Application of an enzyme-linked immunosorbent assay for the detection of clenbuterol residues in swine urine and feeds

The present study outlines applications of an enzyme-linked immunosorbent assay (ELISA) for the analysis of clenbuterol residues. Antisera were raised from rabbits immunized with diazotized clenbuterol-bovine serum albumin (BSA) conjugate. The assay was specific to clenbuterol with a half-maximum inhibition concentration (IC₅₀) of 1.8 ng/mL and 2.5 ng/mL in blank swine urine and phosphate buffer solution, respectively. The assay had high cross-reactivity (86%) with mabuterol, but low with other adrenergic agonists and antagonists. The average recovery of clenbuterol, as measured with the ELISA, ranged from 90% to 112% in swine urine samples and from 86% to 95% in feeds, respectively. This new assay was compared with commercial ELISA test kits. An excellent correlation (r ² = 0.98) between the two methods and satisfactory recoveries suggest that the new assay can be suitable for the determination of clenbuterol residues in real samples. The assay was used to analyze clenbuterol residues in 103 swine urine samples and 68 feed samples collected from northern China. Approximately 50% of the urine samples and 25% of the feed samples analyzed were found positive (concentration of clenbuterol ≥ 1 ppb). The results indicate that clenbuterol was misused in some of the areas surveyed.     

Fate in soil of 14C-sulfadiazine residues contained in the manure of young pigs treated with a veterinary antibiotic

The fate of ¹⁴C-labeled sulfadiazine (¹⁴C-SDZ) residues was studied in time-course experiments for 218 days of incubation using two soils (A_p horizon of loamy sand, orthic luvisol; A_p horizon of silt loam, cambisol) amended with fresh and aged (6 months) ¹⁴C-manure [40 g kg^?1 of soil; 6.36 mg of sulfadiazine (SDZ) equivalents per kg of soil], which was derived from two shoats treated with ¹⁴C-SDZ. Mineralization of ¹⁴C-SDZ residues was below 2% after 218 days depending little on soil type. Portions of extractable ¹⁴C (ethanol-water, 9:1, v/v) decreased with time to 4–13% after 218 days of incubation with fresh and aged ¹⁴C-manure and both soils. Non-extractable residues were the main route of the fate of the ¹⁴C-SDZ residues (above 90% of total recovered ¹⁴C after 218 days). These residues were high immediately after amendment depending on soil type and aging of the ¹⁴C-manure, and were stable and not remobilized throughout 218 days of incubation. Bioavailable portions (extraction using CaCl₂ solution) also decreased with increasing incubation period (5–7% after 218 days). Due to thin-layer chromatography (TLC), 500 μg of ¹⁴C-SDZ per kg soil were found in the ethanol-water extracts immediately after amendment with fresh ¹⁴C-manure, and about 50 μg kg^?1 after 218 days. Bioavailable ¹⁴C-SDZ portions present in the CaCl₂ extracts were about 350 μg kg^?1 with amendment. Higher concentrations were initially detected with aged ¹⁴C-manure (ethanol-water extracts: 1,920 μg kg^?1; CaCl₂ extracts: 1,020 μg kg^?1), probably due to release of ¹⁴C-SDZ from bound forms during storage. Consistent results were obtained by extraction of the ¹⁴C-manure-soil samples with ethyl acetate; portions of N-acetylated SDZ were additionally determined. All soluble ¹⁴C-SDZ residues contained in ¹⁴C-manure contributed to the formation of non-extractable residues; a tendency for persistence or accumulation was not observed. SDZ's non-extractable soil residues were associated with the soluble HCl, fulvic acids and humic acids fractions, and the insoluble humin fraction. The majority of the non-extractable residues appeared to be due to stable covalent binding to soil organic matter.     

