Matrix effect study in the determination of linear alkylbenzene sulfonates in sewage sludge samples

We propose a study of the matrix effect in the determination of linear alkylbenzene sulfonates (LAS) in sewage sludge samples. First, a rapid, selective and sensitive method is proposed. The method involves two stages: the extraction of the compound from the samples and analysis by liquid chromatography with fluorescence detection (LC-FLD). Three different techniques of extraction (microwave-assisted extraction, Soxhlet, and ultrasounds) were compared, and microwave-assisted extraction was selected as the best suited for our purpose. Microwave-assisted extraction allows reducing the extraction time (25?min compared with 12?h for conventional Soxhlet extraction) and solvent waste (25?ml of methanol compared with 200?ml for Soxhlet or more than 50?ml for the ultrasonic procedure). Absence of matrix effect was evaluated with two standards (2?C(8:0) and 2?C(16:0) ) that are not commercial; therefore, neither of them was detected in sewage sludge samples and they showed similar environmental behavior (adsorption and precipitation) to LAS (C(11:0) -C(13.0) ), which allow us to evaluate the matrix effect. Validation was carried out by a recovery assay, and the method was applied to samples from different sources; therefore, they had different compositions.     

Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 5. Probabilistic risk assessment of linear alkylbenzene sulfonates in sludge-amended soils

Linear alkylbenzene sulfonates (LAS) can be found in high concentrations in sewage sludge and, hence, may enter the soil compartment as a result of sludge application. Here, LAS may pose a risk for soil-dwelling organisms. In the present probabilistic risk assessment, statistical extrapolation has been used to assess the risk of LAS to soil ecosystems. By use of a log-normal distribution model, the predicted no-effect concentration (PNEC) was estimated for soil fauna, plants, and a combination of these. Due to the heterogeneous endpoints for microorganisms, including functional as well as structural parameters, the use of sensitivity distributions is not considered to be applicable to this group of organisms, and a direct, expert evaluation of toxicity data was used instead. The soil concentration after sludge application was predicted for a number of scenarios and used as the predicted environmental concentration (PEC) in the risk characterization and calculation of risk quotients (RQ = PEC/PNEC). A LAS concentration of 4.6 mg/kg was used as the current best estimate of PNEC in all RQ calculations. Three levels of LAS contamination (530, 2,600, and 16,100 mg/kg), three half-lives (10, 25, and 40 d), and five different sludge loads (2, 4, 6, 8, and 10 t/ha) were included in the risk scenarios. In Denmark, the initial risk ratio would reach 1.5 in a realistic worst-case consideration. For countries not having similar sludge regulations, the estimated risk ratio may initially be considerably higher. However, even in the most extreme scenarios, the level of LAS is expected to be well beyond the estimated PNEC one year after application. The present risk assessment, therefore, concludes that LAS does not pose a significant risk to fauna, plants, and essential functions of agricultural soils as a result of normal sewage sludge amendment. However, risks have been identified in worst-case scenarios.     

Joint toxicity of linear alkylbenzene sulfonates and pyrene on Folsomia fimetaria

Surfactants may enhance the biodegradation of hydrophobic substances in soils. This has partly been attributed to an increase in the bioavailability, brought about by the presence of surfactants. The aim of this study was to examine the ecotoxicological effects of the detergent linear alkylbenzene sulfonate (LAS) and the polycyclic aromatic hydrocarbon pyrene, alone and in combination, using the survival and reproduction of the collembolan Folsomia fimetaria as endpoints. The EC(50) and EC(10) were 803 and 161 mg kg(-1) for LAS, and 23 and 15 mg kg(-1) for pyrene. If LAS was able to increase the bioavailability of pyrene to springtails, it was expected that the combined effect of the two substances would exceed the effect found for each of the compounds tested separately. However, the results showed no effect of LAS on the toxicity of pyrene in the concentration range tested (1-750 mg LAS kg(-1) dry weight). Both the toxic unit concept and the isobologram method indicated that an additive approach would be the most useful when assessing the risk of these two compounds.     

