Long-term evaluation of potential Al toxicity after an Al treatment in a coastal bay.

Coastal areas often suffer from eutrophication, causing ecosystem degradation and oxygen deficiencies. In hundreds of lakes, aluminium (Al) treatment has been a successful method to bind phosphorous in the sediments, reducing lake productivity. In this study we follow up a successful Al treatment of the sediment of Björnöfjärden, which was the first full-scale coastal remediation project using a geo-engineering method, that substantially reduced P concentrations in the water column. We evaluate the long-term development of Al in the water and aquatic life using 10 years data from before, during and after the aluminium treatment. Still after ten years, the treatment is successful with low P concentrations in the Bay. After a temporal increase of Al in water and biota (fish and algae) in connection with the Al treatment, the concentration decreased rapidly to pre-treatment levels. A risk assessment for biota and humans consuming fish and water from the bay showed that the risk for negative effects were negligible, also during the treatment year.

Managing multi-functional peri-urban landscapes: Impacts of horse-keeping on water quality.

Eutrophication assessments in water management to quantify nutrient loads and identify mitigating measures seldom include the contribution from horse facilities. This may be due to lack of appropriate methods, limited resources, or the belief that the impact from horses is insignificant. However, the recreational horse sector is growing, predominantly in multi-functional peri-urban landscapes. We applied an ecosystem management approach to quantify nutrient loads from horse facilities in the Stockholm Region, Sweden. We found that horses increased the total loads with 30-40% P and 20-45% N, with average area-specific loads of 1.2 kg P and 7.6 kg N ha-1 year-1. Identified local risk factors included manure management practices, trampling severity, soil condition and closeness to water. Comparisons of assessment methods showed that literature standard values of area-specific loads and water runoff may be sufficient at the catchment level, but in small and more complex catchments, measurements and local knowledge are needed.

Fish community responses to restoration of a eutrophic coastal bay.

Interest in coastal restoration measures is increasing, but information about subsequent ecosystem recovery processes is limited. In Björnöfjärden on the Baltic Sea coast, Stockholm archipelago, a pioneering case study to reduce coastal eutrophication led to improvements and initially halved phosphorus levels. Here, we evaluate the effects of the restoration on the local fish assemblage over one decade after the measures. The study gives a unique possibility to evaluate responses of coastal fish to nutrient variables and abatement in a controlled natural setting. Cyprinid abundance decreased and perch partially increased with decreasing turbidity levels, while mean trophic level increased over time in the restored area. Responses were overall weak, likely attributed to an attenuation of the eutrophication abatement effect over time. The results suggest that nutrient reduction gives slow responses in fish compared to alternative measures such as fishing closures.

Remediation of a Eutrophic Bay in the Baltic Sea.

Eutrophication of coastal ecosystems is a global problem that often results in bottom water oxygen deficiency and in turn promotes sediment phosphorus (P) release (A). In order to increase sediment P retention, we injected dissolved aluminum into the anoxic sediment of a eutrophic semienclosed bay in the Baltic Sea, thereby inhibiting P recycling and further eutrophication (B). The P concentration in the bay remained at half, as did phytoplankton biomass (C), compared to pretreatment conditions and compared to the reference bay. Four years after treatment the water column transparency was increased, allowing submerged vegetation to penetrate deeper, and the habitat suitable for fish and benthic fauna had expanded (D). The lowered P concentration in the bay decreased the P export to the surrounding archipelago. This is the first full-scale marine remediation project using a geo-engineering method that demonstrates a quick recovery. For successful remediation in coastal areas, permanent binding of mobile P in anoxic sediments may be needed together with measures in the catchment area to obtain faster recovery of eutrophicated marine ecosystems.

Separation of inositol phosphate isomers in environmental samples by ion-exchange chromatography coupled with electrospray ionization tandem mass spectrometry.

A method for isomeric separation of inositol phosphates (InsPn) in environmental samples originating from different sources such as soil, manure/compost, and aquatic sediments has been developed. The method includes a single NaOH-EDTA extraction step, centrifugation and direct injection of a particle free solution into an ion chromatographic column. Isomeric liquid chromatographic separation was achieved with an ammonium carbonate gradient compatible with electrospray ionization tandem mass spectrometric detection (LC-ESI-MS/MS). The detection limits of the LC-ESI-MS/MS method were between 0.03-0.16µM for the different InsPn, corresponding to 37-99ng P/g sample DW. The method has shown to be able to analyze more than 200 samples from soil, manure and sediment without any severe matrix effects. This will allow future studies of the fate of inositol phosphates in the environment.

Longevity and effectiveness of aluminum addition to reduce sediment phosphorus release and restore lake water quality.

