Dynamic of CO2, CH4 and N2O in the Guadalquivir estuary.

The concentration of dissolved CH4 and N2O, as well as the partial pressure of CO2 (pCO2) were studied in the Guadalquivir estuary. Samples were taken in March and April 2018 and 2019, under different rainy and tidal conditions. The available database for summer 2017 (Sierra et al., 2020) was included in the interpretation of the factors that determine the variability of these gases in the Guadalquivir estuary. Two different types of samplings were carried out: a longitudinal transect across the river with salinity values close to zero and another one during two consecutive tidal cycles in the mouth of the estuary. The highest concentrations were found in the upper zone of the estuary and during the low tide. This distribution was related to 4 factors: temperature, salinity, exchange with the atmosphere, and biochemical processes together with the river inputs. Temperature is one of the factors that clearly seems to determine the distribution of gases and fluxes, showing the highest values in the upper zone during the summer of 2017. Intense rains cause a dilution effect of the gas in the water column, this provoked, during the season of spring 2018, an increase in the salinity factor in the distribution of gases in the middle zone. High concentrations of the gases have been linked to production processes in the water column, as well as to benthic production and lateral inputs. While the gases concentrations at the mouth presented values close to those of the equilibrium with the atmosphere, the fluxes in the upper zone of the estuary reached average values of 89.6 mmol m-2 d-1, 121.7 µmol m-2 d-1 and 59.9 µmol m-2 d-1 for CO2, CH4 and N2O, respectively. Generally, water-atmosphere fluxes are positive through the whole study, which means that the estuary acts as a source of these gasses to the atmosphere.

"Greenhouse gas dynamics in a coastal lagoon during the recovery of the macrophyte meadow (Mar Menor, SE Spain)".

The Mar Menor is a hypersaline coastal lagoon with salinity values ranging from 41.9 to 45.5. The system is subjected to a high anthropic pressure that causes an intense eutrophication process, followed by a recovery of the macrophyte meadows. This study focuses on the distribution of the main greenhouse gases (CO2, CH4 and N2O) and was carried out in the extreme seasonal conditions of winter and summer during the year 2018. Sediment-water-atmosphere exchanges and biochemical processes in the water column appeared to be the main factors to explain the variability of these gases. Dissolved Inorganic Carbon (DIC), CH4 and N2O benthic fluxes values obtained in this study, were of 91 ± 29 mmol m-2 d-1, 3.9 ± 1.9 µmol m-2 d-1 and -0.65 µmol m-2 d-1, respectively, along with an important seasonal variation observed, with an increase of DIC and CH4 benthic fluxes during the summer season. Mean values of partial pressure of CO2 (pCO2) in surface water were of 579 µatm in winter and 464 µatm in summer, therefore we can establish that the Mar Menor acts as a source of this gas emitting 3.3 ± 3.0 mmol CO2 m-2 d-1 to the atmosphere. In spite of this, the Mar Menor has a strong autotrophic behaviour partly due to the recovery of the macrophyte meadows, presenting an estimated NEP of 101 mmol m-2 d-1. Regarding to CH4, the mean fluxes to the atmosphere were of 8.0 ± 5.8 µmol m-2 d-1 and there was evidence of CH4 production in the water column that increased in summer. Last of all, in the case of N2O the system acts as a sink with values of -0.65 ± 0.5 µmol m-2 d-1, presenting an intake of N2O that is usually detected in pristine systems.

Distribution of N2O in the eastern shelf of the Gulf of Cadiz (SW Iberian Peninsula).

Distribution of N2O has been determined in eight cruises along three transects (Guadalquivir, Sancti Petri and Trafalgar) in the Gulf of Cadiz, during 2014 and 2015. The mean N2O value for this area was 10.0±0.9nM, with large spatial and temporal variations. Stratification in the water column has been observed; the concentration of this gas increases with the depth, because of the presence of the Eastern North Atlantic Central Water (ENACW) and the Mediterranean Outflow Waters (MOW). The N2O production measured in this study is mainly due to nitrification. N2O yields from nitrification were estimated from the linear correlation of the excess of N2O (ΔN2O) with Apparent Oxygen Utilization (AOU) and nitrate (NO3-), with values of their slopes ranged between 0.010 and 0.021% and 0.017-0.025% respectively. There is an onshore - offshore gradient of N2O; the highest values were found at the shallower stations, indicating coastal input and benthic remineralization. The seawater-air flux of N2O is affected by several variables (temperature, AOU and NO3-), and the average flux calculated is 2.7±2.0µmolm-2d-1. The fluxes show a decrease with increasing distance from the coast, and with proximity to the Strait of Gibraltar. The study area behaves as a source of N2O to the atmosphere, with a global emission of 0.18Ggyear-1.

