Influence of water hardness on the bioavailability and toxicity of linear alkylbenzene sulphonate (LAS)

In the present work, the influence of Ca²⁺ concentration on the toxicity of single cut linear alkylbenzene sulfonate (LAS) homologues was studied. Precipitation boundary diagrams for each homologue were obtained, indicating turbid and clear zones depending on the LAS and Ca²⁺ concentrations. The separation between transparent and turbid zones is given by the so-called precipitation line. LAS toxicity to Daphnia magna was determined at concentrations close to this precipitation line. It was observed that when Ca(LAS)₂ precipitation progresses, LAS bioavailability decreases for test animals, and the toxicity diminishes even at high nominal LAS concentrations. According to the “free ion activity model” (FIAM), the toxicity of a given chemical compound is mainly due to the ionic species (Ca²⁺–LAS⁻) and not due to the precipitated molecule, Ca(LAS)₂. The significance of the present study is in connection with the assessment of LAS sorption/precipitation studies in soils and sediments, where in situ toxicity is strongly influenced by Ca²⁺/Mg²⁺ ions, according to the results presented in this work.     

Effect of linear alkylbenzene sulphonate (LAS) on the mineralization, metabolism and uptake of 14C-phenanthrene in a model ecosystem (water-lava-plant-air)

The aim of this work was to evaluate the effect of linear alkylbenzene sulfonate (LAS, 200 mg l(-1)) on the fate of phenanthrene in a model ecosystem "water-lava-hydrophytes-air". The experiments were conducted using two closed cultivation chamber systems. Rushes (Juncus effesus) were selected as a representative hydrophyte. Five hundred micrograms per liter of phenanthrene in a culture solution containing a 14C-activity of 75 microCi per chamber was applied (i) to investigate the degradation of the labeled test substance and the transfer processes within the system; (ii) to determine the mass-balance possible and (iii) to detect the occurrence of volatile test substances, their volatile metabolites and the degradation end-product CO2 in the gas phase. Most of the applied 14C-activity was found in the plant (41-45%), in which approximately 95% was associated with plant roots and approximately 5% with shoots. The 14C-activity recovered in the form of VOCs and CO2 was measured in lava (18-29%, 8-11%), and in the culture solution (10-14% and 1%), respectively. Majority of the applied 14C-activity existed in two forms, i.e. (1) polar metabolites (26%), of which 91% were found in plant roots, and (2) un-extractable residues (23%), most of which were in plant roots (40%) and bounded to lava (58%). The presence of LAS significantly increased the volatilization of phenanthrene and its metabolites, inhibited its mineralization and decreased the level of 14C-activity in lava. Moreover, LAS reduced the phenanthrene level in plant roots.     

Chronic toxicity of sediment-associated linear alkylbenzene sulphonates (LAS) to freshwater benthic organisms

The toxicity of linear alkylbenzene sulphonates (LAS), to freshwater benthic organisms was assessed during exposure to spiked sediment. Lethal and sub-lethal end-points were monitored for two organisms (oligochaete Lumbriculus variegatus and nematode Caenorhabditis elegans). Results demonstrated relatively low toxicity (LOECs>100 mg/kg dry weight). No observed effect concentrations (NOECs) of 81 mg/kg dw (Lumbriculus) and 100 mg/kg dw (Caenorhabditis) were determined. For the oligochaete, no specific endpoint was particularly sensitive to LAS. For the nematode, egg production was the most sensitive endpoint. Significant degradation was measured over the 28-day duration of the Lumbriculus study, equating to a half-life of 20 days in sediment.     

Risk assessment of linear alkylbenzene sulphonates, LAS, in agricultural soil revisited: robust chronic toxicity tests for Folsomia candida (Collembola), Aporrectodea caliginosa (Oligochaeta) and Enchytraeus crypticus (Enchytraeidae)

