Quantification of ionophores in aged poultry litter using liquid chromatography tandem mass spectrometry

Veterinary anticoccidials, biochemically known as ionophores, are widely used in poultry feed at therapeutic levels to treat Coccidiosis and at sub-therapeutic levels for growth- promotion. Commonly used ionophores in the US poultry industry are monensin, salinomycin, lasalocid and narasin. There is an increasing concern regarding the persistence of these anticoccidials in the environment. However, little attention has been directed to methods development for quantitatively measuring ionophores in complex environmental matrices such as poultry litters that are land applied. Here, we describe a rapid and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS)-based method developed for simultaneous quantification of monensin, lasalocid, salinomycin, and narasin in aged poultry litter samples. Results show significant level of monensin (97.8 ± 3.2 μg kg^?1), lasalocid (19.2 ± 6.6 μg kg^?1), salinomycin (70 ± 2.7 μg kg^?1) and narasin (57.3 ± 2.6 μg kg^?1) in poultry litter stored for over three years at < 5°C. Our findings indicate that even after several years of unmanaged storage of poultry litter, ionophores may continue to persist in this matrix, raising the possibility of prolonged release into the environment.     

Quantification of ionophores in aged poultry litter using liquid chromatography tandem mass spectrometry

Veterinary anticoccidials, biochemically known as ionophores, are widely used in poultry feed at therapeutic levels to treat Coccidiosis and at sub-therapeutic levels for growth- promotion. Commonly used ionophores in the US poultry industry are monensin, salinomycin, lasalocid and narasin. There is an increasing concern regarding the persistence of these anticoccidials in the environment. However, little attention has been directed to methods development for quantitatively measuring ionophores in complex environmental matrices such as poultry litters that are land applied. Here, we describe a rapid and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS)-based method developed for simultaneous quantification of monensin, lasalocid, salinomycin, and narasin in aged poultry litter samples. Results show significant level of monensin (97.8 ± 3.2?μg kg(-1)), lasalocid (19.2 ± 6.6?μg kg(-1)), salinomycin (70 ± 2.7?μg kg(-1)) and narasin (57.3 ± 2.6?μg kg(-1)) in poultry litter stored for over three years at < 5°C. Our findings indicate that even after several years of unmanaged storage of poultry litter, ionophores may continue to persist in this matrix, raising the possibility of prolonged release into the environment.     

Detection and quantitative analysis of 21 veterinary drugs in river water using high-pressure liquid chromatography coupled to tandem mass spectrometry

Introduction

The use of veterinary drugs in food production focuses on the control and improvement of animal health. The disadvantage of this practice is that pharmaceuticals and their metabolites are released into the environment, finding their way to natural water systems and becoming a potential risk to non-target organism.

Methods

This paper reports the development and validation of a quantitative method, based on high-performance liquid chromatography coupled to tandem mass spectrometry, for the simultaneous analysis of 21 veterinary drugs, antimicrobials, corticosteroids, coccidiostats and antifungal agents, in surface water.

Results

The precision of the method was established by calculating the mean recoveries, which were in the range of 94?C101%. The developed method was employed to conduct the first monitoring study on the presence of veterinary drugs in the Galicia region, Northwest of Spain and was applied to 235 surface water samples. Eleven veterinary drugs were detected at concentrations from below the limit of quantification to 2,978.6?ng?L^?1. Limits of detection and quantification were in the range of 6.2 (betamethasone, cortisone, decoquinate, dexamethasone, maduramycin, monensin, narasin, salinomycin, sulfachloropyridazine, sulfamethoxypyridazine and trimethoprim) to 12.5?ng?L^?1 (for the rest of the selected drugs) and 12.5 (betamethasone, cortisone, decoquinate, dexamethasone, maduramycin, monensin, narasin, salinomycin, sulfachloropyridazine, sulfamethoxypyridazine and trimethoprim) to 25.0?ng?L^?1 (for the remaining pharmaceuticals), respectively.

Conclusion

Sulfonamides were the group most frequently found, which are widely used in veterinary medicine.     

