Soil S availability in upland pastures of NE Scotland: relationship of extractable soil S and soil respiration to soil and site characteristics

Abstract. The mean extractable sulphur (S) concentration in 315 upland topsoil samples collected in 1988/89 from beneath pasture in NE Scotland was 13 μg S g⁻¹ (range 2–77 μg S g⁻¹). More than two thirds of the samples had S concentrations less than that acceptable for productive soils. Continued decreases in atmospheric S inputs may have increased this proportion subsequently. The analysis of herbage S also indicated that two-thirds of the samples were below 0.2% S. A 'respirometric index', namely CO₂ produced during cellulose decomposition without added S as a percentage of that produced with added S, was significantly less than 100% in a quarter of the soils. Results of three different extraction procedures suggested that sulphate in the soils was present mainly as free plus adsorbed rather than precipitated forms. Soil extraction identified a significant non-sulphate S fraction, presumably organic S. The variability in extractable S stemmed from a combination of geographical, depositional and local site and soil factors. Extractable S was significantly correlated with soil organic matter content and inversely with soil pH and together these factors explained 37% of the variability. While significant differences in mean concentrations between geographical area, soil association and drainage status were evident, no trends could be observed between the major soil subgroups or with altitude.     

Quantitative determination of dietary lectin activities by enzyme-linked immunosorbent assay using specific glycoproteins immobilized on microtiter plates

An immunoenzymatic method for the quantitative determination of dietary lectin activities employing immobilized glycoproteins was studied. Lectins from wheat germ (WGA), peanut (PNA), and jack bean (ConA) were added to microtiter plates coated with ovalbumin or asialofetuin and quantified by enzyme-linked immunosorbent assay (ELISA) with lectin-specific antibodies. ELISA responses for lectin activity were dose-dependent in the concentration range 30-1000 ng/mL for WGA and 80-1000 ng/mL for both PNA and ConA. Inhibition assays carried out with different saccharides confirmed that the binding of lectins to immobilized glycoproteins was specific. The proposed method is specific and sensitive, allowing the quantitative determination of lectin activities on raw samples by simple dilution of the extracts. Examples of application to wheat germ and roasted peanut extracts are reported.     

Runoff and leaching of alachlor under conventional and soil-specific management

Abstract. The influence of conventional and soil-specific management on leaching and runoff losses of soil-applied alachlor (2-chloro-2',6'-diethyl- N -(methoxymethyl) acetanilide) was studied across a soil catena (landscape) with varied slope and drainage characteristics. The catena consisted of: a well-drained Ves (fine-loamy, mixed, mesic Udic Haplustoll) soil on the backslope (1–4%), a Ves soil on the sideslope (6–12%), and a poorly drained Webster (fine-loamy, mixed, mesic Typic Haplaquoll) soil on the toeslope (0–3%). In general, the concentration of alachlor in runoff water was greater in the Ves soil than in the Webster. In 1992 alachlor concentrations in runoff (water, sediment + water) were less for soil-specific rates (2.20 or 2.80 kg/ha) than for a uniform rate (3.36 kg/ha) in both Ves soils. There was no significant difference in alachlor concentration related to application rates (soil-specific rate 3.66 kg/ha) in the runoff from the Webster soil. Averaged across soils and events, the concentrations of alachlor in runoff (water, sediments + water) were less for soil-specific rates than for the uniform rate. Alachlor was not detected in soil samples obtained from depths greater than 15 cm in any soil or treatment after the first sampling. At the first sampling in 1992 (7 days after application) alachlor was detected down to 45 and 90 cm in the Ves and Webster soils, respectively. Detectable amounts (≥0.1 μg/1) of alachlor were observed in soil water samples extracted from all three soils during some sampling dates. No particular trends were observed with soils or application rates.     

Mapping the distribution of DDT residues as DDE in the soils of the irrigated regions of northern New South Wales,Australia using ELISA and GIS

An enzyme-linked immunosorbent assay (ELISA) specific for DDE [1,1-dichloro-2,2-bis(4-chlorophenyl)ethylene] has been used to map DDT [1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane)] residues in the top 10 cm of soil in three river valleys of northern New South Wales, Australia. Despite being almost 20 years since DDT was last applied for cotton growing in these areas, the relationship between sites of greatest application and current residue levels was strong. DDE concentrations in the range 0-2 ppm were found, although most the 389 soil samples examined contained less than 0.2 ppm of DDE. Although some relationship between mode of land use and current residue levels was apparent, this varied from valley to valley and may have reflected different farming practices and times of application. The study demonstrates that the combination of ELISA and geographical information system (GIS) analysis provides an effective means of displaying levels of soil contamination by a pesticide and the possible need for remediation.     