To obtain robust data on the toxicity of LAS, tests with the collembolan Folsomia candida L., the oligochaetes Aporrectodea caliginosa Savigny (earthworm) and Enchytraeus crypticus Westheide and Graefe (enchytraeid) were performed in a sandy loam soil. Additionally limited tests with LAS spiked to sewage sludge, and subsequently mixed into soil, were performed. For the endpoint of interest, reproduction in soil, we found an EC10 of 205 mg LAS kg(-1) soil [8.6-401] [95% confidence limits] for F. candida and an EC10 of 46 mg LAS kg(-1) soil [13-80] for A. caliginosa after 28 days. E. crypticus was not affected by concentrations up to 120 mg LAS kg(-1) soil. When adding (low contaminated) non-spiked sludge to soil, high stimulation of reproduction was observed for E. crypticus and A. caliginosa but not for F. candida. We argue that this difference in stimulative response between the tested species is related to the difference in feeding behaviour. Sludge spiked with LAS did not significantly affect the reproduction of F. candida (fertility: number of juvenile offspring) and A. caliginosa (fecundity: number of cocoons) (dose equivalent to 181 g and 91 g LAS kg(-1) sludge, respectively). Significantly reduced reproduction was observed for E. crypticus (at 120 mg LAS kg(-1) soil+sludge corresponding to 72 g LAS kg(-1) sludge) compared to non-spiked sludge. The reproduction by E. crypticus was, however, comparable to the reproduction observed in the control soil without sludge. Compared to LAS directly spiked to soil, the reproductive output of organisms exposed to spiked sludge was either not significantly different (F. candida, E. crypticus) or significantly improved (A. caliginosa). More studies are needed in order to make firm conclusions on the potential effect of artificially contaminated sludge in soil systems.     

Effect of introduced phthalate-degrading bacteria on the diversity of indigenous bacterial communities during di-(2-ethylhexyl) phthalate (DEHP) degradation in a soil microcosm

Four previously isolated di-butyl-phthalate (DBP) degraders were tested for their abilities to degrade di-(2-ethylhexyl) phthalate (DEHP). In aqueous medium supplemented with 100mg/l of DEHP, both isolate G1 and Rhodococcus rhodochrous G2 showed excellent degradative activity; in three days they were able to degrade more than 97% of the added DEHP. Rhodococcus rhodochrous G7 degraded 32.5% of the added DEHP and Corynebacterium nitrilophilus G11 showed the least amount of DEHP degradation. The addition of surfactant Brij 30 at 0.1x critical micelle concentration (2mg/l) significantly improved DEHP degradation by Rhodococcus rhodochrous G2 (more than 90% of the added DEHP was degraded within 24 hours), but slightly inhibited the degradation of DEHP by the isolate G1 and Rhodococcus rhodochrous G7. Based on the 16S rDNA sequence data, isolate G1 was identified as Gordonia polyisoprenivorans. Soil inhibited DEHP degradation by G. polyisoprenivorans G1; fourteen days after a second addition of DEHP, 11.5% of the total added DEHP (i.e., 243.4 microg/g soil) remained detectable. Changes in the bacterial community were monitored using denaturing gradient gel electrophoresis (DGGE) and respective dendrogram analysis. It is clear that DEHP and DEHP plus G. polyisoprenivorans G1 substantially affected the bacterial community structure in the soils. However, as the population of indigenous DEHP degraders increased in the DEPH-treated soil, its bacterial communities resembled those in the DEHP plus G. polyisoprenivorans G1-inoculated soil by Day 17.     

In vitro biotransformation of surfactants in fish. Part I: linear alkylbenzene sulfonate (C12-LAS) and alcohol ethoxylate (C13EO8)

Developing regulatory activities (e.g., REACh, [DGEE. 2003. Directorates General Enterprise and Environment. The new EU chemicals legislation REACH. DG Enterprise, Brussels, Belgium. (http://www.europa.eu.int/comm/enterprise/reach/index_en.htm)]) will require bioaccumulation to be assessed for thousands of chemicals. Further, there is increasing pressure to reduce, refine or replace animal tests. Given this scenario, there is an urgent need to evaluate the feasibility of in vitro systems to supply data useful for bioaccumulation estimation. Subcellular and cellular hepatic systems were tested to determine the biotransformation of two surfactants: C12-2-LAS (2-phenyl dodecane p-sulfonate) and an alcohol ethoxylate C13EO8 (Octaethylene glycol monotridecyl ether). The subcellular systems tested were liver homogenates and microsomes from the common carp (Cyprinus carpio) and rainbow trout (Oncorhynchus mykiss). Cellular systems consisted of primary hepatocytes from the common carp (Cyprinus carpio) and PLHC-1 cells, hepatocarcinoma cells from the desert topminnow (Poeciliopsis lucida). All in vitro systems were exposed to radiolabeled test compounds and assayed for biotransformation using liquid scintillation and thin layer chromatographic methods. First-order kinetics were used to estimate rates of biotransformation. Bioconcentration of test materials in fish were predicted using an in vitro to in vivo metabolic rate extrapolation model linked to a mass-balance model commonly used to predict bioaccumulation in fish. Subcellular biotransformation rates for each of the surfactants were greatest with microsomes. Cellular loss rates exceeded subcellular rates, leading to lower predicted BCF values. Predicted BCFs corresponded closely to measured values in several fish species, verifying the utility of in vitro systems in refining Kow-only-based BCFs via the inclusion of biotransformation rates.     

