575 (2-methylamino-4-tert-butylamino-6-amino-s-triazine) and diuron in order to establish the baseline environmental concentrations of these compounds in freshwater systems of eastern UK and to investigate their possible effects on aquatic plants. Irgarol 1051, GS26575 and diuron were found in water samples collected from 21 locations. The highest concentrations were found in the Norfolk and Suffolk Broads in May. The rivers Great Ouse, Wissey, Bure and Yare also contained all three compounds, as did the Great Ouse Cut-off Channel. The toxicity of these biocides to three macrophyte species (Apium nodiflorum, Chara vulgaris, and Myriophyllum spicatum) was investigated. Deleterious effects on relative growth rate, the maximum quantum efficiency (Fv/Fm) of photosystem II and, for Apium, root mass production were found. C. vulgaris was generally most sensitive; growth, especially of roots, was strongly affected in A. nodiflorum; growth rate of M. spicatum was sensitive to diuron. No observed effect concentrations (NOEC) were interpolated using standard toxicological analysis. These were compared with measured environmental concentrations (MEC) to determine the ranges of risk quotients (MEC/NOEC). Both Irgarol 1051 and diuron represented significant risks to A. nodiflorum and C. vulgaris in this area.     

Removal mechanism of elemental mercury by using non-thermal plasma

The removal mechanism of elementary mercury (Hg(0)) by non-thermal plasma (NTP) has been investigated, where dielectric barrier discharge and O(3) injection methods as oxidation techniques are employed, together with the analysis of mercury species deposited on the reactor surface using temperature-programmed desorption and dissociation (TPDD) and scanning electron microscopy-energy dispersive spectroscopy. The removal of Hg(0) by NTP is found to be time-dependent and proceed through three domains; the Hg(0) concentration just slightly decreases as soon as NTP is initiated and then becomes constant for several minutes (Region 1), thereafter starts to decrease rapidly for 1h (Region 2) and, after passing fall-off region, very slowly decreases for about 4h (Region 3). The deposited mercury species on the reactor surface were conglomerated like islands, rather than dispersed uniformly, and their ratio of Hg(0) to O composition is observed to be 1:2. Additionally, the new peak in TPDD spectra observed in the region of 260-380°C is proposed as HgO(3). These results lead us to conclude that the deposited mercury species by NTP have extra O atoms to oxidize the adsorbed Hg(0), resulting in the acceleration of removal rate as the oxidation of Hg(0) proceeds.     

Mercury Concentrations in Lentic Fish Populations Related to Ecosystem and Watershed Characteristics

Predicting mercury (Hg) concentrations of fishes at large spatial scales is a fundamental environmental challenge with the potential to improve human health. In this study, mercury concentrations were examined for five species across 161 lakes and ecosystem, and watershed parameters were investigated as explanatory variables in statistical models. For all species, Hg concentrations were significantly, positively related to wetland coverage. For three species (largemouth bass, pike, and walleye), Hg concentrations were significantly, negatively related to lake trophic state index (TSI), suggestive of growth biodilution. There were no significant relationships between ecosystem size and mercury concentrations. However, Hg concentrations were strongly, positively related to ecosystem size across species. Scores of small or remote lakes that have never been tested could be prioritized for testing using models akin to those presented in this article. Such an approach could also be useful for exploring how Hg concentrations of fishes might respond to natural or anthropogenic changes to ecosystems over time.     

Preparation of mercury emissions inventory for eastern North America

Point and area inventories of anthropogenic mercury emissions documented by US and Canadian environmental agencies have been aggregated into a single archive for analysis and air pollution modeling work. For 5341 point sources and 1634 aggregated area sources, mercury emissions are apportioned among elemental gaseous [Hg(0)], reactive gaseous[Hg(II)], and particulate [Hg(p)] emissions using speciation factors derived from available monitoring measurements. According to this inventory, 4.82 x 10(5) mol of mercury were emitted in calendar year 1996 in the latitude range 24-51 degrees north, and longitude range 64-91 degrees west, which covers most of North America east of the Mississippi River. Using speciation factors consistent with past emission source studies, we find the relative emission proportions among Hg(0):Hg(II):Hg(p) species are 47:35:18. Maps of the various mercury species' emissions patterns are presented. Gridded emission patterns show local mercury emission extremes associated with individual cement production and municipal incineration facilities, and in contrast to past inventories, population centers do not stand out. Considerable uncertainties are still present in estimating emissions from large point sources, as are methods of apportioning emissions among various mercury species.