Detailed cytotoxicity assessment of the formulated herbicide roundup classic and its constituents

Cytotoxicity of the globally market-leading herbicide ROUNDUP CLASSIC formulation and its components such as the active ingredient glyphosate and the formulating agent POEA (a mixture of polyethoxylated tallow amines) were investigated on the murine neuroectodermal stem cell-like (NE-4C) and osteoblastic (MC3T3-E1) cell lines. The cytotoxic and genotoxic effects on cell viability and cell cycles were evaluated based on the results of flow cytometry, enzymatic-assays, and alkaline single cell gel electrophoresis (Comet) assays, furthermore, the effects on cell morphology and dynamic mass redistribution of cellular contents were assessed with the use of the label-free Epic BenchTop optical biosensor on MC3T3-E1 cells adhered on the surface of the biosensor. Differences in the sensitivity of the investigated cell lines were detected, while the MC3T3-E1 cell line indicated less sensitivity to the effects of the treatments. Furthermore, differences were also observed in the sensitivity of the performed assays. The order of the inhibitory potency of the investigated compounds was as follows: glyphosate IPA salt << ROUNDUP CLASSIC < POEA. The applied Epic technique provides an effective tool for the real-time detection of cytotoxicity

In vivo and in vitro tests for the detection of biochemical and ecotoxicological effects of the herbicide active ingredient glyphosate

Aquatic organisms are outstandingly exposed to water contaminants because of their unavoidable contact with xenobiotics, thus their exposure needs to be routinely monitored. Due to its extensive use, the herbicidal agrochemical active ingredient glyphosate realizes massive exposure, its toxic effects alone and in formulations were evaluated in different in vivo aquatic ecotoxicological tests on various algae species, freshwater biofilm communities, Daphnia magna, and Danio rerio, furthermore the possible cytotoxic, genotoxic, and hormonemodulating effects were evaluated in vitro on different cell lines and test organisms. Significant differences were detected in the individual and combined toxicity of glyphosate and its coformulants presented in the formulations, therefore various additives cannot be classified as unequivocally inactive components. The result of the in vivo testing proved higher toxicity for the formulating agent and the formulation compared to the individual effect of glyphosate, and significant differences in the sensitivity of test species, and the effects on the sexual development of fish were also observed. The performed in vitro assays on cell lines demonstrated the potential cytotoxic and genotoxic effects of glyphosate and its formulations, and some of the effects are the result of the individual toxicity of glyphosate

Pesticide residues in environmental and produce samples from ecological and conventional paprika cultivation fields

To support environmental and food safety of spice paprika production, paprika growing sites in intensive and ecological cultivation have been sampled and analyzed for pesticide residues in Hungary. Two sites of three producers in each cultivation mode were sampled in early summer. Soil samples have been collected at three different points from two or three depth levels, thus, altogether 42 soil samples have been collected at six intensive cultivation fields (ICFs) and 23 soil samples from ecological cultivation fields (ECFs). Pesticide residues in soil extracts have been determined by gas chromatography coupled with mass spectrometry (GC-MS). In soils from ICF sampling sites pesticide active ingredients trifluralin, tefluthrin, chlorpyrifos and DDT were detected together with certain decomposition products (DDE, DDD). Harvested paprika samples were collected in September from four ICFs and from one ECF. Biological samples, prepared by a modified QuEChERS extraction method and analyzed for pesticide residues by GC-MS, contained no detectable amounts of pesticide active ingredients and metabolites, even when plants were grown in ICF on soil containing pesticide residues

Quality Management in Spice Paprika Production: From Cultivation to End Product

There is ample historical and scientifically proven information regarding the health benefits of spice paprika, including favourable physiological effects, anti-oxidant and anti-inflammatory properties. Nonetheless, even though it is consumed in small portions, spice paprika has occasionally been reported for chemical/microbiological contamination, as well as fraud or food adulteration. Quality management can guarantee effective reduction of such contamination cases. Different production stages within cultivation and production are subject to different contamination types. Cultivation is a common source of pesticide residues, and unfavourable harvest conditions may give rise to mycotoxins by pathogenic fungi. Storage and post-ripening prior to processing is attributed with microbial contamination and possible increase in mycotoxin content may significantly affect quality features. Technology steps, for example, washing, separation, drying may worsen microbial contamination or quality, but normally do not lead to increase in mycotoxins; nonetheless, decontamination technology is a prerequisite for microbial safety of the product. Upon effective decontamination, finishing steps in the processing technology, for example, grinding, packaging and end product handling do not affect the microbial status, but other, occasionally deliberate contamination due to mixing and adulteration may occur at this stage