Nonylphenols, nonylphenol-ethoxylates, linear alkylbenzenesulfonates (LAS) and bis (4-chlorophenyl)-sulfone in the German Bight of the North Sea.

Nonylphenols and nonylphenol-ethoxylates were detected in the water and sediment samples from the German Bight of the North Sea. Additionally bis (4-chlorophenyl)-sulfone and linear alkylbenzenesulfonates (LAS) were detected in marine waters. Proof of identification is given by comparison of spectral and chromatographic data from the compounds in sample extracts to those obtained from pure standards. In extracts obtained from water samples taken in 1990 and 1995 the concentrations of nonylphenols and bis-(4-chlorophenyl)-sulfone were compared for each year. The concentrations of nonylphenols in seawater varied from 0.7 to 4.4 ng/l while in the Elbe estuary about 33 ng/l were found. In water samples taken in 1998 nonylphenol-polyethoxylates could not be determined, whereas LAS concentrations of 30 ng/l were confirmed by HPLC-MS/MS. The concentrations of bis (4-chlorophenyl)-sulfone ranged from 0.18 to 2.2 ng/l. In sediment samples LAS concentrations of 39-109 ng/g dry weight were determined.     

Studies on the restoration succession of PFU microbial communities in a pilot-scale microcosm

In order to imitate the restoration succession process of natural water ecosystem, a laboratory microcosm system of constant-flow-restoration was designed and established. A eutrophycation lake, Lake Donghu, was selected as the subject investigated. Six sampling stations were set on the lake, among which the water of station IV was natural clean water, and others were polluted with different degrees. Polyurethane foam unit microbial communities, which had colonized in the stations for a month, were collected from these stations and placed in their respective microcosms, using clean water of station IV to gradually replace the water of these microcosms. In this process, the healthy community in clean water continuously replaced the damaged communities in polluted water, the restoration succession of the damaged communities was characterized by weekly determination of several functional and structural community parameters, including species number (S), diversity index (DI), community pollution value (CPV), heterotrophy index (HI), and similarity coefficient. Cluster analysis based on similarity coefficient was used to compare the succession discrepancies of these microbial communities from different stations. The ecological succession of microbial communities during restoration was investigated by the variable patterns of these parameters, and based on which, the restoration standards of these polluted stations were suggested in an ecological sense. That was, while being restored, the water of station 0 (supereutrophycation) should be substituted with natural clean water by 95%; station I (eutrophycation), more than 90%; station II (eutrophycation), more than 85%; station III (eutrophycation), about 85%; station V (mesoeutrophycation), less than 50%. The effects of the structural and functional parameters in monitoring and assessing ecological restoration are analyzed and compared.     