Application of an enzyme-linked immunosorbent assay for the analysis of imidacloprid in wiliwili tree, Erythrina sandwicensis O. Deg, for control of the wasp Quadrastichus erythrinae

A monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) for the neonicotinoid insecticide imidacloprid was evaluated for its reproducibility, accuracy, and comparability to results from a conventional high-performance liquid chromatography (HPLC) for the analysis of imidacloprid in the endemic wiliwili tree (Erythrina sandwicensis O. Deg) found in dryland forests and landscapes in Hawaii. Imidacloprid was applied to these wiliwili trees in an attempt to control the newly introduced erythrina gall wasp, Quadrastichus erythrinae Kim. Leaf samples were freeze-dried and extracted with acidic aqueous methanol followed by methylene chloride partitioning. After solvent removal, the extract residue was reconstituted in 1 mL of water/methanol (1:1, v/v) for ELISA; no significant matrix interference was observed at 10-fold or more dilution. The average recoveries of imidacloprid from fortified samples ranged from 78% to 100% by ELISA. The correlation between the ELISA and HPLC results was excellent (r2 = 0.98). Imidacloprid was detected with the ELISA in all treated samples and its level varied in the samples among different treatments and in those from different parts of the trees. The infestation severity rating of leaf samples was inversely related to the concentration of imidacloprid. It is clear that imidacloprid effectively controls the wasps. The ELISA is a suitable method for quantitative and reliable determination of imidacloprid in wiliwili trees and the application provides information to understand how to control the wasps.     

Poly- and monoclonal antibody-based ELISAs for fipronil

An enzyme-linked immunosorbent assay (ELISA) for fipronil was developed by using polyclonal antibodies (pABs) or monoclonal antibodies (mABs), and its suitability of the determination of this analyte in spiked water samples was studied. The pABs-based assay showed I50 = 17.95 ppb, I90 = 203.40 ppb, and I10 = 0.066 ppb, whereas the mABs-based assay showed I50 = 5.99 ppb, I90 = 485.40 ppb, and I10 = 0.074 ppb. The recoveries of fipronil from tap water samples by pABs-based ELISA were 93.00-124.00% in the range of 0-500 ng/mL, and those obtained from the samples by mABs-based ELISA were 94.70-108.00%. Different types of water from pool, river, and sea were spiked at different levels (ranging form 0.1 to 10 microg/L) and were assayed by the indirect ELISA with mABs. The recoveries of fipronil by this ELISA were in the range of 80-120%. The results demonstrate that this assay is suitable for the quantitative detection of fipronil at trace levels in water samples.     

Application of a monoclonal antibody-based enzyme-linked immunosorbent assay for the determination of ractopamine in incurred samples from food animals

A monoclonal antibody-based ractopamine immunoassay has been applied to incurred samples from sheep and cattle. Results obtained by immunoassay were compared with those from high-performance liquid chromatography (HPLC). Three sets of sample extracts containing primarily unmetabolized ractopamine were analyzed. Correlation of HPLC with enzyme-linked immunosorbent assay (ELISA) for beef liver samples gave an r(2) = 0.98 despite rather low ractopamine concentrations (range 1.1-13.4 ng/mL, n = 6). Ractopamine concentrations in cow urine samples treated by solid phase extraction, to remove ractopamine metabolites, also showed a high correlation between the HPLC and the ELISA results (r(2) = 0.95, range 1.0-275 ng/mL, n = 61). In contrast, HPLC and ELISA analyses of ractopamine in sheep urine were not well-correlated (r(2) = 0.58, range 0.85-51 ng/mL, n = 34). When ractopamine conjugates in urine samples were hydrolyzed with hydrolytic enzymes, ELISA and HPLC methods were highly correlated [r(2) = 0.94 for sheep (range 123-10 554 ppb, n = 60) and an r(2) = 0.98 for cattle (range 14-8159 ppb, n = 62)]. Tissues contained only minute amounts of ractopamine, and after 7-day withdrawal periods, less than 1 ppb of free ractopamine was detected. Ractopamine was rapidly metabolized in both cattle and sheep. The difference in ractopamine concentration of urine samples before and after hydrolysis indicated that only 1-5% of ractopamine was excreted unmetabolized. Results from this study indicate that the monoclonal antibody-based ELISA could be useful for a sensitive, quantitative, or qualitative ractopamine screening assay.     