Effects of linear alkylbenzene sulfonates on functional diversity of microbial communities in soil

Linear alkylbenzene sulfonates (LAS) often occur in sewage sludge that is applied to agricultural soil. Here LAS may affect the microbial activity, which is an important basis for nutrient cycling. Previous studies have shown that single bacterial species and specific soil processes can be very sensitive to LAS. Here we report that two levels of LAS, 22 and 174 mg/kg dry weight soil, had little or no significant influence of the functional diversity of bacteria in a sandy soil, as tested by community-level physiological profiles. Briefly, these profiles are a characterization of the microbial communities based on the pattern of substrate utilization in 96-well microtiter plates (Biolog EcoPlates). Sandy agricultural soil was incubated in duplicate 1-L mesocosms with or without LAS, and bacteria were extracted after one, two, and four weeks. During incubation, more than 98 and 93% of LAS added to 22 and 174 mg/kg dry weight soil was degraded, respectively. The presence of LAS at 174 mg/kg dry weight caused a higher number of bacteria in the soil extracts, maximally corresponding to 2.5 times the numbers in LAS-free soil (1.8 x 10(7) cells/g dry wt soil) after four weeks of incubation. No inhibitory effect of LAS was observed when the substrate utilization data were analyzed for substrate richness and diversity (Shannon-Weaver indices). Principal component analysis, however, showed that the pattern of substrate utilization in soil with the highest LAS content (174 mg/kg dry wt) could be distinguished from control soil and soil with 22 mg LAS/kg dry weight. Yet the overall conclusion was that the functional diversity of the aerobic, heterotrophic bacterial community was rather insensitive to LAS.     

Uptake, distribution, and elimination of sodium linear alkylbenzene sulfonate and sodium alkyl sulfate in carp.

Uptake, distribution, and elimination of labeled surfactants, 35S-labeled sodium laurylbenzenesulfonate (35S-C12-LAS) and 35S-labeled sodium lauryl sulfate (35S-C12-AS), were investigated in carp exposed to a low concentration level of the respective solutions found in river water in Japan by whole-body autoradiography and liquid scintillation counting. Whole-body autoradiograms showed that the fish absorbed 35S-C12-LAS and 35S-C12-AS from the solutions principally through gills and that the surfactants absorbed were rapidly distributed throughout the body tissues and organs. High concentrations were found in gills, blood, hepatopancreas, kidney, and gall bladder. The elimination in the fish that were kept in clean fresh water after exposure were relatively rapid in most tissues and organs except for gall bladder. These results suggest that the surfactants absorbed in fish may have an adverse effect on many tissues and organs of fish. What influenced the distribution profile and concentration factors of the surfactants in tissues and organs of carp were the exposure time and the molecular structure of the surfactants.     

Vertical distribution profiles of linear alkylbenzene sulfonates and their long-chain intermediate degradation products in coastal marine sediments

The variation with depth of the concentration of linear alkylbenzene sulfonates (LASs) and of the long-chain sulfophenyl carboxylic acids (SPCs) resulting from LAS biodegradation was determined in coastal sediments. We analyzed samples of sediment cores taken from three locations in a littoral zone subjected to the discharge of untreated urban effluents in the Bay of Cádiz in the southwestern part of the Iberian Peninsula. The vertical profile of LAS concentrations showed a sharp reduction with depth, whereas the concentration of long-chain SPCs (6-13 carbon atoms) was greatest at 10 to 14 cm depth. At this depth, the conditions in the interstitial water are strictly anoxic (Eh = -380 mV). The partition coefficients between the solid phase of the sediment versus the interstitial water are very different for LAS and for its degradation intermediates. For LAS, the organic carbon-based partition coefficient values were between 2.4 x 10(3) and 6.6 x 10(5)L/kg for the C10 and C13 homologues, respectively; these values are similar to those obtained from laboratory tests for the sorption of LAS onto marine sediments. For the long-chain SPCs, the partition coefficients are several orders of magnitude less as a consequence of their lower hydrophobicity.     