Fate of Bacillus thuringiensis (Bt) Cry3Bb1 protein in a soil microcosm

Transgenic crops expressing insecticidal, crystalline (Cry) Bacillus thuringiensis (Bt) proteins were commercialized in the US in 1996. There is little information in the peer-reviewed literature on the environmental fate of the coleopteran-active Bt Cry3Bb1 protein expressed in event MON863 corn. The exposure characterization of Bt proteins is unique in that the fate of the protein in soil and in crop residue must be considered. To date, the significance of macrodecomposing organisms, such as earthworms, isopods, and springtails, to the dissipation of Bt proteins present in crop residue has not been assessed. In addition, the input of Bt proteins into soil through leaching from post-harvest crop residue has not been examined. Two laboratory microcosm studies were conducted to determine the fate of Bt Cry3Bb1 in decomposing MON863 corn residue and to determine whether Bt proteins can enter soil by leaching from crop residue. In addition, the importance of macrodecomposing organisms to the degradation of Bt proteins in corn residue was assessed. Laboratory microcosms containing MON863 corn leaf, root, and stalk with and without macrodecomposers were used to examine the fate of Bt Cry3Bb1 in post-harvest MON863 corn residue. A half-life of less than five days was found for Bt Cry3Bb1 protein in decomposing MON863 corn residue. There was a trend of increasing half-life of Cry3Bb1 in microcosms containing macrodecomposers, however, this trend was only significant (p<0.05) for Bt Cry3Bb1 in MON863 leaf tissue and this increase is not likely relevant for non-target exposure. The recovery of Bt Cry3Bb1 protein from soil extracts was either below the limit of quantification (9 ng g(-1) soil) or below the limit of detection (0.7 ng mL(-1)) at all time points during the study. Based on the results from this study, Bt protein leaching from post-harvest crop residue is not a significant contributor to Bt protein input into soil.     

Polychlorinated biphenyl (PCB) anaerobic degradation in marine sediments: microcosm study and role of autochthonous microbial communities

Polychlorobiphenyl (PCB) biodegradation was followed for 1 year in microcosms containing marine sediments collected from Mar Piccolo (Taranto, Italy) chronically contaminated by this class of hazardous compounds. The microcosms were performed under strictly anaerobic conditions with or without the addition of Dehalococcoides mccartyi, the main microorganism known to degrade PCBs through the anaerobic reductive dechlorination process. Thirty PCB congeners were monitored during the experiments revealing that the biodegradation occurred in all microcosms with a decrease in hepta-, hexa-, and penta-chlorobiphenyls (CBs) and a parallel increase in low chlorinated PCBs (tri-CBs and tetra-CBs). The concentrations of the most representative congeners detected in the original sediment, such as 245-245-CB and 2345-245-CB, and of the mixture 2356-34-CB+234-245-CB, decreased by 32.5, 23.8, and 46.7 %, respectively, after only 70 days of anaerobic incubation without any bioaugmentation treatment. Additionally, the structure and population dynamics of the microbial key players involved in the biodegradative process and of the entire mixed microbial community were accurately defined by Catalyzed Reporter Deposition Fluorescence In Situ Hybridization (CARD-FISH) in both the original sediment and during the operation of the microcosm. The reductive dehalogenase genes of D. mccartyi, specifically involved in PCB dechlorination, were also quantified using real-time PCR (qPCR). Our results demonstrated that the autochthonous microbial community living in the marine sediment, including D. mccartyi (6.32E+06 16S rRNA gene copy numbers g^?1 sediment), was able to efficiently sustain the biodegradation of PCBs when controlled anaerobic conditions were imposed.     

Adsorption/desorption of linear alkylbenzenesulfonate (LAS) and azoproteins by/from activated sludge flocs

Our study investigated the adsorption/desorption by/from activated sludge flocs, dispersed in river water or in diluted wastewater, of organic compounds (C(11)-LAS, azoalbumin and azocasein) at concentrations relevant to environmental conditions. Activated sludge flocs, used as a model of biological aggregates, are characterized by a very heterogeneous matrix able to sorb the three organic compounds tested at 4 degrees C. The adsorbed amount of C(11)-LAS by activated sludge flocs was higher than that of azocasein or azoalbumin, as shown by the Freundlich parameters (K(ads)=8.6+/-1.7, 1.6+/-0.3 and 0.3+/-0.1 micromol(1-1/n)g(-1)l(1/n) for C(11)-LAS, azocasein and azoalbumin, respectively; n=3 sludges). C(11)-LAS sorption from activated sludge appeared to be partially reversible in river water, while a marked hysteresis phenomenon was observed for azocasein and azoalbumin, implying a low degree of reversibility in their exchange between activated sludge and river water. It has also been displayed that the conductivity variation of bulk water (comprised between 214 and 838 microS cm(-1)) exerted no dramatic effect on the C(11)-LAS desorption from activated sludge flocs, while a little effect of it on azocasein desorption was observed. Thus, biological aggregates as activated sludge flocs can serve as an intermediate carrier for C(11)-LAS, while it represents a sink for proteins.     