Development of an enzyme-linked immunosorbent assay for the detection of pentachloronitrobenzene residues in environmental samples

A competitive enzyme-linked immunosorbent assay (ELISA) for pentachloronitrobenzene (PCNB), a fungicide and chemical intermediate, was developed using a polyclonal antiserum produced against a hapten-protein conjugate of pentachlorophenoxypropionic acid-bovine serum albumin (BSA). An indirect competitive ELISA of PCNB showed an IC50 of 37 ng/mL and a limit of detection (LOD) of 7 ng/mL. The ELISA can tolerate up to 10% (v/v) methanol, 5% (v/v) acetonitrile, or 5% (v/v) acetone without significant fluctuation of Amax and IC50. The assay sensitivity showed little change in a range of pH from 6 to 8 and concentrations of 0.05-0.2 M NaCl in the assay buffer. Very low cross-reactivities were observed for some structurally related compounds except for hexachlorobenzene (12%). The average recoveries of PCNB from fortified well water, river water, and soil samples were in ranges of 88-94, 80-91, and 70-81%, respectively. The correlations between the gas chromatographic and ELISA results were excellent (r 2 >or= 0.97, slopes from 0.86 to 1.10) for those fortified samples. The ELISA is a good alternative tool for monitoring PCNB residues in environmental samples.     

Development of two enzyme-linked immunosorbent assays for detection of endosulfan residues in agricultural products

Two competitive immunoassays, a laboratory assay based on microwell plates and a field test based on the use of polystyrene tubes, have been developed for the detection of endosulfan in agricultural products. The limit of detection for the microwell plate format was 0.8 +/- 0.1 microg/kg, and the limit of detection for the tube format was 1.6 +/- 0.2 microg/kg. A simple, rapid, and efficient extraction method was employed, and 76-112% recoveries of spiked samples were obtained. Methanol extracts of some agricultural product samples such as grape, carrot, spinach, and tobacco could be analyzed directly by immunoassay after dilution in 0.5% fish skin gelatin-phosphate buffered saline. In contrast, extracts of green tea caused significant interference in the assay, and a number of simple cleanup methods were ineffective in removing interference. However, use of the coagulating reagent polyvinyl pyrrolidone removed the matrix effect effectively. For the validation of the enzyme-linked immunosorbent assay (ELISA) tests, samples were analyzed by ELISA and gas chromatography (GC) after solid phase extraction. The relationship between data obtained using the tube assay and microwell assay was good (the lowest r(2) value was 0.94), and also, the immunoassay assay data correlated well with data obtained from GC analysis (the lowest r(2) value was 0.93). The developed immunoassay methods are the suitable methods for the rapid quantitative and reliable determination of endosulfan residues in agricultural products.     

Direct competitive enzyme-linked immunosorbent assay for sulfamethazine residues in milk

A direct competitive enzyme-linked immunosorbent assay (ELISA) is described for the detection and estimation of sulfamethazine residues in milk. Samples are cleaned up rapidly by acidifying and centrifuging the milk, adjusting the supernatant liquid to pH 7.0, and centrifuging again. The supernate is then assayed using set points to estimate sulfamethazine levels in the sample in the range of 1 ppb to 1 ppm. Multiple samples of milk can be screened in 1.5-2 h by this ELISA method.     

How good is a reconnaissance soil map for agronomic purposes?