114 lakes treated with aluminum (Al) salts to reduce internal phosphorus (P) loading were analyzed to identify factors driving longevity of post-treatment water quality improvements. Lakes varied greatly in morphology, applied Al dose, and other factors that may have affected overall treatment effectiveness. Treatment longevity based on declines in epilimnetic total P (TP) concentration averaged 11 years for all lakes (range of 0-45 years). When longevity estimates were used for lakes with improved conditions through the end of measurements, average longevity increased to 15 years. Significant differences in treatment longevity between deeper, stratified lakes (mean 21 years) and shallow, polymictic lakes (mean 5.7 years) were detected, indicating factors related to lake morphology are important for treatment success. A decision tree developed using a partition model suggested Al dose, Osgood index (OI, a morphological index), and watershed to lake area ratio (related to hydraulic residence time, WA:LA) were the most important variables determining treatment longevity. Multiple linear regression showed that Al dose, WA:LA, and OI explained 47, 32 and 3% respectively of the variation in treatment longevity. Other variables (too data limited to include in the analysis) also appeared to be of importance, including sediment P content to Al dose ratios and the presence of benthic feeding fish in shallow, polymictic lakes.

Speciation of inositol phosphates in lake sediments by ion-exchange chromatography coupled with mass spectrometry, inductively coupled plasma atomic emission spectroscopy, and 31P NMR spectroscopy.

A method for the detection and speciation of inositol phosphates (InsP(n)) in sediment samples was tested, utilizing oxalate-oxalic acid extraction followed by determination by high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) using electrospray ionization (ESI) in negative mode. The chromatographic separation was carried out using water and ammonium bicarbonate as mobile phase in gradient mode. Data acquisition under MS/MS was attained by multiple reaction monitoring. The technique provided a sensitive and selective detection of InsP(n) in sediment samples. Several forms of InsP(n) in the oxalate-oxalic acid extracted sediment were identified. InsP6 was the dominating form constituting 0.250 mg P/g DW (dry weight); InsP5 and InsP4 constituted 0.045 and 0.014 mg P/g DW, respectively. The detection limit of the LC-ESI-MS/MS method was 0.03 µM InsP(n), which is superior to the currently used method for the identification of InsP(n), (31)P nuclear magnetic resonance spectroscopy ((31)P NMR). Additionally sample handling time was significantly reduced.

Extraction and quantification of phosphorus derived from DNA and lipids in environmental samples.

Understanding the flux and turnover of phosphorus (P) in the environment is important due to the key role P plays in eutrophication and in the ambition to find cost-effective measures to mitigate it. Orthophosphate diesters, including DNA and phospholipids (PLs), represent a potentially degradable P pool that could support future primary production and eutrophication. In this study, extraction techniques were optimized and combined with colorimetric determination of extracted P to provide a selective quantification method for DNA-P and PL-P in agricultural soil, sediment and composted manure. The proposed method is rapid and reproducible with an RSD of <10%. Recovery, evaluated by spiking the sample matrices with DNA and PL standards, was over 95% for both DNA and PLs. The method can be used for the determination of the pool size of the two organic P fractions. Results show that DNA-P comprises 3.0% by weight of the total P (TP) content in the studied soil, 10.4% in the sediment and 8.4% in the compost samples. The values for PL-P are 0.5%, 6.0% and 1.7% for soil, sediment and compost, respectively.

Toxicity of inorganic aluminium at spring snowmelt--in-stream bioassays with brown trout (Salmo trutta L.).

Although the acid load has decreased throughout Scandinavia, acidic soils still mobilise aluminium (Al) that is harmful to brown trout. We hypothesise that there are thresholds for Al toxicity and that the toxicity can be traced from the water content to gill accumulation and the consequential physiological effects. During snowmelt, yearlings were exposed to a gradient of pH and inorganic monomeric Al (Al(i)) in humic streams to study the toxic effects and mortality. Gill Al and physiological blood analyses [haemoglobin (Hb), plasma chloride (P-Cl) and glucose (Glu)] were measured. As the water quality deteriorated, Al accumulated on the gills; Hb and Glu increased; P-Cl decreased, and mortality occurred. Moribund fish had significantly increased gill Al and Hb, suggesting that respiratory disturbances contributed to mortality. Decreased P-Cl and plasma availability indicated an ion regulatory disturbance and possibly circulatory collapse. Al(i) should be less than 20 µg/L, and pH higher than 5.0, to sustain healthy brown trout populations. These thresholds can be used to fine-tune lime dose, as both Al(i) and pH levels have to be balanced to prevent harm in the recovering aquatic biota. Although Al is tightly linked to pH, local variation in Al availability in soil and bedrock affects the Al release and subsequent toxic Al(i) episodes in some catchment areas.