Fate and antibacterial potency of anticoccidial drugs and their main abiotic degradation products

The antibacterial potency of eight anticoccidial drugs was tested in a soil bacteria bioassay (pour plate method), EC₅₀-values between 2.4 and 19.6 μM were obtained; however, one compound, nicarbazin exhibited an EC₅₀-value above the maximum tested concentration (21 μM, 9.1 mg L⁻¹). The potency of mixtures of two of the compounds, narasin and nicarbazin, was synergistic (more than additive) with 10-fold greater antibacterial potency of the mixture than can be explained by their individual EC₅₀-values. The influence of pH, temperature, oxygen concentration and light on the transformation of robenidine and salinomycin was investigated. Robenidine was transformed by photolysis (DT₅₀ of 4.1 days) and was unstable at low pH (DT₅₀ of approximately 4 days); salinomycin was merely transformed at low pH, the latter into an unknown number of products. The antibacterial potency of the mixtures of transformation products of robenidine after photolysis and at low pH was comparable with that of the parent compound. Finally five photo-transformation products of robenidine were structural elucidated by accurate mass measurements, i-FIT values (isotopic pattern fit) and MS/MS fragmentation patterns.     

Determination of the insecticide diflubenzuron in mushrooms by kinetic method and high-performance liquid chromatographic method

In the present study, a new sensitive and simple kinetic-spectrophotometric method for the determination of the insecticide diflubenzuron [1-(4-chlorophenyl)-3-(2,6-diflubenzoil)urea] is proposed. The method is based on the inhibited effect of diflubenzuron on the oxidation of sulphanilic acid (SA) by hydrogen peroxide in phosphate buffer in presence Cu(II) ion. Diflubenzuron was determined with linear calibration graph in the interval from 0.31 to 3.1 μg mL^?1 and from 3.1 to 31.0 μg mL^?1. The optimized conditions yielded a theoretical detection limit of 0.18 μg mL^?1corresponding to 0.036 mg Kg^?1mushroom sample based on the 3S_b criterion. The RSD is 5.03–1.83 % and 2.81–0.71 % for the concentration interval of diflubenzuron 0.31–3.1 μg mL^?1and 3.1–31.0 μg mL^?1, respectively. The reaction was followed spectrophotometrically at 370 nm. The kinetic parameters of the reaction are reported, and the rate equations are suggested. The developed procedure was successfully applied to the rapid determination of diflubenzuron in spiked mushroom samples of different mushroom species. The HPLC method was used like a comparative method to verify results.     

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.     

Development and validation of LC-MS/MS method for the determination of Ochratoxin A and its metabolite Ochratoxin α in poultry tissues and eggs

The objective of this study was to develop a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the determination of Ochratoxin A (OTA) and Ochratoxin α (OTα) in poultry tissues and eggs. The two toxins were extracted by a mixture of acetonitrile/water, purified with a reversed phase C18 solid phase extraction column (SPE) and determined by LC-MS/MS. The LC-MS/MS method performances were evaluated in terms of linearity in solvent and in matrix (ranged from 0.5 to 15.10 µg L^?1 for OTA and from 0.60 to 17.85 µg L^?1 for OTα), limit of detection (LOD), limit of quantitation (LOQ), specificity, accuracy and precision in repeatability conditions. Recovery experiments were performed by spiking poultry liver, kidney, muscle and eggs around 1 µg kg^?1 and 10 µg kg^?1. LODs were 0.27 and 0.26 µg kg^?1 while LOQs were fixed at 1.0 and 1.2 µg kg^?1 for OTA and OTα, respectively. Main recoveries for OTA ranged from 82 to 109% and for OTα ranged from 55 to 89%. The values of within-laboratory relative standard deviation (RSD_r) were equal to or below 20%. Considering the results obtained and that all analytical performance criteria were fulfilled, the new extraction and purification method developed for OTA and OTα determination in animal tissues and eggs was found appropriate for control laboratories and research activities designed to ensure food safety.     

Background levels of polychlorinated biphenyls in British soils

The concentrations of PCBs in soil samples collected at rural and urban locations in Britain are reported. PCBs were detected in all samples analysed, within the range 2.3 to 444 μg kg⁻¹ (ppb), with a mean concentration of 22.8 μg kg⁻¹ and a median value of 7.2 μg kg⁻¹. A distinct sub-group of samples has a range of 2.3 to 19.2 μg kg⁻¹, with a mean of 7.5 μg kg⁻¹ and a median value of 6.8 μg kg⁻¹. These values are consistent with published data from other industrialised countries. Measurement was carried out using a two stage clean-up of soil extracts, with concentrated sulphuric acid and florisil, followed by gas chromatography and electron capture detection.     