Abstract. Information about the soil fertility status in irrigated ricelands at regional scales (1:50 000–1:250 000) is commonly not contained in classical soil maps. To assess the agronomic suitability of two different reconnaissance soil maps, we conducted a detailed soil survey in the Nueva Ecija province, Philippines. Soil samples were collected from 384 farmers' fields, and soil properties were measured for topsoil and subsoil samples. For most soil properties, a soil map made in 1940 (1:125 000) had within-map unit variances that were smaller than the total variance, whereas a new soil map of 1992 (1:50 000) did not significantly reduce the within-class variance. In both soil maps, classification into mapping units accounted for 0–40% of the variance of 14 agronomically important soil properties and large within-map unit variabilities were found. Underlying strategies of classical soil survey supported the partition of variance for relatively stable soil properties, such as soil texture, CEC, and organic matter. If reconnaissance soil maps are used in quantitative land evaluation studies, existing maps require upgrading by adding quantitative information about relevant soil properties and their within-map unit variability The sampling demand for upgrading a reconnaissance soil map was large, but pedotransfer functions can be used as cost-saving tools. Measures of soil nutrient status were highly variable within all mapping units and differences among farmers were much greater than the differences between soil types. Therefore, nutrient management in the study region should be based on individual field or farm recommendations rather than on soil-map based recommendations.     

Selenite fixation by soil particle-size separates

The fixation of selenite by clay- (< 2 μm), silt- (2–20 μm) and sand-size (20–2000 μm) separates from two arable soils was examined in solutions of ⁷⁵Se-labelled sodium selenite using a Se/sample ratio of 1/10⁶. Size separates were isolated by ultrasonic dispersion and gravity sedimentation. Selenite fixation was determined after equilibration periods ranging from 5 min to 26 h. Hydrogen peroxide-treated samples were included to examine the effect of organic matter on selenite fixation capacity.
The relative distribution of native Se, C, dithionite/citrate-extractable Fe and Al between size separates was similar. Concentrations in clay were four to nine times higher than in whole soils, silt showing two to five times higher concentrations and sand being very low in Se, C, Fe and Al.
After 1 h, clay, silt and sand fixed 64–65%, 45–61% and <5% of the selenite added, respectively. The fixation on whole soils was 14–18%. After 1 day, fixation on clay, silt, sand and whole soil increased to 78–87%, 67–79%, 3–14% and 31–39%, respectively.
Hydrogen peroxide-treatment reduced the selenite fixation capacity of whole soil, silt and sand to very low levels. Fixation on peroxide-treated clay was in accord with values for pure clay minerals reported in the literature. Generally, the fixation capacity of peroxide-treated natural clay and pure clay minerals was only half that observed for intact clay-size separates, demonstrating the importance of organic matter in soil selenite fixation capacity.     

Development of a multianalyte enzyme-linked immunosorbent assay for permethrin and aroclors and its implementation for analysis of soil/sediment and house dust extracts

Development of a multianalyte enzyme-linked immunosorbent assay (ELISA) for detection of permethrin and aroclors 1248 or 1254 and implementation of the assay for analysis of soil/sediment samples are described. The feasibility of using the multianalyte ELISA to monitor aroclors 1254 and permethrin simultaneously was tested with permethrin and aroclor standards and with aroclor- and permethrin-containing soil/sediment and house dust samples. Comparison of the I?? and I?? values of the multianalyte with those of a single-analyte assay revealed similar results, and multianalyte ELISA determination of analyte amounts in soil/sediment dust samples yielded similar results to those of a single-analyte assay. A single-analyte assay of permethrin content in permethrin-containing dust samples showed that the ELISA can determine the analyte accurately in samples with dust matrix contents ranging from 6.25 to 100 mg as indicated by the good correlation between the results of the immunoassay and those of the gas chromatography analysis.     

Enzyme-linked immunosorbent assay based on a polyclonal antibody for the detection of the insecticide fenitrothion. Evaluation of antiserum and application to the analysis of water samples.