Comparative kinetics study of the evolution of freshwater aquatic toxicity and biodegradability of linear and branched alkylbenzene sulfonates

Evolution of both primary biodegradability and acute toxicity to daphnia and zebra fish of a linear alkylbenzene sodium sulfonate (LAS) and a branched alkylbenzene sodium sulfonate (BAS) have been measured simultaneously. In six of eight experiments, LAS was biodegraded to 90% in 7 days and BAS to 70% in 7 days. In the two other experiments, both LAS and BAS have shown the same biodegradation speed and reached the same biodegradation level in 7 days: 45% in one experiment and 55% in the other. The composition of bacteria population and the level of cellular ATP of the inoculum play a decisive role in the biodegradation. These results confirm that it is essential to know the composition of bacteria population present in the inoculum as well as their biochemical characteristics to accurately interpret results of biodegradation tests. In the case of a rapid primary biodegradation of LAS and BAS, the acute toxicity of LAS remains three times as high as that of BAS for at least 24 hr toward daphnia and 48 hr toward zebra fish. Their acute toxicity to daphnia and zebra fish become equivalent only after 72 hr. When primary biodegradation of both products is slower, the acute toxicity of LAS remains higher than that of BAS for more than 7 days.     

Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 3. Sublethal effects on soil invertebrates

Sewage sludge applied to agricultural soils often contains considerable amounts of linear alkylbenzene sulfonates (LAS). Toxic effects of LAS on soil organisms should, therefore. be evaluated to ensure safe use of sewage sludge as a fertilizer. In this study, dose-response relationships for the toxicity of Na-LAS to six species of soil invertebrates (survival, reproduction, and growth) were established using a sandy, agricultural soil as test substrate. In general, toxic effects on reproduction and growth appeared when the concentration in soil exceeded 40 to 60 mg/kg. Reproduction was approximately fourfold more sensitive in earthworms and enchytracids than in springtails and mites. It is argued that this difference in sensitivity is related to the dependency of soil pore water, which is high in the annelids but comparatively low in the arthropods.     

Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 1. Short-term effects on soil microbiology

Linear alkylbenzene sulfonates (LAS) may occur in sewage sludge that is applied to agricultural soil, in which LAS can be inhibitory to biological activity. As a part of a broader risk assessment of LAS in the terrestrial environment, we tested the short-term effects of aqueous LAS on microbial parameters in a sandy agricultural soil that was incubated for up to 11 d. The assays included 10 microbial soil parameters; ethylene degradation; potential ammonium oxidation; potential dehydrogenase activity; beta-glucosidase activity; iron reduction; the populations of cellulolytic bacteria, fungi and actinomycetes; the basal soil respiration; and the phospholipid fatty acid (PLFA) content. Except for beta-glucosidase activity, basal respiration, and total PLFA content, all soil parameters were sensitive to LAS, with EC10 values in the range of less than 8 to 22 mg/kg dry weight. This probably reflected a similar mode of LAS toxicity, ascribed to cell membrane interactions, and showed that sensitivity to LAS was common for various soil microorganisms. The extracellular beta-glucosidase activity was rather insensitive to LAS (ECI10, 47 mg/kg dry wt), whereas the basal soil respiration was not inhibited even at 793 mg/kg dry weight. This was interpreted as a combined response of inhibited and stimulated compartments of the microbial community. The PLFA content, surprisingly, showed no decrease even at 488 mg/kg. In conclusion, LAS inhibited specific microbial activities, although this could not be deduced from the basal respiration or the total PLFA content. The lowest EC10 values for microbial soil parameters were slightly higher than the predicted no-effect concentrations recently derived for plants and soil fauna (approximately 5 mg/kg dry wt).     

Dynamic in-stream fate modeling of xenobiotic organic compounds: a case study of linear alkylbenzene sulfonates in the Lambro River, Italy

Using a conceptual hydraulic model, a one-dimensional dynamic river water quality model has been developed to assess the short-term fate of linear alkylbenzene sulfonates (LAS) in the river compartments water and benthic sediment. The model assumes local equilibrium sorption and that both dissolved and sorbed chemical are available for biodegradation. To investigate the interaction of nutrient dynamics and organic contaminant fate, the model is coupled with a basic water quality model. On the basis of the Lambro River (Italy) as a case study, the result shows that the model predictions agree well with the measured data set. The model output sensitivity to model parameters has been tested, and the results depict that the model is highly sensitive to the biodegrading parameters. Also, a comparison of a steady state with a dynamic simulation and the effect of nutrient dynamics on the LAS fate in the Lambro River as a scenario analysis are presented. The results indicate the usefulness of the proposed model for the short-term simulation of organic contaminant fate in unsteady environmental conditions.     

