Cylindrospermopsis raciborskii exudate-Cu complexes: impact on copper dynamics and bioavailability in an aquatic food chain.

INTRODUCTION: The increasing contamination of aquatic environments motivates studies on the interactions among natural dissolved organic matter, metals, and the biota. This investigation focused on the organic exudates of the toxic cyanobacteria Cylindrospermopsis raciborskii as a Cu carrier through a three-level aquatic trophic chain (bacteria, protozoa, and copepod). DISCUSSION: The effects of bacteria activity and growth on the metal-organic complexes were evaluated through changes in free Cu(2+) ions, total dissolved, and total particulate Cu. To be sure that the added copper would be complexed to the exudates, its complexing properties were previously determined. The cyanobacteria exudate-Cu complexes were furnished to bacteria that were further used as a food source to the protozoan Paramercium caudatum. This was then furnished as food to the copepod Mesocyclops sp. The results showed that, in general, the cyanobacterial exudates decreased Cu bioavailability and toxicity to the first trophic level (bacteria), but because the heterotrophic bacteria accumulated Cu, they were responsible for the transference for the otherwise low availability metal form. Both the bacteria and protozoan organisms accumulated Cu, but no metal accumulation was detected in the copepods.

Effects of dietary copper on life-history traits of a tropical freshwater cladoceran.

Life-history parameters of Ceriodaphnia cornuta (Cladocera: Daphniidae) fed on Pseudokirchneriella subcapitata (Chlorophyceae) exposed to different copper concentrations were investigated. C. cornuta individuals were reared in four treatments: (a) reconstituted water and non-contaminated algae (RW); (b) reconstituted water and copper-contaminated algae with either 1.28 × 10(-13) (10(-7)Cu) or (c) 1.93 × 10(-13) g Cu cell(-1) (10(-6)Cu); and (d) natural water from a local reservoir and non-contaminated algae (NW). Copper content in C. cornuta individuals increased as diet-borne exposure increased (RW < 10(-7)Cu < NW < 10(-6)Cu), except for NW individuals, which exhibited higher copper body burden than RW and 10(-7)Cu individuals, suggesting that some copper was available in the natural water. The results suggest that subacute levels of dietary copper stimulated C. cornuta's growth and reproduction, whereas organisms reared on reconstituted water showed nutritional deficiency. Depending on copper exposure concentration, either growth (lower Cu concentration) or reproduction (higher Cu concentration) was further stimulated, suggesting that an alteration of resource allocation is involved in diet-borne copper exposure. Because differences among treatments were only significantly different after day 12 of the experiment, our results reinforce that full life-cycle tests are more appropriate than the standard 7 day or three-brood chronic bioassays used to evaluate dietary copper effects at low, chronic copper inputs and that the use of standard test-organisms may not address site-specific situations for tropical environments.

The effects of Anabaena spiroides exopolysaccharides on copper accumulation in an aquatic food chain.

The influence of Anabaena spiroides exopolysaccharides (EPS) on copper speciation (total dissolved, particulate and free Cu(2+) ions) and bioavailability in aquatic organisms was investigated. Bacteria were used as the first trophic level, Paramecium caudatum (protozoan) as the second and the copepod cyclopoid Metacyclops mendocinus as the third level. The organisms were obtained from a freshwater reservoir and held under continuous laboratory controlled conditions. Freshwater media containing EPS excreted by A. spiroides (10mgL(-1)) and copper (1.0x10(-6)molL(-1)) were used for bacteria growth. Contamined bacteria were used as food source to protozoan, which was further furnished to copepods. The results showed a reduction of EPS concentration during bacteria growth and also a smaller copper accumulation by microorganisms in the presence of EPS. We concluded that A. spiroides exopolysaccharides have reduced copper entrance into the experimental aquatic microbial food chain.

Natural DOM affects copper speciation and bioavailability to bacteria and ciliate.