Headspace solid phase microextraction method for determination of triazine and organophosphorus pesticides in soil

A headspace solid phase microextraction method (HS-SPME) for simultaneous determination of five pesticides belonging to triazine and organophosphorus pesticide groups in soil samples was developed. Microextraction conditions, such as temperature, extraction time and sodium chloride (NaCl) content were investigated and optimized using 100 μ m polydimethyl-siloxane (PDMS) fiber. Detection and quantification were done by gas chromatography-mass spectrometry (GC-MS). Relative standard deviation (RSD) and recovery values for multiple analysis of soil samples fortified at 30 μ g kg^{? 1} of each pesticide were below 13 % and higher than 70 %, respectively. Limits of detection (LOD) for all the compounds studied were less than 3.2 μ g kg^{? 1}. The proposed method was applied in the analysis of some agricultural soil samples.     

Ammonia volatilization loss from surface applied livestock manure

Ammonia (NH₃) emission from livestock manures used in agriculture reduces N uptake by crops and negatively impacts air quality. This laboratory study was conducted to evaluate NH₃emission from different livestock manures applied to two soils: Candler fins sand (CFS; light-textured soil, pH 6.8 and field capacity soil water content of 70 g kg^{? 1}) from Lake Alfred, Florida and Ogeechee loamy sand (OLS; medium-textured soil, pH 5.2 and field capacity soil water content of 140 g kg^{? 1}) from Savannah, Georgia. Poultry litter (PL) collected from a poultry farm near Douglas, Georgia, and fresh solid separate of swine manure (SM) collected from a farm near Clinton, North Carolina were used. Each of the soil was weighed in 100 g sub samples and amended with either PL or SM at rates equivalent to either 0, 2.24, 5.60, 11.20, or 22.40 Mg ha^{? 1} in 1L Mason jars and incubated in the laboratory at field capacity soil water content for 19 days to monitor NH₃ volatilization. Results indicated a greater NH₃ loss from soils amended with SM compared to that with PL. The cumulative NH₃volatilization loss over 19 days ranged from 4 to 27% and 14 to 32% of total N applied as PL and SM, respectively. Volatilization of NH₃ was greater from light-textured CFS than that from medium-textured OLS. Volatilization loss increased with increasing rates of manure application. Ammonia volatilization was lower at night time than that during the day time. Differences in major factors such as soil water content, temperature, soil type and live stock manure type influenced the diurnal variation in volatilization loss of NH₃ from soils. A significant portion (> 50%) of cumulative NH₃ emission over 19 d occurred during the first 5–7 d following the application of livestock manures. Results of this study demonstrate that application of low rates of livestock manure (≤ 5.60 Mg ha^{? 1}) is recommended to minimize NH₃ emissions.     

Anaerobic digestion of livestock and poultry manures spiked with tetracycline antibiotics

Abstract

We investigated the anaerobic degradation of tetracycline antibiotics (tetracycline [TC], oxytetracycline [OTC] and chlortetracycline [CTC]) in swine, cattle, and poultry manures. The manures were anaerobically digested inside polyvinyl chloride batch reactors for 64?days at room temperature. The degradation rate constants and half-lives of the parent tetracyclines were determined following first-order kinetics. For CTC the fastest degradation rate was observed in swine manure (k?=?0.016?±?0.001 d^?1; half-life = 42.8?days), while the slowest degradation rate was observed in poultry litter (k?=?0.0043?±?0.001 d^?1; half-life = 161?days). The half-lives of OTC ranged between 88.9 (cattle manure) and 99.0?days (poultry litter), while TC persisted the longest of the tetracycline antibiotics studied with half-lives ranging from 92.4?days (cattle manure) to 330?days (swine manure). In general, the tetracyclines were found to degrade faster in cattle manure, which had the lowest concentrations of organic matter and metals as compared to swine and poultry manures. Our results demonstrate that tetracycline antibiotics persist in the animal manure after anaerobic digestion, which can potentially lead to emergence and persistence of antibiotic resistant bacteria in the environment when anaerobic digestion byproducts are land applied for crop production.     