Spatial and seasonal variability of CH4 in the eastern Gulf of Cadiz (SW Iberian Peninsula).

Methane (CH4) concentrations were measured along three sections of the eastern Gulf of Cadiz (designated "Guadalquivir", "Sancti Petri" and "Trafalgar") during eight cruises in 2014 and 2015. The concentration of CH4 varied from 3.6 to 19.7nmolkg-1 (CH4 saturation percent of 122 and 916%), showing seasonal variation. The highest values were found in December 2014 and November 2015. In most of the sampling area the highest concentration of CH4 was found in subsurface waters at depths close to the thermocline, and in the bottom waters near the coast. The seawater-air flux of CH4 ranged between 0.8 and 59.7µmolm-2d-1, showing seasonal variation in function of the temperature of the surface water. In the "Guadalquivir" and "Sancti Petri" sections, the CH4 fluxes increased with proximity to the coast; this may be a result of continental inputs and CH4 emissions from sediments. The whole study area behaves as a source of CH4 to the atmosphere with mean values of 0.5 and 0.6GgCH4yr-1 in 2014 and 2015, respectively.

Aerobic biodegradation of linear alkylbenzene sulfonates and sulfophenylcarboxylic acids for different salinity values by means of continuous assays.

Aerobic biodegradation of linear alkylbenzene sulfonates (LAS) and sulfophenylcarboxylic acids (SPCs) in water, at different salinity values, has been studied. Three experiments have been carried out employing a staircase model system with continuous dosage of LAS to the system and using concentrations of LAS of the same order as those detected in littoral waters receiving urban wastewater discharges. LAS biodegradation was observed to be almost complete (showing a great extent), and in all cases exceeds 98.4%. At the very low concentration values of LAS utilized in the experiments, no significant variations in the biodegradation of LAS due to the effect of the different salinity values assayed were observed. The biodegradation intermediates detected for all the cases were sulfophenylcarboxylic acids with carboxylic chains of between five and 13 carbon atoms. The detection of C13-SPC (which is only produced by C13-LAS) confirms the existence of omega-oxidation. The total disappearance of SPCs in all cases indicates that mineralization of LAS at the concentrations tested was complete.

Oxidative stress and histopathology damage related to the metabolism of dodecylbenzene sulfonate in Senegalese sole.

Surfactants such as linear alkylbenzene sulfonates (LAS) are widely utilised in the formulation of detergents in commercial products. After use, they pass through waste water treatment plants (WWTP) and are then discharged to aquatic ecosystems, causing risk to aquatic life. The exposure of marine animals to these compounds enhances the production of reactive oxygen species (ROS) with subsequent damage to macromolecules, and produces histological alterations. A flow-through experiment with Senegalese sole (Solea senegalensis) has been devised with the object of correlating the metabolism of LAS including sulfophenylcarboxylic acids (SPCs) by fish with their antioxidant defence system (generation of oxyradicals) and histopathological damage. The generation of intermediate degradation products (SPCs) by the organism, the histopathological responses, the antioxidant enzymes (catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GR), and glutathione S-transferase (GST)), as well as other kinds of enzyme such as acid and alkaline phosphatases (AcP, ALP), were measured. SPCs from 5ØC(6) to 11ØC(12) were identified and quantified in fish and water; their concentrations differed depending on the sampling moment. In general, the responses found in the enzymes were slight: a decrease in the enzymatic activity in gills and activation in the digestive tract. The evidence of histopathological damage identified was also small; the organism's defensive mechanism against pollutants should enable it to recover easily. A direct relationship was established between biotransformation and the generation of SPCs and ROS. In conclusion, the correct functioning of the antioxidant defence system with absence of large variations, the short-term histopathological damage, and the evidence of SPCs indicate an adequate metabolism of 2-phenyl-C(12)-linear alkylbenzene sulfonates (2ØC(12)LAS) by this specie and non-toxic effects at environmentally realistic levels.