Study on Soil Mobility of Two Neonicotinoid Insecticides

Movement of two neonicotinoid insecticide active ingredients, clothianidin (CLO) and thiamethoxam (TMX), was investigated in different soil types (sand, clay, or loam) and in pumice. Elution profiles were determined to explore differences in binding capacity. Soil characterized by high organic matter content retained the ingredients, whereas high clay content resulted in long release of compounds. Decrease in concentration was strongly influenced by soil types: both CLO and TMX were retained in loam and clay soils and showed ready elution through sandy soil and pumice. Elution capability of the active ingredients in sandy soil correlated with their water solubility, indicating approximately 30% higher rapidity for TMX than for CLO. Soil organic carbon-water partitioning coefficients (Koc) determined were in good agreement with literature values with somewhat lower value for CLO in sandy soil and substantially higher values for TMX in clay soil. High mobility of these neonicotinoid active ingredients in given soil types urges stronger precautionary approach taken during their application

A newly identified specific biological activity of glyphosate - inhibition of RGD-binding integrins

In this study we investigated the inhibitory effect of the widely used broad-spectrum herbicide active ingredient glyphosate and its related analogues on αVβ3 integrin binding to the shortest oligopeptide recognizing motif of integrins, the arginine-glycine-aspartic acid (RGD) sequence. Integrin binding characteristics were assessed in a modified enzyme linked immunosorbent assay (ELISA) and by a label-free optical biosensor technique. At 22 mM, glyphosate reached full inhibition of αVβ3, and the inhibitory activity of its main metabolite, aminomethylphosphonic acid (AMPA) was also above 95%, while another environmentally relevant metabolite, sarcosine exerted only a weaker effect, approximately 35% inhibition. In turn, the half maximal inhibitory concentration (IC50) of glyphosate and AMPA were reported to be 2.7±0.5 mM and 1.3±0.2 mM, respectively. The inhibitory effects of the other related compounds investigated (acetylglycine, glycine and iminodiacetic acid) at the same concentration, 22 mM were below 50%. Inhibitory effects on cell adhesion to RGD-modified surfaces by whole cells containing several types of RGD-binding integrins including αVβ3 were detected using the biosensor technique, where the integrin antagonist activity of glyphosate was also demonstrated

Chlorophyll fluorescence instrumentation for a rapid, in situ measurement of algal density

In the project reported, we are developing an instrument for measuring algal density based on the detection of chlorophyll fluorescence. Following the adjustment of several parameters defined during preliminary analyses, measurements were made on different concentrations of model green and blue algal cultures. Fluorescent signal intensities measured by the prototypes of the fluorometer module were compared to values determined by other, widely used methods for estimation of algal density (i.e. Bürker chamber cell counting, optical density measurement and chlorophyll-a measurement with ethanol extraction method). Fluorometer results correlated well with the other methods, resulting high correlation coefficients (R2>0.9%). Limits of detection and limits of quantification showed a decreasing trend during the development phases resulting in a highly sensitive instrument

Environmental monitoring of glyphosate and assessment of its combined cytotoxicity with adjuvants

Pesticide toxicology currently focuses mainly on two areas: long-term effects of given compounds and cocktail effects of chemicals, including combined effects of pesticide active ingredients with their adjuvants, as seen in the case of glyphosate-based herbicides. In this study surface water pollution in an agricultural region of Hungary by glyphosate was determined by ELISA method, and cytotoxic effects on HEK293 and NE-4C cells by glyphosate, its formulated herbicide (ROUNDUP® ) and adjuvant (polyethoxylated tallowamine, POEA) were compared. ROUNDUP and POEA were found to be equitoxic at short exposures (LC50: 10-15 ng/ml in 6 hrs), while glyphosate occurred to be of 500-750-fold less toxicity