For development of an indirect competitive enzyme-linked immunosorbent assay (ELISA) for the organophosphorus insecticide fenitrothion, the specificity of the antiserum R-3 generated with the bifunctional hapten, LysMNPA (2-[[[(3-methyl-4-nitrophenyl)oxy]methylcarbonyl]amino]-6-(2,4-dinitrophenyl)aminohexanoic acid) and the application to the residual analysis of some water samples were evaluated. At optimized ELISA conditions, the quantitative working range was from 1 to 39 ng/mL with a limit of detection of 0.3 ng/mL and an IC(50) value of 6 ng/mL. Cross-reactivity to structurally similar organophosphorus compounds and related chemicals was determined. The antiserum R-3 showed significant cross-reactivity with fenitrooxon and 3-methyl-4-nitrophenol, which have a 3-methyl-4-nitrophenoxy group as common structures, but showed relatively low cross-reactivity with other compounds. Each water sample (river water, tap water, purified water, and bottled water) had a matrix effect and was investigated by adding Tween 20 in the assay buffer. These four kinds of water samples were fortified with fenitrothion at several concentration levels and were directly analyzed with only dilution with an equal volume of antiserum solution. The mean recovery was 105.9%, and the mean coefficient of variation was 10.9%. The results suggested that the developed ELISA would be very suitable for a preliminary screening for fenitrothion in water samples at such low levels.     

Enzyme-linked immunosorbent assay (ELISA) and sol-gel-based immunoaffinity purification (IAP) of the pyrethroid bioallethrin in food and environmental samples

Enzyme-linked immunosorbent assay (ELISA) and sol-gel-based immunoaffinity purification (IAP) methods for the pyrethroid bioallethrin were developed and applied for monitoring bioallethrin in spiked food, soil, and dust samples. Attempts to determine bioallethrin content in fruit and vegetable extracts revealed high variability between sample preparations and marked interferences with the assay. Sol-gel IAP followed by solid-phase sample concentration was effective in removing the interfering components and resulted in high recovery of bioallethrin from spiked crude acetonic extracts of fruits and vegetables, even in the presence of high extract concentrations (28%). Solid-phase treatment alone failed to remove the interfering components from the spiked sample. Gas chromatography-mass spectrometry analysis of the IAP samples revealed bioallethrin as a doublet unsolved peak because of the cis and trans isomer present in the standard with confirmation of its mass. Unlike fruit and vegetable extracts, soil and dust samples did not interfere with the ELISA, and the bioallethrin content in those samples could be determined with high precision without the need of any further purification.     

Determination of the persistence of tetracycline antibiotics and their degradates in manure-amended soil using enzyme-linked immunosorbent assay and liquid chromatography-mass spectrometry

The persistence of manure-borne oxytetracycline in soil was investigated under field conditions. Soil cores were collected approximately once a month for over a period of two years and subsampled at depth increments of 0-5, 5-10, 10-15, 15-36, and 36-71 cm. Soil samples were analyzed by enzyme-linked immunosorbent assay (ELISA) and/or by liquid chromatography-mass spectrometry (LC-MS). Whereas LC-MS showed that oxytetracycline declined to <50% of its initial soil concentration after 3 weeks, ELISA showed that the total tetracyclines did not decline significantly 5 months after manure application. The differences between ELISA and LC-MS results are attributed to the broad cross-reactivity of the antibodies employed, which detect many structurally related tetracyclines, including their isomers and degradation products. Only trace amounts (< or = 1.0 microg/kg) of oxytetracycline were observed in the subsurface soil, and none was detected in water samples from field lysimeters, suggesting that oxytetracycline has low mobility in soil.     

Development of an immunoassay for the residues of the herbicide bensulfuron-methyl

To develop a competitive indirect enzyme-linked immunosorbent assay based on polyclonal antibodies for the detection of the sulfonylurea herbicide bensulfuron-methyl, seven structurally related haptens were synthesized. Four of them mimicking the target analyte were conjugated to keyhole limpet hemocyanin by the N-hydroxysuccinimide activated ester method to use as immunogens, and all of them were conjugated to bovine serum albumin to use as plate-coating antigens. Polyclonal antibodies raised in rabbits and the coating antigens were screened and selected for the assay in simple homologous and heterologous ELISA formats. Three sensitive heterologous ELISAs were selected and optimized, showing the average IC(50) values of bensulfuron-methyl as low as 0.17, 0.09, and 0.09 ng/mL, the detection ranges of 0.04-0.60, 0.01-0.60, and 0.04-0.25 ng/mL, and the lowest detection limits of 0.03, 0.002, and 0.03 ng/mL, respectively. The cross-reactivities of other sulfonylurea herbicides and metabolites of bensulfuron-methyl to the antibodies were less than 15% in the two assays. Recoveries from the analyte-fortified water samples in assay I were in the range of 81-125% by simple dilution. The correlation between the ELISA and HPLC was 0.999 (n = 15) with a slope of 1.37 in the analysis of groundwater samples fortified with bensulfuron-methyl. The results obtained strongly indicate that the ELISA can be a highly sensitive and convenient tool for detecting bensulfuron-methyl residues in agricultural and environmental samples.     