Testing organic solvents for the extraction from fish of sulfophenylcarboxylic acids, prior to determination by liquid chromatography-mass spectrometry.

The present paper describes the use of different solvent mixtures to extract from fish various sulfophenylcarboxylic acids (SPCs of C(6) to C(13)), and their originating compounds, linear alkylbenzene sulfonates (LAS of C(10) to C(13)). The analytical method utilized involves pressurized liquid extraction, followed by preconcentration of the samples, purification by solid-phase extraction, and finally identification and quantification of the target compounds by high-performance liquid chromatography-mass spectrometry using a system equipped with an electrospray interface operating in negative ion mode. The SPCs and LAS were extracted from spiked fish first with hexane to remove interference from fats, then with different mixtures of solvents: dichloromethane followed by methanol; 50:50 dichloromethane-methanol; and 30:70 dichloromethane-methanol. The LAS recoveries obtained with these three extraction options were high (between 68.5 and 80.8%); however, owing to the low percentages obtained for SPC homologues (13.5, 13.1, and 15.9%, respectively), another extraction procedure with methanol was developed in order to increase these recoveries. The percentage of recovery for total SPCs with the methanolic extraction was higher (90.1%), with a standard deviation of 9.9, and the LAS recoveries also increased (99.9%). Detection limits were between 1 and 22 ng g(-1) for LAS, and between 1 and 58 ng g(-1) for SPCs. Quantitation limits were between 4 and 73 ng g(-1) for LAS, and between 2 and 193 ng g(-1) for SPCs. This method has been applied to measure the biotransformation of 2ØC(10) LAS (where Ø is a sulfophenyl group) in fish exposed in a flow-through system, and enabled the separation and identification of SPCs from 5ØC(6) to 9ØC(10).

Presence, biotransformation and effects of sulfophenylcarboxylic acids in the benthic fish Solea senegalensis.

The presence of linear alkylbenzene sulfonates (LAS) and their degradation intermediates, sulfophenylcarboxylic acids (SPCs), with concentrations up to 100 ppb has been found in surface waters taken from the estuary of the river Guadalete (Cádiz, SW of Spain). Higher concentrations were found at the sampling site located adjacent to the discharge outlet of a wastewater treatment plant (WWTP). The concentrations decreased downstream to a few ppb as a result of dilution, sorption, and degradation processes, which were taking place along the estuary. Once the presence of both xenobiotics was confirmed in the environment, an in vivo assay was conducted to study their biotransformation and effects in the benthic fish Solea senegalensis. A flow-through system was employed, consisting of an exposure phase (120 h) with environmental levels of the surfactant (200, 500 and 800 microg/L of 2ØC(10)LAS), followed by a depuration stage (72 h). The generation of SPCs has been quantified during these phases in both water and fish, with LAS biotransformation shown in all cases. The antioxidant enzymes catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GR), the phase II enzyme glutathione S-transferase (GST), and the phase III acid and alkaline phosphatases (AcP, ALP) were also estimated and utilized as biomarkers.

Presence of surfactants and their degradation intermediates in sediment cores and grabs from the Cadiz Bay area.

The occurrence and distribution of the major surfactants--LAS, AES, APEOs and AEOs--and their degradation intermediates--SPCs, AP and APECs--in a marine-estuarine environment at Spain are presented. Results show that their concentration in surface sediments is clearly correlated with their usage and the existence of wastewater discharges. The degradation processes appear to lead to the formation of SPCs in the case of LAS, and to the shortening of the average ethoxylated chain length in the case of NPEOs, AEOs and AES. Vertical profiles for AEOs and AES are reported for the first time and present the highest values nearest the surface, followed by a sharp decrease with depth for all surfactants, as well as the appearance of degradation intermediates in deeper sedimentary layers. Shorter LAS homologues and SPCs tend to be present in pore water while strongly non-polar intermediates like NP are firmly attached to the sediments.

New extraction method for the analysis of linear alkylbenzene sulfonates in marine organisms. Pressurized liquid extraction versus Soxhlet extraction.