Short chain nitrocompounds as a treatment of layer hen manure and litter; effects on in vitro survivability of Salmonella,generic E. coli and nitrogen metabolism

The current study was conducted to assess the bactericidal effectiveness of several nitrocompounds against pathogens in layer hen manure and litter. Evidence from an initial study indicated that treatment of layer hen manure with 12 mM nitroethane decreased populations of generic E. coli and total coliforms by 0.7 and 2.2 log₁₀ colony forming units (CFU) g^?1, respectively, after 24 h aerobic incubation at ambient temperature when compared to untreated populations. Salmonella concentrations were unaffected by nitroethane in this study. In a follow-up experiment, treatment of 6-month-old layer hen litter (mixed with 0.4 mL water g^?1) with 44 mM 2-nitroethanol, 2-nitropropanol or ethyl nitroacetate decreased an inoculated Salmonella typhimurium strain from its initial concentration (3 log₁₀ CFU g^?1) by 0.7 to 1.7 log₁₀ CFU g^?1 after 6 h incubation at 37°C in covered containers. After 24 h incubation, populations of the inoculated S. Typhmiurium in litter treated with 44 mM 2-nitroethanol, 2-nitropropanol, ethyl nitroacetate or nitroethane were decreased more than 3.2 log₁₀ CFU g^?1 compared to populations in untreated control litter. Treatment of litter with 44 mM 2-nitroethanol, 2-nitropropanol, ethyl nitroacetate decreased rates of ammonia accumulation more than 70% compared to untreated controls (0.167 µmol mL^?1 h^?1) and loses of uric acid (< 1 µmol mL^?1) were observed only in litter treated with 44 mM 2-nitropropanol, indicating that some of these nitrocompounds may help prevent loss of nitrogen in treated litter. Results warrant further research to determine if these nitrocompounds can be developed into an environmentally sustainable and safe strategy to eliminate pathogens from poultry litter, while preserving its nitrogen content as a nutritionally valuable crude protein source for ruminants.     

Field Measurements of Natural and Fertilizer-Induced N2O Release Rates from Soils

The influence of the soil on atmospheric N₂O was studied by In-situ measurements in 1976–1979 at several field stations near Mainz, Germany, where different soil types were located. Measurements were carried out using the closed chamber method and applying stainless steel capillaries allowing soil air sampling down to 60 cm depth. The N2O In soil was found to be produced and consumed simultaneously In the uppermost soil layer resulting In a net flux of N₂O with release rates of 0.5–16 μg N₂O–Nm^?2h^?1 on unfertilized natural as well as agriculturally used soils. After fertilization with mineral fertilizers the N₂O release rates increased to values ≤43 μg N₂O–Nm^?2h^?1. The total amount of fertilizer-N released Into the atmosphere as N₂O was determined to be 0.01–0.05% for nitrate and 0.03–0.09 % for ammonium fertilizer.     

Fast extraction of polychlorinated dibenzo-p-dioxin and polychlorinated dibenzofuran in sewage sludge and soil samples

The current environmental legislations recommend monitoring chemical contaminants such as polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans before the use of sewage sludge on the agricultural land. In this study, a solid–liquid extraction with low-temperature purification (SLE-LTP) was optimized and validated to determine 2,3,7,8-tetrachlorodibenzo-p-dioxin and 2,3,7,8-tetrachlorodibenzofuran in sewage sludge and soil samples. The analyses were performed by gas chromatography-mass spectrometry operating in the selective ion mode (GC-MS-SIM). Acetonitrile:ethyl acetate 6.5:1.5 (v/v) was the best extraction phase, and the recoveries percentages were close to 100%. The linearity was demonstrated in the range of 1.25–25 µg L^?1 of 1.25–20 µg L^?1 for sewage sludge and soil, respectively. Matrix effect was proved for the two compounds and in the two matrices studied. Extraction percentages were between 78 and 109% and relative standard deviations ≤ 19%. The proposed method is faster than methods described in the literature because showed a few steps. The quantification limits (LOQ) in sewage sludge were 6.4 and 32 ng TEQ kg^?1 for 2,3,7,8-TCDF and 2,3,7,8-TCDD, respectively. In soil, LOQs were 0.8 and 8.0 ng TEQ kg^?1 for 2,3,7,8-TCDF and 2,3,7,8-TCDD, respectively. These values are lower than the maximum residue limits established by European Legislation. The method was applied to 22 agricultural soil samples from different Brazilian cities and 2,3,7,8-TCDF was detected in one of these samples.     