Hapten synthesis and development of polyclonal antibody-based multi-sulfonamide immunoassays

This paper reports the synthesis of five sulfonamide derivatives, the production of broad-specificity polyclonal antibodies for immunoassay of sulfonamides, and the analysis of milk samples by developed assay. The three-step synthesis procedure reported in most of the literature was adopted and modified in this study. In the procedure, the purification of the intermediate was avoided and the time of synthesis was shortened from >20 to 6-9 h with improved yields. This method is generally applicable to the synthesis of haptens containing the common structure of sulfonamides. Three haptens were coupled to keyhole limpet hemocyanin, and polyclonal antibodies were obtained from rabbits immunized with these conjugates. Using the antibodies obtained, from one of these was developed an enzyme-linked immunosorbent assay (ELISA) based on the competition between free sulfonamides and the hapten-horseradish peroxidase (HRP) conjugates. The hapten-HRP conjugate giving the best competitive results and 11 structurally different sulfonamides showed 50% inhibition at concentrations of <100 ng mL(-1). After removal of the protein with acetone, milk samples were analyzed by ELISA directly; a matrix effect could be avoided when a 1:20 dilution with phosphate-buffered saline was used, and 104-131% recoveries of spiked samples were obtained. The developed immunoassay is suitable to determine sulfisozole, sulfathiazole, sulfameter, sulfamethoxypyridazine, sulfapyridine, and sulfamethizole below the maximum residue limit in milk (100 ng mL(-1) of total sulfonamides) rapidly and reliably.     

Preparation of antibodies and development of an enzyme-linked immunosorbent assay (ELISA) for the determination of doxycycline antibiotic in milk samples

This paper reports the development of an immunoassay for the specific analysis of doxycycline (DC), a congener of the tetracycline antibiotic family (TCs), in milk samples. This is the first time that DC antibody production is reported, based on a rationally designed and well-characterized immunizing hapten. The chemical structure of the immunizing hapten (13-[(2-carboxyethyl)thiol]-5-hydroxy-6-α-deoxytetracycline, TC1) was designed to maximize recognition of the tetracycline characteristic moiety defined as lower periphery of the TCs plus the region of the upper periphery composed by the hydroxyl group at position C(5) (B ring) and the dimethylamino group in ring A. Polyclonal antibodies raised against TC1 coupled to horseshoe crab hemocianyn (HCH) were used to develop a homologous indirect competitive enzyme-linked immunosorbent assay (ELISA). The microplate ELISA can detect DC in buffer down to 0.1 μg L(-1). The ELISA has been proven to tolerate a wide range of ionic strengths and pH values. The assay is very selective for DC with a minor recognition of methacycline (32% of cross-reactivity). Experiments performed with whole milk samples demonstrate that samples can be directly analyzed after a simple treatment method, reaching detectability values below 5 μg L(-1).     

Development of monoclonal ELISAs for azinphos-methyl. 2. Assay optimization and water sample analysis.

Two enzyme-linked immunosorbent assays (ELISA) for the insecticide azinphos-methyl have been optimized and characterized. Both ELISAs are based on monoclonal antibodies produced from an immunogen with a hapten containing a phthalimido moiety and on protein conjugates of heterologous ligands containing a 1,2,3-benzotriazine group. Assay I was performed in the conjugate-coated ELISA format and assay II in the antibody-coated format. Several physicochemical factors (ionic strength, pH, incubation times, and Tween 20 and BSA concentrations) that influence assay performance were studied and optimized. Regarding specificity, both monoclonal immunoassays highly cross-reacted with azinphos-ethyl and phosmet. Finally, both assays were applied to the analysis of azinphos-methyl in spiked real water samples. For assay I the sensitivity, estimated as the I(50) value, was 0.40 nM, with a practical working range between 0.10 and 1.75 ng/mL and a limit of detection of 0.05 ng/mL. For assay II the sensitivity was 1.01 nM, with a practical working range between 0.32 and 2.54 ng/mL and a limit of detection of 0.08 ng/mL.