Diagenesis of settling seston: identity and transformations of organic phosphorus.

Solution (31)phosphorus NMR spectroscopy and sequential fractionation were used to follow diagenetic changes in phosphorus forms during decomposition of settling seston in Lake Nordborg, a shallow eutrophic lake in Denmark. In a decomposition experiment, seston released >60% of their total phosphorus during ~50 days incubation, although seston collected during summer contained more phosphorus and released it over a longer period compared to seston collected during spring. Seston decomposition increased concentrations of potentially bioavailable polyphosphate and phosphodiesters, but also promoted the formation of refractory phosphorus forms that might be buried permanently in the sediment. Combining these results with in situ measurements of phosphorus concentrations in lake water and sediment traps revealed that the release from settling seston plays only a minor role in the accumulation of phosphorus in the hypolimnion of Lake Nordborg.

Which aluminium fractionation method will give true inorganic monomeric Al results in fresh waters (not including colloidal Al)?

Aluminium solubility and toxicity increase with acidification. There is no standardized analytical method for the determination of inorganic monomeric Al (Al(im)), which is the form that causes toxicity to fish. Separation by cation exchange is commonly combined with other analytical methods, such as complexation with pyrochatechol violet (PCV) or 8-hydroxyquinoline (HQ) and total quantification using graphite furnace or inductively-coupled plasma emission. Data from 14 laboratories were obtained for a dilution series of Al(im) samples; the results of the Al(im) analysis were statistically evaluated. The Al(im) levels were altered through pH, which was controlled by the addition of calcium hydroxide. Confounding parameters such as total organic carbon (TOC) or fluoride (F) were controlled. The total determination and HQ methods yielded significantly higher Al(im) concentrations than the PCV method. Pretreatment by passage through a 0.45 microm filter and pH-adjustment of the ion exchange column had no apparent effect on the Al(im) yield. However, ultra filtration (<10 kDa) caused a significant reduction in the Al(im) concentration using the HQ method. The ultra filtrated Al(im) fraction was similar to the PCV results in the interlaboratory comparison. Retention of colloidal bound Al in the cation exchange column may result in overestimation of Al(im) when the total and HQ methods are used. Estimated Al(im) concentrations derived from two equilibrium models were similar to PCV-derived Al(im) concentrations, as well as the HQ method using ultra filtrated water. The fact that the PCV method does not detect colloidal Al, neither before nor after ion exchange, makes this a preferred technique for Al(im) analysis. Because of the variability in the reported Al(im) concentrations that can arise when different analytical procedures are used, the adoption of a single, reliable technique will facilitate inter-study comparisons and provide consistency in the detection of trends in environmental monitoring programs.

Screening for organic phosphorus compounds in aquatic sediments by liquid chromatography coupled to ICP-AES and ESI-MS/MS.

The structures of organic phosphorous (P) compounds in aquatic sediments are to a large extent unknown although these compounds are considered to play an important role in regulating lake trophic status. To enhance identification of these compounds, a liquid chromatography (LC) method for their separation was developed. The stationary phase was porous graphitic carbon (PGC), and the mobile phases used in the gradient elution were compatible with both inductive coupled plasma atomic emission spectroscopy (ICP-AES) and electrospray ionization tandem mass spectrometry (ESI-MS/MS). With LC-ICP-AES, eight different P containing peaks could be observed in the P chromatogram indicating that at least eight different P compounds were separated. With the setup of an information dependent acquisition (IDA) with ESI-MS/MS, the mass over charge ( m/ z) of compounds containing a phosphate group (H 2PO 3 (-), m/ z 97) could be measured and further fragmentation experiments gave additional information on the structure of almost 40 separated P compounds, several were verified to be nucleotides. ICP-AES was very suitable in the development of the LC method and allowed screening and quantification of P compounds. The presented LC-ESI-MS/MS technique was able to identify several sediment organic P compounds.

Sediment extraction and clean-up for organic phosphorus analysis by electrospray ionization tandem mass spectrometry.

A method to prepare NaOH sediment extracts for organic P compound analysis with electrospray ionization tandem mass spectrometry (ESI-MS-MS) was developed on natural samples. Ion exchange, rotary evaporation and mass cut-off filtering proved to be suitable for sample preparation. Samples were analyzed with ESI-MS-MS, and reproducibility and repeatability of the method was calculated. In addition, (31)P-nuclear magnetic resonance spectroscopy ((31)P NMR) was used to measure recovery of different P compound groups such as orthophosphate (Ortho-P), orthophosphate monoesters (Monoester-P), orthophosphate diesters (Diester-P) and pyrophosphates (Pyro-P). The developed sample preparation method resulted in an easy-to-spray liquid for the ESI-MS-MS instrumentation. The overall P recovery was 65% and (31)P NMR showed that Diester-P, possibly in the form of DNA, was apparently lost through the filtering step most likely due to their size. Variances in the total intensities of the MS scans (relative standard deviation (R.S.D.) 35-54%) were for about 50% due to repeated MS runs. Covariances of the peaks in the MS spectra were calculated to be for about 30% due to the sample preparation procedure. Finally, with the ESI-MS-MS approach, 11 peaks in the mass spectra were found likely to represent phosphate containing compounds.