Dissipation and Leaching of 14C-Monocrotophos in Soil Columns under Subtropical Climate

Dissipation and leaching behavior of ¹⁴C-monocrotophos was studied for 365 days under field conditions using PVC cylinders. The first set (24 cylinders) was spiked with 1.0 μCi ¹⁴C-labeled monocrotophos along with 1.06 mg unlabeled monocrotophos to give a concentration of 2 mg kg ^?1 in the soil up to 15 cm depth. The second set (24 cylinders) received ¹⁴C-labeled monocrotophos along with other non-labeled insecticides viz., dimethoate @ 300 g a.i ha^?1, deltamethrin @ 12.5 g a.i ha^?1, endosulfan @ 750 g a.i ha^?1, cypermethrin @ 60 g a.i ha^?1, and triazophos @ 600 g a.i ha^?1 at an interval of 15 days each as recommended for the cotton crop. ¹⁴C-monocrotophos dissipated faster, up to 45% in first 90 days in columns treated with only monocrotophos compared to 25% in columns that received monocrotophos along with other insecticides. However, both the columns showed similar residues 180 days onward. After 180 days of treatment, 46% radiolabeled residues were observed, which reduced up to 39.6% after 365 days. Leaching of ¹⁴C-monocrotophos to 15–30 cm soil layer was observed in both the experimental setups. In the 15–30 cm soil layer of both soil columns, up to 0.19 mg ¹⁴C-monocrotophos kg^?1d. wt. soil was detected after 270 days.     

Emission factors and exposures from ground-level pyrotechnics

Potential exposures from ground-level pyrotechnics were assessed by air monitoring and developing emission factors. Total particulate matter, copper and SO₂ exposures exceeded occupational health guidelines at two outdoor performances using consumer pyrotechnics. Al, Ba, B, Bi, Mg, Sr, Zn, and aldehyde levels were elevated, but did not pose a health hazard based on occupational standards. Emission factors for total particulate matter, metals, inorganic ions, aldehydes, and polyaromatic hydrocarbons (PAHs) were determined for seven ground-supported pyrotechnics through air sampling in an airtight room after combustion. Particle generation ranged from 5 to 13% of the combusted mass. Emission factors (g Kg^?1) for metals common to pyrotechnics were also high: K, 23–45; Mg, 1–7; Cu, 0.05–7; and Ba, 0.03–6. Pb emission rates of 1.6 and 2.7% of the combusted mass for two devices were noteworthy. A high correlation (r² ≥ 0.89) between metal concentrations in pyrotechnic compositions and emission factors were noted for Pb, Cr, Mg, Sb, and Bi, whereas low correlations (r² ≤ 0.1) were observed for Ba, Sr, Fe, and Zn. This may be due to the inherent heterogeneity of multi-effect pyrotechnics. The generation of inorganic nitrogen in both the particulate (NO₂^?, NO₃^?) and gaseous (NO, NO₂) forms varied widely (<0.1–1000 mg Kg^?1). Aldehyde emission factors varied by two orders of magnitude even though the carbon source was carbohydrates and charcoal for all devices: formaldehyde (<7.0–82 mg Kg^?1), acetaldehyde (43–210 mg Kg^?1), and acrolein (1.9–12 mg Kg^?1). Formation of lower molecular weight PAHs such as naphthalene and acenaphthylene were favored, with their emission factors being comparable to that from the combustion of household refuse and agricultural debris. Ba, Sr, Cu, and Pb had emission factors that could produce exposures exceeding occupational exposure guidelines. Sb and unalloyed Mg, which are banned from consumer fireworks in the US, were present in significant amounts.     