Effects of a non-ionic surfactant (Tween-80) on the mineralization, metabolism and uptake of phenanthrene in wheat-solution-lava microcosm

Effects of a non-ionic surfactant (Tween-80) on the mineralization, metabolism and uptake of phenanthrene in wheat-solution-lava microcosm were studied using 14C-labeled phenanthrene. The mineralization and metabolism of phenanthrene were fast in such a system. At least 90% of the applied phenanthrene were transformed within 24 days. Only 0.3% of the applied 14C-activity were identified to be the parent phenanthrene. Most of the applied 14C-activity (70%) was recovered from wheat, in which ca. 70% were associated with wheat shoots (stems and leaves) and ca. 30% wheat roots. 33% and 20% of the applied 14C-activity had been constructed into wheat tissues of shoots and roots, respectively. The 14C-activity recovered in forms of CO2 and volatile organic chemicals (VOCs) was 12-16% and 4-5%, respectively. The major metabolites of phenanthrene were polar compounds (18% of the applied 14C) and only 2.1% were identified as non-polar metabolites. No phenanthrene was found in wheat shoots indicating that it could not be transported from roots to upper parts of the plant but in form of metabolites (mostly polar metabolites). Foliar uptake of 14C-activity via air in form of 14CO2 occurred. The presence of Tween-80 significantly enhanced the degradation of phenanthrene, which could be attributed to its increase of microbial activities in the system. Tween-80 also significantly (P < 0.05) reduced the phenanthrene level in wheat roots, which probably resulted from desorption of phenanthrene from root surface caused by the surfactant.     

Characterisation of perfluorooctane sulfonate (PFOS) in a terrestrial ecosystem near a fluorochemical plant in Flanders,Belgium

Bioaccumulation of perfluorooctane sulfonate (PFOS) in a restricted terrestrial food chain was investigated with the omnivorous wood mouse (Apodemus sylvaticus) on top of the studied food chain. The levels detected are very high compared with literature as a result of the presence of fluorochemical plant in the immediate vicinity of the study area. Soil, surface water, fruits of European elder and common blackberry, invertebrates, bank vole and wood mouse were collected at two sites, e.g. Blokkersdijk, adjacent to the fluorochemical plant, and Galgenweel, a reference site 2 km further away. In wood mouse, the highest PFOS concentrations were found in the liver followed by the pancreas, lungs and kidneys, with the spleen having the lowest levels. In the liver, the concentrations ranged from 787 to 22,355 ng/g ww at Blokkersdijk and these were significantly correlated with those detected in the kidneys (13.7–4,226 ng/g ww). If current results are compared to the findings of a previous study conducted in 2002 at the same sites, a significant decrease of PFOS in livers of wood mouse is observed. To the best of our knowledge, so far no studies reported levels of PFOS in terrestrial invertebrates under field conditions. At Blokkersdijk, PFOS was detected in all invertebrate species ranging from 28 to 9,000 ng/g. Soil and water were also contaminated with levels of respectively 68 ng/g and 22 ng/L. Biota-to-soil accumulation factors ranged from 0.11 to 68 for earthworms. Biomagnification factors (BMFs) of liver wood mouse/berries were as high as 302. BMFs for invertebrates were remarkably lower (up to 2).     

Influence of lead acetate on soil microbial biomass and community structure in two different soils with the growth of Chinese cabbage (Brassica chinensis)

A greenhouse pot experiment was conducted to evaluate the impact of different concentrations of lead acetate on soil microbial biomass and community structure during growth of Chinese cabbage (Brassica chinensis) in two different soils. The field soils were used for a small pot, short-term 60-day growth chamber study. The soils were amended with different Pb concentrations, ranging from 0 to 900mgkg(-1) soil. The experimental design was a 2 soilx2 vegetation/non-vegetationx6 treatments (Pb)x3 replicate factorial experiment. At 60 days the study was terminated and soils were analyzed for microbial parameters, namely, microbial biomass, basal respiration and PLFAs. The results indicated that the application of Pb at lower concentrations (100 and 300mgkg(-1)) as lead acetate resulted in a slight increase in soil microbial biomass, whereas Pb concentrations >500mgkg(-1) caused an immediate gradual significant decline in biomass. However, the degree of impact on soil microbial biomass and basal respiration by Pb was related to management (plant vegetation) or the contents of clay and organic matter in soils. The profiles of 21 phospholipid fatty acids (PLFAs) were used to assess whether observed changes in functional microbial parameters were accompanied by changes in the composition of the microbial communities after Pb application at 0, 300 and 900mg Pbkg(-1) soil. The results of principal component analyses (PCA) indicated that there were significant increases in fungi biomarkers of 18:3omega6c, 18:1omega9c and a decrease in cy17:0, which is an indicator of gram-negative bacteria for the high levels of Pb treatments In a word, soil microbial biomass and community structure, therefore, may be sensitive indicators reflecting environmental stress in soil-Pb-plant system. However, further studies will be needed to better understand how these changes in microbial community structure might actually impact soil microbial community function.     