Histological changes and copper accumulation in liver and gills of the Senegales sole, Solea senegalensis.

Senegales sole, Solea senegalensis, adult specimens were subjected to sublethal copper exposure (100 microg SO(4)Cu/L) for 7 days (contamination period) and afterward kept for 4 days without copper exposure (depuration period). In contaminated and detoxified specimens, the hepatocytes indicated a great number of lipid droplets compared to the control specimens. Some ultrastructural alterations such as partial disruption of the microvilli and endothelial lining of the sinusoids were observed in the hepatocytes. Lifting and swelling of the lamellar epithelium were common lesions in S. senegalensis gills due to the copper exposure. On the other hand, in contaminated and detoxified specimens, gills demonstrated separation of respiratory epithelium; chloride and pavement cells were also altered. Copper levels in gills, and especially in liver, increased during sublethal copper exposure (up to 7 days). During the detoxification period, Cu concentrations in gills and liver experienced variable changes. The results of the study indicate that histology is a successful tool capable of revealing sensitively and selectively even the sublethal effects of heavy metals on the environment and aquatic biota.     

Determination and distribution of alkyl ethoxysulfates and linear alkylbenzene sulfonates in coastal marine sediments from the Bay of Cadiz (southwest of Spain)

The distribution of the two main anionic surfactants manufactured and used in the world, alkyl ethoxysulfates (AES) and linear alkylbenzene sulfonates (LAS), has been studied in sediments from a salt marsh and an estuary of the Bay of Cadiz (southwest of Spain). The identification and quantification of AES and LAS was carried out after automated Soxhlet extraction with methanol, followed by solid-phase extraction and liquid chromatography coupled to electrospray mass spectrometry. The latter procedure permitted the unequivocal identification of every LAS homologue as well as the AES homologues of up to 16 carbon atoms in their alkyl chain and of up to 12 ethylene oxide groups. Recoveries were in the range of 51% to 84% and limits of detection from 1 to 5 microg/kg. We have focused our attention particularly on AES because, in spite of their great use, these compounds have received less attention than LAS and their occurrence has not been described in marine environmental samples. Alkyl ethoxysulfates concentration values range between 100 and 400 microg/kg in the topmost layer of sediments at the sampling areas. The relative distribution of AES homologues shows higher percentages for the longer alkyl chain homologues in sediments as well as for the shorter ethoxymers. A decrease in LAS concentrations has been found relative to past studies in one of the sampling areas as a consequence of the reduction of urban wastewater discharges.     

Uptake, elimination, and biotransformation of aqueous and dietary DDT in marine fish

To understand the biokinetics and potential risks of p,p'-dichlorodiphenyltrichloroethane (DDT) and its metabolites, p,p'-dichlorodiphenyldichloroethylene (DDE) and p,p'-dichlorodiphenyldichloroethane (DDD), in fish, we exposed the black sea breams (Acanthopagrus schlegeli) to aqueous and dietary DDTs and then evaluated the bioaccumulation, distribution, biotransformation, and elimination of DDTs under controlled laboratory conditions. The fish rapidly accumulated DDTs from both routes of exposure, particularly the gills and viscera. Elimination of DDTs following aqueous or dietary uptake was slow, and biotranslocation of DDTs was significant during the exposure period but negligible during the depuration period. The biotransformation process was more significant following dietary exposure. During depuration, DDE was the major biotransformed product in the sea breams' carcasses while DDD was the major product in the gills and viscera. However, DDD had a significantly higher elimination rate than DDE and, subsequently, the fish retained more DDE in the body. Intraspecies variability in the elimination and biotransformation processes in fish was observed. We demonstrated that the route of exposure significantly affected the fate and biokinetics of DDTs in fish. The application of a dynamic model provided a tool for quantifying the elimination and biotransformation of DDT in fish. The present study provided insights into the bioaccumulation and biotransformation pathways of DDT in fish that could have important ecotoxicological implications.     

Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 2. Effects on soil microbiology as influenced by sewage sludge and incubation time

The anionic surfactant linear alkylbenzene sulfonate (LAS) may inhibit soil microorganisms and may occur in agricultural soil through the application of sewage sludge. For five microbial parameters (microbial biomass C and the potentials of iron reduction, ammonium oxidation, dehydrogenase activity, and arylsulfatase activity), we compared the effects of aqueous LAS and LAS-spiked sewage sludge added to existing levels of 0, 3, 8, 22, 22, 62, 174, and 488 mg/kg soil (dry wt) in a Danish sandy agricultural soil that was incubated for 5 d to eight weeks. Arylsulfatase activity (measured after four weeks of incubation) was rather insensitive to LAS, with an EC 10 of 222 and more than 488 mg/kg in soil samples treated with aqueous LAS and LAS-spiked sewage sludge, respectively. For the other microbial parameters, the short-term effects (approximately one to two weeks) of aqueous LAS were characterized by an EC10 in the range of 3 to 39 mg/kg. Application of LAS via sewage sludge generally reduced the short-term effects for the microbial parameters, and the EC10 for LAS in sludge-amended soil after approximately one to two weeks of incubation ranged from less than 8 to 102 mg/kg. Recovery potential was seen for most microbial parameters as a result of prolonged incubation, both under conditions of LAS persistence (anaerobic conditions, the iron-reduction test) and LAS depletion (aerobic incubations, all other assays). In conclusion, the short-term inhibitory effects of LAS on soil microbiology were decreased in the presence of sewage sludge and by a prolonged (two to eight weeks) laboratory incubation period.     

Biodegradation kinetics of linear alkylbenzene sulfonate in sludge-amended agricultural soils

The kinetics of ultimate biodegradation (mineralization to CO2) of linear alkylbenzene sulfonate (LAS) were studied in sludge-amended agricultural soils for a series of pure chain length LAS homologs containing 10 to 14 carbon atoms in the alkyl chain. Degradation rates were measured by following the production of 14CO2 from uniformly 14C-ring-labeled material. In general, degradation of LAS was rapid in soil over a broad concentration range (0.1 to 10 times the expected environmental concentration) and demonstrated little variation among different homologs. Half-lives for mineralization of the benzene ring ranged from 18 to 26 days and were not significantly different for any homolog over the range of alkyl chain lengths tested. Half-lives measured for LAS degradation in these studies were comparable to values reported in the literature and also to values obtained for naturally occurring materials (stearic acid, cellulose) typically present in soil environments. On the basis of the results of the present studies and those of other investigators, it is concluded that soil environments exposed to LAS in sewage sludges contain microbial communities which can actively metabolize this material. Rates of biodegradation of the benzene ring, the final step in the LAS biodegradation pathway prior to complete mineralization, are also sufficient to prevent LAS from accumulating in soil environments.     

Atrazine uptake, elimination, and bioconcentration by periphyton communities and Daphnia magna: effects of dissolved organic carbon

The bioconcentration and toxicokinetics of atrazine in three different periphyton communities and in laboratory reared Daphnia magna were studied in natural and artificial waters with different dissolved organic carbon (DOC) concentrations and qualities. The exposure concentrations were similar to those short-lived peak concentrations found in contaminated waters. Atrazine uptake and elimination were very fast, and the bioconcentration was low both in periphyton and D. magna. The bioconcentration factors in D. magna were approx. 16% of those in periphyton. The uptake and elimination rates were also higher in periphyton than in Daphnia. The periphyton properties affected the bioconcentration of atrazine more than the DOC concentration and water quality. A steady state was reached within a few hours of uptake by the periphyton and within 24 h by D. magna. A residue of atrazine was left in the periphyton and D. magna after an 8-h biphasic depuration with clearly distinct fast and slow phases. In D. magna, atrazine may be eliminated during the normal life span of the organism. These results suggest that DOC has a minor influence on the toxicokinetics of atrazine.     