This study aimed to investigate the influence of natural dissolved organic materials (DOM) on copper speciation (total dissolved, particulate, and free Cu2+ ions) and bioavailability during a two-level experimental microbial food chain. Bacteria were used as the first trophic level, and Paramecium caudatum (protozoan) as the second. The organisms were obtained from a freshwater reservoir and kept under controlled laboratory conditions. Three experimental treatments were performed: exposure of the organisms to copper in the absence of DOM, exposure to DOM in the absence of copper, and exposure to both copper and DOM. Freshwater medium containing natural DOM and copper at a total dissolved concentration of 1.8 x 10(-6) mol L(-1) was furnished to bacteria, which was further used as food to the protozoan. The results showed that after bacterial growth, DOM concentration decreased as quantified by total organic carbon determinations. At the same time, free Cu2+ ions concentration increased in the medium. A lower copper concentration was detected in both microorganisms in the presence of DOM. We conclude that natural DOM reduced copper accumulation in the organisms on the first and second trophic levels, thus reducing the entrance of copper into the aquatic microbial food chain.

The effects of humic substances on copper toxicity to Ceriodaphnia silvestrii Daday (Crustacea, Cladocera).

A major question in the field of ecotoxicology is how DOM affects copper accumulation and toxicity in planktonic organisms; copper acute toxicity and bioaccumulation in Ceriodaphnia silvestrii were investigated in the presence and absence of humic substances (HS) under controlled laboratory conditions. Copper was determined as free Cu2+ ions in the media and total copper in the animals; metal ion buffers were used for ion selective electrode calibration, extending the lower detection limit to 10(-11) mol l(-1). Groups of 20 adult females of similar sizes were exposed (24 h) to a range of nominal copper concentrations. Based on total added copper, LC50 was 4.4 x 10(-8) mol l(-1) without HS, whereas with 20 mg l(-1) HS, it was 25 times higher (1.1 x 10(-6) mol l(-1)). Based on free Cu2+ ions LC50 was statistically similar either with (2.8 x 10(-8) mol l(-1)) or without HS (3.3 x 10(-8 )mol l(-1)). The present results showed that natural DOM reduced copper toxicity and that free Cu2+ ions correlates to the bioavailable fraction to zooplankton. Nevertheless, copper bioaccumulation by C. silvestrii was similar either in the presence or absence of humic substances, suggesting that C. silvestrii regulates its body copper content up to 3.0 x 10(-8) mol l(-1) free Cu2+ ions in the media. The organisms were not able to deal with higher free Cu2+ ions concentrations in the media.

Effects of the herbicide bentazon on growth and photosystem II maximum quantum yield of the marine diatom Skeletonema costatum.

Chlorophyll fluorescence techniques are used for the detection of toxic substances in samples of photosynthetic cells by measuring chlorophyll a fluorimetric parameters, which are a response of the PSII physiological status. This work was conducted to determine the effects of the herbicide bentazon (CASRN 25057-89-0) on growth and maximum quantum yield of photosystem II (Fv/Fm) in cells of the marine diatom Skeletonema costatum. Unialgal cultures were exposed to several bentazon concentrations and its effects on algal growth and Fv/Fm were determined. The traditional algal growth inhibition test (algal biomass measurements) and DCMU-induced chlorophyll a variable fluorescence measurements were determined. Our results showed that even low concentrations of bentazon rapidly lead to Fv/Fm decrease, while the effects on algal growth were detected after 24 h of exposure. The LOEC (2.81 mg L(-1)) and EC50 (13.0 mg L(-1)) determined through Fv/Fm experiments were lower than the LOEC (22.5 mg L(-1)) and EC50 (24.0 mg L(-1)) determined through algal growth inhibition experiments. This confirms that the Fv/Fm is a more sensitive parameter than algal growth for monitoring the effects of bentazon. The present results have demonstrated the applicability of Fv/Fm parameter to access the early toxicity of bentazon, as well as other PSII-inhibition compounds, before significant changes occurred in the original concentration and bioavailability of these toxicants during longer exposure times.

Copper and cadmium complexation by high molecular weight materials of dominant microalgae and of water from a eutrophic reservoir.

