2-cyano-n-((ethylamino)carbonyl)-2-(methoxyimino)acetamide and famoxadone

As a new protective and therapeutic fungicide, studies on famoxadone-cymoxanil are rare, and its toxicity to aquatic organisms has not been reported. In the present study, zabrafish embryos were exposed to several concentrations of famoxadone-cymoxanil at 10 hpf. Then, the changes of their shape, heart rate, development and function of innate and adaptive immune cells, oxidative stress, apoptosis, the expression of apoptosis-related genes and immune-related genes, the locomotor behavior were observed and detected in acute toxicity of famoxadone-cymoxanil. Our studies showed that, after exposure to famoxadone-cymoxanil, zebrafish embryos had decreased heart rate, shortened body length, swollen yolk sac. Secondly, the number of innate and adaptive immune cells was significantly reduced; and neutrophil migration and retention at the injury area were inhibited, indicating the developmental toxicity and immunotoxicity of famoxadone-cymoxanil on the zebrafish. We also found that the oxidative stress related indicators of embryos were changed significantly, and apoptosis were substantially increased. Further investigation of changes of some key genes in TLR signaling including TLR4, MYD88 and NF-κB p65 revealed that the mRNA expression of these genes was up-regulated. Meanwhile, the mRNA expression of some proinflammatory cytokines such as TNF-α, IFN-γ, IL6 and IL-1β was also up-regulated. In addition, the activity, the total distance, time and average speed were decreased along with the increase of exposure concentration. The absolute turn angle, sinuosity and the enzymatic activity of acetylcholinesterase (AChE) were also increased. These results suggested that famoxadone-cymoxanil can induce developmental toxicity, immunotoxicity and neurobehavioral toxicity in zebrafish larvae.

Topics: Acetamides; Animals; Apoptosis; Behavior, Animal; Embryo, Nonmammalian; Embryonic Development; Fungicides, Industrial; Motor Activity; Oxidative Stress; Strobilurins; T-Lymphocytes; Toll-Like Receptor 4; Zebrafish

Famoxadone-cymoxanil is a new protective and therapeutic fungicide, but little research has been done on it or its toxicity in aquatic organisms. In this study, we used zebrafish to investigate the cardiotoxicity of famoxadone-cymoxanil and the potential mechanisms involved. Zebrafish embryos were exposed to different concentrations of famoxadone-cymoxanil until 72 h post-fertilization (hpf), then changes of heart morphology in zebrafish embryos were observed. We also detected the levels of oxidative stress, myocardial-cell proliferation and apoptosis, ATPase activity, and the expression of genes related to the cardiac development and calcium-signaling pathway. After famoxadone-cymoxanil exposure, pericardial edema, cardiac linearization, and reductions in the heart rate and cardiac output positively correlated with concentration. Although myocardial-cell apoptosis was not detected, proliferation of the cells was severely reduced and ATPase activity significantly decreased, resulting in a severe deficiency in heart function. In addition, indicators of oxidative stress changed significantly after exposure of the embryos to the fungicide. To better understand the possible molecular mechanisms of cardiovascular toxicity in zebrafish, we studied the transcriptional levels of cardiac development, calcium-signaling pathways, and genes associated with myocardial contractility. The mRNA expression levels of key genes in heart development were significantly down-regulated, while the expression of genes related to the calcium-signaling pathway (ATPase [atp2a1], cardiac troponin C [tnnc1a], and calcium channel [cacna1a]) was significantly inhibited. Expression of klf2a, a major endocardial flow-responsive gene, was also significantly inhibited. Mechanistically, famoxadone-cymoxanil toxicity might be due to the downregulation of genes associated with the calcium-signaling pathway and cardiac muscle contraction. Our results found that famoxadone-cymoxanil exposure causes cardiac developmental toxicity and severe energy deficiency in zebrafish.

Topics: Acetamides; Animals; Apoptosis; Calcium Signaling; Cardiotoxicity; Down-Regulation; Embryo, Nonmammalian; Fungicides, Industrial; Gene Expression Regulation, Developmental; Heart; Heart Rate; Kruppel-Like Transcription Factors; Oxidative Stress; Strobilurins; Water Pollutants, Chemical; Zebrafish; Zebrafish Proteins

A method was validated for the simultaneous analysis of the residues of dimethomorph, famoxadone and cymoxanil in grape and raisin matrix by ethyl acetate based extraction and liquid chromatography tandem mass spectrometric analysis. Field studies were conducted to evaluate the dissipation rate kinetics and processing factor (PF) for these pesticides during raisin making. Residue data during the drying process were best fitted to 1st+1st order rate kinetics with half-life ranging between 8-9 days for dimethomorph, 12-13 days for famoxadone and 9-10 days for cymoxanil at single dose (SD) and double dose (DD), respectively. PF values calculated were 1.03 and 1.14 for dimethomorph, 1.95 and 2.09 for famoxadone, and 1.99 and 1.35 for cymoxanil at SD and DD, respectively. PF value >1 indicates concentration of the residues during raisin making. The residues of detected pesticides in market samples of raisins were devoid of any risk of acute toxicity related to dietary exposure.

Topics: Acetamides; Chromatography, Liquid; Mass Spectrometry; Methacrylates; Morpholines; Oxazoles; Strobilurins; Vitis

The extensive use of plant protection products in urban and rural pest control obliges to develop valid analytical methods for their successful and reliable quality control. An analytical procedure for the simultaneous determination of famoxadone and cymoxanil, by reversed phase high performance liquid chromatography based on the use of a monolithic C18 column has been developed and validated. The method involved the extraction of the active ingredients by sonication of the sample with acetonitrile and direct injection on a reversed phase liquid chromatographic system. The repeatability of the method expressed as relative standard deviation (%RSD), was lower than 1 % for both compounds. The limits of quantification for famoxadone and cymoxanil were 10 and 16 µg mL(-1) respectively. The new method involves a considerable reduction in time for both sample preparation and analysis. The proposed analytical procedure is accurate and precise.

Topics: Acetamides; Chromatography, High Pressure Liquid; Environmental Monitoring; Methacrylates; Oxazoles; Pesticides; Strobilurins