Dissolved organic matter modulating the uptake, biotransformation, and elimination rates of pyrene in Daphnia magna

In the current study, the toxicokinetics of pyrene in Daphnia magna was determined in detail, and the role of dissolved organic matter (DOM) in these processes was investigated. Uptake, biotransformation, parent compound and metabolite elimination rates (k(u), k(m), k(ep), k(em)), depuration rates, bioconcentration factors (BCFs), and biological half-lives were studied in one natural humic water, in two waters made from DOM isolates, and in DOM-free artificial fresh water. The daphnids were exposed to pyrene for 24 h and depurated for another 24 h. To analyze the data, the two-compartment model was used to obtain toxicokinetic parameters, and the depuration data were analyzed by linear regression. Extensive biotransformation of pyrene resulted in high metabolite concentration in the organisms (83% of total radioactivity) after 24 h exposure. Depuration of the parent compound seemed to be biphasic: at first, depuration was rapid, but after 4 to 10 h in uncontaminated water, the disappearance rate came close to zero. After a 24-h depuration time, 7.8% of the pyrene accumulated remained in the organism. Overall, the elimination of parent compound was faster than the elimination of metabolites, which might imply inefficient excretion of water-soluble biotransformation products from daphnids. The effect of DOM on accumulation of pyrene was seen in a decreased uptake rate, indicating lower bioavailability of the compound in waters that contain high-aromaticity DOM. In the same waters, DOM significantly accelerated pyrene biotransformation and overall elimination. Altogether, DOM appears to be able to affect toxicokinetics through mechanisms that are connected to interaction with the contaminant but also directly through interaction with biotransformation systems.     

Influence of soil moisture on linear alkylbenzene sulfonate-induced toxicity in ammonia-oxidizing bacteria

Moisture affects bioavailability and fate of pollutants in soil, but very little is known about moisture-induced effects on pollutant toxicity. We here report on a modifying effect of moisture on degradation of linear alkylbenzene sulfonates (LASs) and on their toxicity towards ammonia-oxidizing bacteria (AOB) in agricultural soil. In soil spiked with two LAS levels (250 or 1,000 mg/kg) and incubated at four different moisture levels (9-100% of water-holding capacity), degradation was strongly affected by both soil moisture and initial LAS concentration, resulting in degradation half-lives ranging from 13 to more than 160 d. Toxicity towards AOB assessed by a novel Nitrosomonas europaea luxAB-reporter assay was correlated to total LAS concentration, indicating that LAS remained bioavailable over time without accumulation of toxic intermediates. Toxicity towards indigenous AOB increased with increasing soil moisture. The results indicate that dry soil conditions inhibit LAS degradation and provide protection against toxicity within the indigenous AOB, thus allowing for a rapid recovery of this population when LAS degradation is resumed and completed after rewetting. We propose that the protection of microbial populations against toxicity in dry soil may be a general phenomenon caused primarily by limited diffusion and thus a low bioavailability of the toxicant.     

Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 4. The influence of salt speciation, soil type, and sewage sludge on toxicity using the collembolan Folsomia fimetaria and the earthworm Aporrectodea caliginosa as test organisms

Sewage sludge applied to agricultural soils often contains linear alkylbenzene sulfonates (LAS) in the range of 1 to 10 g/kg dry weight, and their toxicity to relevant soil organisms should, therefore, be assessed to ensure safe use of sewage sludge as a fertilizer. Studies of LAS toxicity to soil organisms are few, and to our knowledge, factors that may influence the toxicity in the field have not been studied in detail. In this paper, we report on the influence of speciation of LAS in the test solution added to soil (soluble Na-LAS vs poorly soluble Ca-LAS or Mg-LAS), the influence of soil type, and the modifying effects of sludge amendment on the toxicity of LAS. These issues were investigated using reproduction of Collembola and growth of juvenile earthworms as test parameters. Speciation of the LAS added to test soil did not have any influence on toxicity for any of the test species. Likewise, in three different agricultural soils (sand, loam, and clay), we found almost equal toxicities. The LAS added to test soil in a sludge-water suspension was equally toxic as when it was added in an aqueous solution. However, anaerobic incubation for 7 and 14 d of the LAS-sludge suspension (with no decay of LAS) caused the toxicity to increase almost threefold in both collembolan and earthworm. The relationships between soil constituents, bioavailability, and toxicity are also discussed.