Sediment phosphorus extractants for phosphorus-31 nuclear magnetic resonance analysis: a quantitative evaluation.

The influence of pre-extractant, extractant, and post-extractant on total extracted amounts of P and organic P compound groups measured with 31P nuclear magnetic resonance (31P-NMR) in lacustrine sediment was examined. The main extractants investigated were sodium hydroxide (NaOH) and sodium hydroxide ethylenediaminetetraacetic acid (NaOH-EDTA) with bicarbonate buffered dithionite (BD) or EDTA as pre-extractants. Post extractions were conducted using either NaOH or NaOH-EDTA, depending on the main extractant. Results showed that the most efficient combination of extractants for total P yield was NaOH with EDTA as pre-extractant, yielding almost 50% more than the second best procedure. The P compound groups varying the most between the different extraction procedures were polyphosphates and pyrophosphates. NaOH with BD as pre-extractant was the most efficient combination for these compound groups.

Biogenic phosphorus in oligotrophic mountain lake sediments: differences in composition measured with NMR spectroscopy.

Phosphorus (P) composition in alkaline sediment extracts from three Swedish oligotrophic mountain lakes was investigated using 31P-NMR spectroscopy. Surface sediments from one natural lake and two mature reservoirs, one of which has received nutrient additions over the last 3 years, were compared with respect to biogenic P composition. The results show significant differences in the occurrence of labile and biogenic P species in the sediments of the different systems. The P compound groups that varied most between these three systems were pyrophosphate and polyphosphates, compound groups known to play an important role in sediment P recycling. The content of these compound groups was lowest in the reservoirs and may indicate a coupling between anthropogenic disturbances (i.e., impoundment) to a water system and the availability of labile P species in the sediment. A statistical study was also conducted to determine the accuracy and reliability of using 31P-NMR spectroscopy for quantification of sediment P forms.

Effects of aluminum treatment on phosphorus, carbon, and nitrogen distribution in lake sediment: a 31P NMR study.

The effects of aluminum (Al) treatment on sediment composition of carbon (C), nitrogen (N) and phosphorus (P) were investigated in sediment representing pre- and post-treatment years in the Danish Lake Sønderby. 31P NMR spectroscopy analysis of EDTA-NaOH extracts revealed six functional P groups. Direct effects of the Al treatment were reflected in the orthophosphate profile revealing increased amounts of Al-P in the sediment layers representing the post-treatment period, as well as changes in organic P groups due to precipitation of phytoplankton and bacteria at the time of Al addition. Furthermore, changes in phytoplankton community structure and lowered production due to the Al treatment resulted in decreased concentrations of sediment organic P groups and total C. Exponential regressions were used to describe the diagenesis of C, N, and P in the sediment. From these regressions, half-life degradation times and C, N, and P burial rates were determined.

Sediment depth attenuation of biogenic phosphorus compounds measured by 31P NMR.

Being a major cause of eutrophication and subsequent loss of water quality, the turnover of phosphorus (P) in lake sediments is in need of deeper understanding. A major part of the flux of P to eutrophic lake sediments is organically bound or of biogenic origin. This P is incorporated in a poorly described mixture of autochthonous and allochthonous sediment and forms the primary storage of P available for recycling to the water column, thus regulating lake trophic status. To identify and quantify biogenic sediment P and assess its lability, we analyzed sediment cores from Lake Erken, Sweden, using traditional P fractionation, and in parallel, NaOH extracts were analyzed using 31P NMR. The surface sediments contain orthophosphates (ortho-P) and pyrophosphates (pyro-P), as well as phosphate mono- and diesters. The first group of compounds to disappear with increased sediment depth is pyrophosphate, followed by a steady decline of the different ester compounds. Estimated half-life times of these compound groups are about 10 yr for pyrophosphate and 2 decades for mono- and diesters. Probably, these compounds will be mineralized to ortho-P and is thus potentially available for recycling to the water column, supporting further growth of phytoplankton. In conclusion, 31P NMR is a useful tool to asses the bioavailability of certain P compound groups, and the combination with traditional fractionation techniques makes quantification possible.