A new method has been developed for the determination of linear alkylbenzene sulfonates (LAS) from various marine organisms, and compared with Soxhlet extraction. The technique applied includes the use of pressurized liquid extraction (PLE) for the extraction stage, preconcentration of the samples, purification by solid-phase extraction (SPE) and analysis by liquid chromatography with fluorescence detection. The spiked concentrations were added to the samples (wet mass of the organisms: Solea senegalensis and Ruditapes semidecussatus), which were homogenized and agitated continuously for 25 h. The samples were extracted by pressurized hot solvent extraction using two different extraction temperatures (100 and 150 degrees C) and by traditional Soxhlet extraction. The best recoveries were obtained employing pressurized hot solvent extraction at 100 degrees C and varied in the range from 66.1 to 101.3% with a standard deviation of between 2 and 13. Detection limit was between 5 and 15 microg kg(-1) wet mass using HPLC-fluorescence detection. The analytical method developed in this paper has been applied for LAS determination in samples from a Flow-through exposure system with the objective of measuring the bioconcentration of this surfactant.

Simulating a heavy metal spill under estuarine conditions: effects on the clam Scrobicularia plana.

We describe the effect of heavy metals Zn, Cd, Pb and Cu on the induction of methallothioneins on the clam Scrobicularia plana along a salinity gradient simulated under laboratory conditions. The clams were exposed to constant heavy metal concentrations in a dynamic estuary simulator during a 15-day assay to investigate possible induction of metal-binding proteins in them. The concentration of heavy metals in water was analysed. Clams were analysed for methallothionein concentrations. The speciation of Zn, Cd, Pb and Cu along the salinity gradient was modelled. Zn showed the highest concentrations and its prevalent species was the free ion. Intersite differences have been observed in methallothionein concentration and related to the salinity gradient. It seems that synthesis of methallothioneins is the result of physiological forces acting in concert with the changes in the chemical speciation of metals, owing to the trace metals uptake is controlled by means of an interaction of physiology and physicochemistry.

The behaviour of heavy metals from the Guadalquivir estuary after the Aznalcóllar mining spill: field and laboratory surveys.

The spill at 'Los Frailes' caused a large input of metals into the riverine, estuarine and coastal ecosystems. Metal behaviour in natural waters can be studied both by performing field measurements, and by undertaking laboratory simulation experiments. Here is described the behaviour of the heavy metals Zn, Cd, Pb and Cu in the Guadalquivir estuary using incubation experiments and the results compared to those obtained during field assessments. If the toxic waste water had been discharged untreated into the environment, it could have raised the heavy metal concentrations in the estuary water to high values, such as 20.65 microM for Zn and 40 x 10(-3) microM for Cd. These values are several times higher than those obtained in samples collected in the area during the monitoring of the early impact. The metals present a non-conservative behaviour, with a high loss to waters of low salinity values.

Evaluating the heavy metal contamination in sediments from the guadalquivir estuary after the Aznalcóllar mining spill (SW Spain): a multivariate analysis approach.

The Doñana National Park is one of the most important Natural Reserves in west Europe. The park and the Guadalquivir estuary were impacted by the release of 5 million cubic meters of acid waste from the processing of pyrite ore. Here are presented a multivariate analysis approach to evaluate the extension and the magnitude of the accidental spill on the estuarine sediments. Sediments were used to analyze 6 different metals (Fe, Mn, Zn, Cd, Pb, Cu). Also the concentration of the metals in the different geochemical sediment fractions were determined as a means of assessing the potential impact of the acid water from the mine. The analysis of the results obtained permits to establish that the impact of the accident was acute on the ecosystem due to the set up of a treatment plan in June 1998 and to the high hydrodynamic of the estuary that absorb the early impact. Chronic contamination by Cu was also detected during the monitoring of the impact of the accident in the estuary but this did not appear to be related to the accidental spill. Because the likelihood of long term adverse effects from the heavy metal contamination of estuarine sediments, further ecotoxicological evaluations should be carried out for the estuary and the park using an integrated approach.

Occurrence and distribution of linear alkylbenzene sulfonates and sulfophenylcarboxylic acids in several Iberian littoral ecosystems.