High molecular weight materials (HMWM, >12000-14000 Da) excreted by the two cyanophyte species (Microcystis aeruginosa and Anabaena spiroides) and a diatom (Aulacoseira granulata) which are dominant phytoplankton species in a eutrophic reservoir, Barra Bonita, Brazil were investigated as copper (Cu) and cadmium (Cd) complexation agents and their monosaccharide and elemental analysis of C, H, N and S determined. Also, HMWM obtained from the reservoir water as well as from a mixture of the three algae materials were studied. The HMWM of the cyanophytes and the mixture of the three algae materials complexed Cu and Cd, whereas the HMWM of the diatom and that from the reservoir water complexed only Cu. Two classes of ligands of intermediate to weak binding strength were obtained after Scatchard plot analysis of the titration data. The cyanophytes and the mixture HMWM presented higher conditional stability constants for Cu class-1 ligands (logK1' = 9.2-9.5) than the HMWM derived from the diatom and the reservoir water (logK1' = 8.6-8.8). Higher proportions of acidic monosaccharides corresponded to higher K1' of Cu and Cd complexation, yet no relation was observed among complexation parameters and elemental analysis. This study points out Cu ligands of intermediate to weak binding strength in the excreted HMWM of dominant microalgae and in the HMWM of the reservoir water, while Cd was solely complexed by ligands isolated from the cyanophyte HMWM.

Copper complexing properties of dissolved organic materials exuded by the freshwater microalgae Scenedesmus acuminatus (Chlorophyceae).

Dissolved organic materials released by the freshwater microalgae Scenedesmus acuminatus were fractionated into low- and high-molecular weight materials, which were investigated for their capacity to bind copper. The high-molecular weight material was also investigated for its monosaccharide composition and is further discussed in relation to the copper binding property. S. acuminatus was grown in batch cultures under laboratory controlled conditions and harvested at the beginning of stationary growth phase when exuded organic materials were obtained. Copper-complexing property of the total exuded organic materials and exopolysaccharides before and after freeze-drying was evaluated by complexometric titrations and Scatchard Plot Analysis of the titration data. The results revealed the presence of two copper-complexing ligands in the total exuded material, but only one in the exopolysaccharide. Stronger copper-complexing ligands are associated to low molecular weight compounds (LogK'1=7.3, LogCL1=-5.6; LogK'2=6.3, LogCL2=-5.1), whereas weaker ligands to the high molecular weight fraction (LogK'2=6.4, LogCL2=-5.6). Although freeze-drying the polymeric organic material (exopolysaccharide) may result in conformational changes of the molecule, no effect on copper-complexing properties was detected. Gas chromatography was used to evaluate the monosaccharide composition of the microalgal exopolysaccharide, which detected high content of mannose and 12% of acid monosaccharides.

Bioleaching of metals from anaerobic sewage sludge: effects of total solids, leaching microorganisms, and energy source.

The effects of municipal sewage sludge solids concentration, leaching microorganisms (Thiobacillus thiooxidans or Thiobacillus ferrooxidans) and the addition of energy source (S0 or Fe(II)) on the bioleaching of metals from sewage sludge has been investigated under laboratory conditions using shake flasks. The results show that metal solubilization was better accomplished if additional energy source is supplemented to the microorganisms and that T. thiooxidans furnishes, in general, more adequate conditions for the bioleaching than T. ferrooxidans. At a total solids concentration of 70 g L-1 (originally present in the sludge) pH drop and ORP increase are attenuated, so metal solubilization is negatively affected. It was also demonstrated that if lead (Pb) solubilization is to be achieved, than a special combination of microorganism/energy source must be applied.

An evaluation into the potential of biological processing for the removal of metals from sewage sludges.

The use of sewage sludge in agricultural land as a means of sludge disposal and recycling has been shown to be economical and suitable because of the presence of nutrients such as nitrogen and phosphorus. However, municipal sludges often contain high quantities of toxic metals and other compounds that must be removed for its safe use in agricultural soils. The biological leaching of metals from sewage sludges has been shown to be a promising technique for metal detoxifying in such complex matrix. The process efficiency is dependent on several physico-chemical parameters, such as total solids concentration, metal forms, pH-ORP, and temperature. Scale-up of the process has not yet been defined and is still pursuing the correct operational design. Current research involving the bioleaching of metals from sewage sludge and its application to land, which affects soil physical properties, are presented and discussed.