Comparison of fatty acid profile in the chicken meat after feeding with narasin,nicarbazin and salinomycin sodium and phyto-additive substances

The purpose of this study was an experimental investigation and a statistical evaluation of the influence of various additives in feed mixtures of broiler chickens on fatty acids content and their ratio in breast and thigh muscles. First feed additive consisted of narasin, nicarbasin and salinomycin sodium, and other five additives were of phytogenic origin. In vivo experiment was realized on the poultry experimental station with deep litter breeding system. A total of 300 one-day-old hybrid chickens Cobb 500 divided into six groups were used for the experiment. The experimental period was divided into four phases, i.e. Starter, Grower 1, Grower 2 and Final, according to the application of commercial feed mixture of soy cereal type. Additive substances used in feed mixtures were different for each group. Basic feed mixtures were equal for all groups. Fatty acid profile of breast and thigh muscles was measured by the method of FT IR Nicolet 6700. Investigated additive substances in the feed mixtures did not have statistically significant effect on fatty acid content and omega-6/omega-3 polyunsaturated fatty acid (PUFA) ratio in breast and thigh muscles. Strong statistically significant relation between omega-6 PUFAs and total PUFAs were proved by experiment. A relation between omega-3 PUFAs and total PUFAs was found only in the group with Biocitro additive.     

Effect of phenobarbital and polychlorinated biphenyls on the toxicity and disposition of 1,2,4,5‐tetrachlorobenzene in the rat

Soil avoidance by earthworms has been generally considered a relevant and sensitive endpoint for assessing soil contamination by xenobiotics. However, when pesticide ecotoxicological assessment is concerned, the sensitivity of the recently standardized avoidance assay has been questioned. We hypothesized that this controversy may be due to the specific pesticide mode of action of the chemicals used rather than reveal inconsistencies in the test feasibility, i.e. provided that no pesticides interfering with neuronal pathways are tested, this bioassay should keep expected high levels of sensitivity. In this study, the avoidance behaviour of the earthworm Eisenia andrei under exposure to the carbamate insecticide methomyl [S-methyl N-(methylcarbamoyloxy)thioacetimidate] was linked to the corresponding acetylcholinesterase (AChE) inhibition. Significant AChE inhibition occurred at lower concentrations (from 0.86 mg Kg^?1 onwards) than significant avoidance of spiked soil (from 5.62 mg Kg^?1 onwards). This indicates that assessments regarding pesticides that have neurotoxic activity may be biased if behavioral endpoints are selected. Despite theoretical hypothesis that have been raised, this should be the first study providing preliminary experimental evidence on such a link between avoidance behavior and neuronal impairment levels in earthworms. Further studies are ongoing that should refine conclusions of this study.     

PCB in the U.K. environment — A preliminary survey

Data on PCB levels in soil and grass samples from rural and urban locations in the United Kingdom are presented, on a dry weight basis. Average PCB concentrations for rural areas are 8 μg kg⁻¹ in soil (range 1 to 23 μg kg⁻¹) and 9 μg kg⁻¹ in grass (range 7 to 16 μg kg⁻¹); for urban areas, 43 μg kg⁻¹ in soil (range 11 to 141 μg kg⁻¹) and 40 μg kg⁻¹ in grass (one value); for industrial locations, 41 μg kg⁻¹ in soil (range 20 to 67 μg kg⁻¹). The levels fall within the range of values reported for other countries.     

Preparation of antibodies and development of an enzyme immunoassay for determination of atrazine in environmental samples

An indirect competitive enzyme-linked immunosorbent assay (ELISA) has been developed and optimized for atrazine determination in soil at different depths (0–10, 10–20, and 20–30 cm) before and after 48 h of application, corn shoot and cow milk samples collected from Dina farm, Egypt. This assay was based on a specific polyclonal antibodies (PAb) raised by immunizing New Zealand rabbits with an immunogen prepared by coupling 3-{4-(ethylamino)-6-(isopropylamino)-1,3,5-triazine-2-yl} thiopropanoic acid to bovine serum albumin (BSA) via N-hydroxysuccinimide (NHS) active ester method. The sensitivity (estimated as IC₅₀value) was 17.5 μg mL^?1 with a detection limit of 0.1 ng mL^?1. The maximum atrazine concentration was found in soil especially in the deepest layer (325 and 890 μg kg^?1 before and after application, respectively). Atrazine concentration in corn shoot was 333.28, μg kg^?1 dry plant, while there was no detectable amount in milk. All samples screened by ELISA were validated by gas chromatography mass spectrometer procedure (GC/MS). Good correlation was achieved between the two methods (r = 0.997 for soil and 0.9814 for plant). This study demonstrates the utility and convenience of the simple, practical and cost–effective ELISA method in the laboratory for analysis of environmental samples. The method is ideal for the rapid screening of large numbers of samples in laboratories where access to GC/MS facilities, is limited or lacking.