Fate and availability of glyphosate and AMPA in agricultural soil

The fate of glyphosate and its degradation product aminomethylphosphonic acid (AMPA) was studied in soil. Labeled glyphosate was used to be able to distinguish the measured quantities of glyphosate and AMPA from the background values since the soil was sampled in a field where glyphosate had been used formerly. After addition of labeled glyphosate, the disappearance of glyphosate and the formation and disappearance of AMPA were monitored. The resulting curves were fitted according to a new EU guideline. The best fit of the glyphosate degradation data was obtained using a first-order multi compartment (FOMC) model. DT₅₀ values of 9 days (glyphosate) and 32 days (AMPA) indicated relatively rapid degradation. After an aging period of 6 months, the leaching risk of each residue was determined by treating the soil with pure water or a phosphate solution (pH 6), to simulate rain over a non-fertilized or fertilized field, respectively. Significantly larger (p < 0.05) amounts of aged glyphosate and AMPA were extracted from the soil when phosphate solution was used as an extraction agent, compared with pure water. This indicates that the risk of leaching of aged glyphosate and AMPA residues from soil is greater in fertilized soil. The blank soil, to which 252 g glyphosate/ha was applied 21 months before this study, contained 0.81 ng glyphosate/g dry soil and 10.46 ng AMPA/g dry soil at the start of the study. Blank soil samples were used as controls without glyphosate addition. After incubation of the blank soil samples for 6 months, a significantly larger amount of AMPA was extracted from the soil treated with phosphate solution than from that treated with pure water. To determine the degree of uptake of aged glyphosate residues by crops growing in the soil, ¹⁴C-labeled glyphosate was applied to soil 6.5 months prior to sowing rape and barley seeds. After 41 days, 0.006 ± 0.002% and 0.005 ± 0.001% of the applied radioactivity was measured in rape and barley, respectively.     

Simple Model of Dissolved Oxygen Consumption in a Bay within High Organic Loading: An Applied Remediation Tool

San Vicente Bay is a coastal shallow embayment in Central Chile with multiple uses, one of which is receiving wastewater from industrial fisheries, steel mill effluents, and domestic sewage. A simulation model was developed and applied to dissolved oxygen consumption by organic residues released into this embayment. Three compartments were established as function of: depth, circulation and outfall location. The model compartments had different volumes, and their oxygen saturation value was used as baseline. The parameters: (a) BOD₅ of the industrial and urban effluents, (b) oxygen demand by organic sediments, (c) respiration, (d) photosynthesis and (e) re-aeration were included in the model. Iteration results of the model showed severe alterations in Compartment 1, with a decrease of 65% in the oxygen below saturation. Compartment 2 showed a small decline (10%) and compartment 3 did not show apparent changes in oxygen values. Measures recommended for remediation were to decrease the BOD₅ loading by 30% in the affected sector. Iteration of the model for 200 h following recommendations derived from the preceding results produced an increase in saturation of 60% (5 ml O₂ L⁻¹), which suggested an improvement of the environmental conditions.     

Pesticide Residues in Bovine Milk from a Predominantly Agricultural State of Haryana, India

One hundred forty seven samples of bovine milk were collected from 14 districts of Haryana, India during December 1998–February 1999 and analysed for the presence of organochlorine pesticide (OCPs) residues. ∑HCH, ∑DDT, ∑endosulfan and aldrin were detected in 100%, 97%, 43% and 12% samples and with mean values of 0.0292, 0.0367, 0.0022 and 0.0036 μg/ml, respectively. Eight percent samples exceeded the maximum residue limit (MRL) of 0.10 mg/kg as recommended by WHO for ∑HCH, 4% samples of 0.05 mg/kg for α-HCH, 5% samples of 0.01 mg/kg for γ-HCH, 26% samples of 0.02 mg/kg for β-HCH as recommended by PFAA and 24% samples of 0.05 mg/kg as recommended by FAO for ∑DDT. Concentrations of β-HCH and p,p′-DDE were more as compared to other isomers and metabolites of HCH and DDT.     

Insecticide Contamination of Jamaican Environment. V. Island-Wide Rapid Survey of Residues in Surface and Ground Water

Residues of organochlorines and organophosphates were determined by gas chromatography in water and sediment from 26 locations in 17 major rivers, 7 natural springs and 13 wells across Jamaica. Samples were collected on only one occasion between May and July, 1994. Residues of endosulfan were detected in all but three rivers; -endosulfan in 15 samples of sediment (0.9–108.1, mean = 28.93, S.E. = 7.198 g kg-1) and 13 of water (0.01–0.35, mean = 0.11, S.E. = 0.035 g L-1), -endosulfan in 5 sediment (15.29–49.35, mean = 30.56, S.E. = 7.132 g kg-1) and 12 water (0.05–0.31, mean = 0.14, S.E. = 0.031 g L-1) samples, and endosulfan sulphate in waters of three rivers (0.003–0.244 g L-1). Chlorpyrifos was present in 9 sediment (0.423-135.2, mean = 18.38, S.E. = 10.699 g kg-1) and two water (0.001–0.022 g L-1) samples, diazinon and ethoprophos in the sediment of one river each. Mean levels (g L-1) of and isomers and sulphate of endosulfan were 0.16 (S.E. = 0.057), 0.12 (S.E. = 0.036) and 0.15 (S.E. = 0.089), respectively, in four of the seven springs and 0.23 (S.E. = 0.052), 0.11 (S.E. = 0.029) and 0.26 (S.E. = 0.088), respectively, in seven of the thirteen wells monitored.     