The distribution of linear alkylbenzene sulfonates (LAS) and sulfophenylcarboxylic acids (SPC) has been characterized in water and sediment of five Iberian estuaries and a saltmarsh channel. The highest concentrations of LAS and SPC were detected in shallow zones close to discharge points of untreated urban effluent, and decreased sharply with increasing distance from these. SPC homologues of between 3 and 12 carbon atoms were detected, although their concentrations were several orders of magnitude lower than the corresponding LAS. The highest SPC concentrations found were of medium carboxylic chain lengths (C6-C8) in water, and of longer chain lengths (C9-C11) in particulate material. LAS homologues were associated preferentially with the particulate material. The vertical distribution of LAS in the water column was not homogeneous, in contrast to that presented by the SPC homologues. In the case of the Sancti Petri Channel, there was a net flow of LAS exportation into the Bay of Cádiz, of -38.6 kg day(-1), and a net flow for SPC of -6.0 kg day(-1). The daily mass input of LAS and their biodegradation intermediates from the Sancti Petri Channel to Cadiz Bay was 44.6 kg.

Influence of the Aznalcóllar mining spill on the vertical distribution of heavy metals in sediments from the Guadalquivir estuary (SW Spain).

The Natural Park of Doñana and the Guadalquivir estuary were impacted by the release of 6 million cubic meters of acid waste after the mine-tailing spill in Aznalcóllar (Andalusia, SW, Spain). Here is presented the monitoring of the accidental spill on vertical distribution of heavy metals in the estuarine sediments. The total concentration of six metals (Fe, Mn, Zn, Cd, Pb, Cu), their chemical speciation and the organic carbon concentration were analyzed in sediment vertical profiles. The results obtained determine background levels similar to previously reported in the area. The analysis catalogues the impact of the accident on the estuary as acute and mainly associated with high concentrations of Zn and Cd. The recent enrichment in Zn and Cd and their geochemical association with the more mobile fractions of the sediment determine an environmental risk associated with the acute impact and detected in some of the areas of the estuary.

Determination of sulfophenylcarboxylic acids in marine samples by solid-phase extraction then high-performance liquid chromatography.

An analytical method is presented for the determination of sulfophenylcarboxylic acids (SPC) produced by the biodegradation of linear alkylbenzene sulfonates (LAS) in marine samples. Isolation and concentration of the compounds was by solid-phase extraction. The different factors affecting extraction efficiency packing composition, pH, clean-up, ionic strength, and elution solvents--were studied and optimized. With the proposed method C4-C13SPC and C10-C13 LAS recoveries varied between 65% and 105%, with standard deviations between 0.1 and 5, respectively, for 100-mL samples and 100 microg L(-1) concentrations of each homolog. Detection limits within the range 0.5 g L(-1) (for C4SPC) to 1.0 g L(-1) (for C12SPC) were obtained by liquid chromatography with fluorescence detection. This method is the first to be proposed that enables the simultaneous determination of monocarboxylic SPC (C>3) and LAS homologs in marine samples by a simple, sensitive, and specific method giving high recoveries and reproducibility. SPC with from three to twelve carbon atoms in the carboxyl chain have been found in marine water samples.

Bioconcentration of linear alkylbenzene sulfonates and their degradation intermediates in marine algae.

Sorption experiments using different homologues of linear alkylbenzene sulfonate (LAS) and sulfophenylcarboxylic acid (SPC) on several marine microalgae have been carried out. The steady state seems to be reached in the first 4 hours. Longer exposure times lead to biodegradation of the compound and, therefore, to an overestimation of the bioconcentration factor. Sorption coefficients are higher for Nannochloropsis gaditana, for example, 1,293 Lkg(-1) for C11-LAS and 525 Lkg(-1) for C11-SPC versus 727 Lkg(-1) for C11-LAS and 28 Lkg(-1) for C11-SPC for Dunaliella salina. For both algae an increase in the sorption coefficient is observed when the polarity of the compound decreases (C5-SPC

Extraction and isolation of linear alkylbenzene sulfonates and their intermediate metabolites from various marine organisms.

A method has been developed for the determination of linear alkylbenzene sulfonates and its degradation intermediates (sulfophenylcarboxylic acids) from different marine organisms, which includes the stages of extraction and analysis by liquid chromatography with fluorescence detection. The extraction stage (Soxhlet and solid-phase extractions) was optimised by the selection of the appropriate solvent, minicolumns and different clean-up stages. Recoveries varied in the range from 80 to 104%, with a standard deviation between 1 and 9%. Detection limits were 15 ng g(-1) wet mass for undecylbezene sulfonate and 30 ng g(-1) wet mass for sulfophenylundecanoic acid using HPLC-fluorescence detection. The complete analytical method was successfully applied to different marine organisms from the Bay of Cadiz (SW Spain).