Determination of glyphosate residues in Egyptian soil samples.

A sensitive linker-assisted enzyme-linked immunosorbent assay (L'ELISA) was developed for the analysis of glyphosate in Egyptian soil samples. Polyclonal glyphosate antibodies were produced from rabbits immunized with glyphosate protein conjugate. The conjugate was prepared by activating the carboxylic groups of proteins; thyroglobulin or bovine serum albumin with 1-ethyl-3- (3-diaminopropyl) carbodiimide hydrochloride and N-hydroxysulfosuccinimide followed by directly coupled to the amino group of glyphosate. The L'ELISA used succinic anhydride to derivatize glyphosate, which mimics the epitopic attachment of glyphosate to thyroglobulin. L'ELISA recognized the derivatized glyphosate with a limit of detection (LOD) of 0.8 ng g-1 and sensitivity (IC50 value) of 0.018 µg g-1. The recovery values of the spiked soil samples with different concentrations of glyphosate were in the range of 87.4-97.2%. Good correlation was achieved between L'ELISA and conventional high-pressure liquid chromatography (HPLC) with fluorescence detection. This study demonstrated the utility and convenience of the sensitive, simple, practical and cost-effective L'ELISA method for glyphosate analysis in soil samples. Also, it is ideal for rapid screening of a large number of environmental samples.

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 IC50 value) 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.

Testicular deficiency associated with exposure to cypermethrin, imidacloprid, and chlorpyrifos in adult rats.

The testicular deficiency associated with exposure to three widely used insecticides in Egyptian agriculture was evaluated. Animals were orally treated with sub-lethal dose (1/50 of the oral LD50) of cypermethrin (CYP), imidacloprid (IMC), and chlorpyrifos (CPF) at 5, 9 and 1.9 mg/kg/day, respectively, five times a week for one month. The CYP, IMC, and CPF exposure resulted in a significant decline in animal body weight, sperm count, motility, normality, and viability with increased head and tail deformities. Significant reduction in serum testosterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), testis superoxide dismutase (SOD), and reduced glutathione (GSH) levels. In contrast, catalase (CAT), lipid peroxidation (LPO), and protein carbonyl content (PCC) levels were significantly stimulated. Jointly, obtained results were confirmed by microscopic examination of testis sections. The present data concluded that the CYP, IMC, and CPF have a public health impact and violently interferes with male rat reproductive system.

Development and validation of a novel enzyme-linked immunosorbent assay for monitoring ethylene thiourea in soil and vegetable samples.

A novel enzyme-linked immunosorbent assay (ELISA) was developed for the detection of ethylene thiourea (ETU); a metabolite of ethylenebisdithiocarbamates (EBDCs) fungicides such as mancozeb. The ELISA performed well under optimal conditions based one limit of detection (0.201 ng mL-1), sensitivity (IC50 3.71 µg mL-1), recovery (81.6-102.7%) and relative standard deviation (<12.5%). This method was used to determine ETU residues in soil and vegetables after 0, 1, 3, 5, 7 and 10 days following mancozeb application. Concentrations of ETU gradually increased in the soil up to the tenth day of application. While in tomato and potato samples, the maximum concentrations of ETU residues were reached on the third and fifth days, respectively, and then dissipated with half-lives of 1.33 and 4.33 days, respectively. The method was simple, rapid, cost-effective, environmentally friendly, and has the potential for high sample throughput.