Experimental investigation of the efficiency of triple rinsing of agricultural containers regarding their characterization as non-hazardous wastes

The efficiency of water rinsing for removing fenthion from non-combustible used pesticide containers was investigated. Four hundred empty containers from four olive oil producing areas in Greece were studied. Following triple rinsing cleaning procedures, the amount of fenthion retained in the container ranged from 0.01% to 0.1% w/w of its initial concentration. Extraction was accomplished with acetone, and the extracts were analyzed with a capillary gas chromatographic system coupled with flame ionization detector. The results obtained showed that triple rinsing is an effective procedure for removing fenthion from used pesticide containers, since the remaining pesticide concentration was below the legislated threshold value. Consequently, the containers can be handled as non-hazardous solid wastes.     

Validation of QuEChERS-based method for determination of flusilazole residues in grape by high-performance liquid chromatography with photodiode array detector

A high-performance liquid chromatography with photodiode array detector analytical method was developed to determine the residue levels of flusilazole in grape and investigate the dissipation pattern and safety. The results showed that the mean recoveries were in the range of 86%–90%. Limits of detection and quantification were 0.008 and 0.02 mg/kg, respectively. The residue levels of flusilazole were best described by first-order rate kinetics and with half-lives ranging from 4.2 days (recommended dose) to 4.6 days (double the recommended dose). In conclusion, flusilazole at the recommended or even at double the recommended doses does not pose any apparent hazards to consumers.     

Extractable atrazine and its metabolites in agricultural soils from the temperate humid zone

Extractable atrazine and its metabolites (hydroxyatrazine, deethylatrazine and deisopropylatrazine) were evaluated in agricultural soils from the temperate humid zone (Galicia, NW Spain) under laboratory conditions. The experiment was performed with five soils with different properties (organic C, soil texture and atrazine application history), both unamended and treated with atrazine at field application rate. Measurements of the atrazine compounds were made at different time intervals (1, 3, 6, 9 and 12 weeks) during a 3-month incubation period. Results showed that only hydroxyatrazine was detected in the extractable fraction of the unamended soils, with values remaining relatively constant throughout the incubation period. Atrazine addition notably increased the concentration of the parent compound and its degradation products; deisopropylatrazine and hydroxyatrazine were the main metabolites detected in the extractable fraction of the treated soils, whereas deethylatrazine was not detected. After 7 days incubation, values of total extractable residues, expressed as percentage of initially added atrazine, ranged from 75 to 86% (25–68% of atrazine, 7–11% of hydroxyatrazine and 9–57% of deisopropylatrazine). The values decreased rapidly during the first 3 weeks of incubation, showing values of 2–8% in soils with higher atrazine application and from 28 to 30% in soils with lower application history. At the end of the incubation, 2–8% of total extractable residues were still detected (0–4% of atrazine, 2–3% of hydroxyatrazine and 0–2% of deisopropylatrazine), indicating a residual effect of atrazine addition. These variations in the extractable fraction indicated that most added atrazine was rapidly degraded, especially in soils with higher application history.     

活性炭辅助微波热解污泥反应条件的试验研究

针对微波不能直接实现原污水污泥高温热解的问题,采用活性炭作为微波能吸收物质辅助污泥热解。对影响污泥热解效能的3个主要因素：污泥样品量、活性炭掺杂量和微波辐射功率,各做了3个水平的考察,得到了试验条件下的最优方案：污泥样品量30g、活性炭掺杂量6g、微波辐射功率1200W。结果表明：在最优试验方案下,污泥能在7min内升至920℃的高温,实现污泥快速、高效热解。通过分析3个因素对污泥热解效能的影响,进一步对最优试验方案下的固体产物吸收微波的性能及用其作为掺杂物辅助微波热解污水污泥的可行性进行了研究,结果表明：固体残渣吸收微波性能良好,可以代替活性炭作为更经济、高效的污泥微波热解辅助材料。