Biodegradation of PAHs in soil: Influence of chemical structure, concentration and multiple amendment

The influence of PAH chemical structure and concentration, added in either single (75 or 300 mg kg⁻¹) or multiple (2 × 75, 2 × 150 or 4 × 75 mg kg⁻¹) applications as single- or multiple-contaminant systems, on the development of PAH biodegradation in a pristine soil was investigated. Development in microbial catabolic ability was assessed at 0, 28, 56 and 84 d by monitoring ¹⁴C-naphthalene, ¹⁴C-phenanthrene and ¹⁴C-pyrene mineralisation over 14 d in respirometric assays. The presence of other contaminants influenced the ability of the indigenous microflora to mineralise structurally different contaminants over time. ¹⁴C-Naphthalene mineralisation was inhibited by the presence of other contaminants; whereas the presence of naphthalene significantly enhanced rates of mineralisation in multiple-contaminant systems containing ¹⁴C-phenanthrene and ¹⁴C-pyrene. Generally, increasing the number of contaminant applications has implications for catabolic activity of soil microbes. It is suggested the toxic nature of PAHs retarded mineralisation at increased contaminant concentrations.     

A comparative study on sorption of perfluorooctane sulfonate (PFOS) by chars, ash and carbon nanotubes

Perfluorooctane sulfonate (PFOS), as one of emerging contaminants, has been attracting increasing concerns in recent years. Sorption of PFOS by maize straw- and willow-derived chars (M400 and W400), maize straw-origin ash (MA) as well as three carbon nanotubes (CNTs) was studied in this work. The sorption kinetics of PFOS by the six adsorbents was well fitted by the pseudo-second-order model. CNTs reached equilibrium in 2 h, much faster than those by chars (384 h) and ash (48 h). According to the sorption isotherms, both single-walled carbon nanotubes (SWCNT) and MA had high sorption capacities (over 700 mg g⁻¹), while the two chars had low sorption capacities (below 170 mg g⁻¹) caused by their small BET surface area. In the case of MA, due to its positively charged surface, both hydrophobic interaction and electrostatic attraction involved in the sorption, and the formation of hemi-micelles further favored the sorption. This study suggested that SWCNT and MA were effective adsorbents for PFOS removal from water. Compared to SWCNT, MA is low cost and easy to obtain, so it could be a preferred adsorbent for PFOS removal.     

Assessment of natural sepiolite on cadmium stabilization, microbial communities, and enzyme activities in acidic soil

A pot trial was conducted to assess the efficiency of sepiolite-induced cadmium (Cd) immobilization in ultisoils. Under Cd concentrations of 1.25, 2.5, and 5 mg?kg^?1, the available Cd in the soil after the application of 1–10 % sepiolite decreased by a maximum of 44.4, 23.0, and 17.0 %, respectively, compared with no sepiolite treatments. The increase in the values of soil enzyme activities and microbial number proved that a certain metabolic recovery occurred after sepiolite treatment. The dry biomass of spinach (Spinacia oleracea) increased with increasing sepiolite concentration in the soil. However, the concentration (dry weight) of Cd in the spinach shoots decreased with the increase in sepiolite dose, with maximum reduction of 92.2, 90.0, and 84.9 %, respectively, compared with that of unamended soils. Under a Cd level of 1.25 mg?kg^?1, the Cd concentration in the edible parts of spinach at 1 % sepiolite amendment was lower than 0.2 mg?kg^?1 fresh weight, the maximum permissible concentration (MPC) of Cd in vegetable. Even at higher Cd concentrations (2.5 and 5 mg?kg^?1), safe spinach was produced when the sepiolite treatment was up to 5 %. The results showed that sepiolite-assisted remediation could potentially succeed on a field scale by decreasing Cd entry into the food chain.