Tebuconazole is a chiral trizole fungicide and widely used in many crops for controlling disease. Tebuconazole is potential toxic to some aquatic organisms but relative information of its isomers is scarce. To detect the endocrine disrupting effects and difference of rac-, R-, and S-tebuconazole, the chitinase activity in Daphnia magna and chitobiase activity in each test medium were used as biomonitors after a 14-day exposure. Results showed that chitinase activity was significantly reduced by rac-, R-, and S-tebuconazole. The chitobiase activity in the test medium was reduced by rac- and R-tebuconazole before day 10, and only one peak was observed at day 10 or day 12 compared with two obvious peaks in the control group (days6 and 12). S-tebuconazole delayed and reduced the reproduction of D. magna, but did not delay the first chitobiase activity peak, whereas the second peak could not be characterized as the exposure concentration and time increased. Compared with chitinase activity, chitobiase activity can still be used as a rudimentary model for identifying molt-interfering xenobiotics, and further studies should focus on the analysis of correlations between these parameters.
A sensitive and specific method for the determination of propineb and its metabolites, propylenethiourea (PTU) and propylenediamine (PDA), using gas chromatography with flame photometric detection (GC-FPD) and LC-MS/MS was developed and validated. Propineb and its metabolite residue dynamics in supervised field trials under Good Agricultural Practice (GAP) conditions in banana and soil were studied. Recovery of propineb (as CS2), PDA and PTU ranged from 75.3 to 115.4% with RSD (n = 5) of 1.3-11.1%. The limit of quantification (LOQ) of CS2, PDA and PTU ranged from 0.005 to 0.01mg kg(-1), and the limit of detection (LOD) ranged from 0.0015 to 0.0033mg kg(-1). Dissipation experiments showed that the half-life of propineb in banana and soil ranged from 4.4 to 13.3days. PTU was found in banana with a half-life of 31.5-69.3days, while levels of PDA were less than 0.01mg kg(-1) in banana and soil. It has been suggested that PTU is the major metabolite of propineb in banana. The method was demonstrated to be reliable and sensitive for the routine monitoring of propineb and its metabolites in banana and soil. It also serves as a reference for the detection and monitoring of dithiocarbamates (DTCs) residues and the evaluation of their metabolic pathway.
A simple, sensitive, reliable method was developed for the simultaneous determination of organochlorine and pyrethriod pesticide residues in Chinese patent medicines Six ingredient rehmannia pills and Xiaoyao pills. These pesticides were extracted by ethyl acetate. The extraction time and volume of ethyl acetate were optimized. Cleanup of extracts was performed with dispersive-solid phase extraction using graphitized carbon black as the sorbent. The determination of pesticides in the final extracts was carried out by gas chromatography-tandem mass spectrometry in multiple reaction monitoring mode (GC-MS/MS, MRM). The linearity of the calibration curves is good in matrix-matched standard and yields the coefficients of determination (R-2) 0.99 for all of the target analytes. Under optimized conditions, the average recoveries (five replicates) for most pesticides range from 75.5% to 114.6%, and RSDs are less than 10.0%. The LODs of 18 pesticides in Six ingredient rehmannia pill and Xiaoyao pills are in the range of 0.01-8.82g kg(-1). The developed method meets the requirements of pesticide residue analysis and could be effectively used for routine analysis of the organochlorine and pyrethriod pesticide residues in Six ingredient rehmannia pills and Xiaoyao pills.
The aim of this study was to investigate in vitro effects of fenitrothion (FNT) on mouse splenic lymphocytes. Here, naive mice had their spleens harvested and splenocytes isolated. After exposure to FNT for 48hr: splenocyte viability was measured using a tetrazolium dye assay; cell phenotypes, i.e., B-cells (CD19(+)), T-cells (CD3(+)), and T-cell subsets (CD4(+) and CD8(+)), were quantified by flow cytometry; and, production of cytokines/granzyme-B was assessed via enzyme-linked immunosorbent assay. The ability for FNT to induce oxidative stress in the cells was evaluated by measuring hydroxyl radical (<bold>OH</bold>) and malondialdehyde (MDA) production and changes in glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activity. The results showed that FNT significantly inhibited splenocyte proliferation, and decreased production of interleukin (IL)-2, interferon gamma, IL-4, and granzyme B, but had no impact on IL-6 production. FNT also selectively decreased splenic T-cell levels but did not induce changes in CD19(+) B-cells. Further, within the T-cell populations, percentages of CD3(+), CD4(+), and CD8(+) T-cells (particularly CD8(+) T-cells) were reduced. Lastly, FNT selectively increased MDA and <bold>OH production and inhibited SOD and GSH</bold>-Px activities in the splenic lymphocytes. These findings suggest that, due to oxidative damage, FNT selectively inhibits splenic T-lymphocyte survival and cytokine/granzyme production in vitro.
The degradation of bifenthrin (BF) and chlorpyrifos (CP), either together or individually, by a bacterial strain (CB2) isolated from activated sludge was investigated. Strain CB2 was identified as belonging to genus Pseudomonas based on the morphological, physiological, and biochemical characteristics and a homological analysis of the 16S rDNA sequence. Strain CB2 has the potential to degrade BF and CP, either individually or in a mixture. The optimum conditions for mixture degradation were as follows: OD600nm = 0.5; incubation temperature = 30 degrees C; pH = 7.0; BF-CP mixture (10mg L-1 of each). Under these optimal conditions, the degradation rate constants (and half-lives) were 0.4308d(-1) (1.61d) and 0.3377d(-1) (2.05d) for individual BF and CP samples, respectively, and 0.3463d(-1) (2.00d) and 0.2931d(-1) (2.36d) for the BF-CP mixture. Major metabolites of BF and CP were 2-methyl-3-biphenylyl methanol and 3,5,6-trichloro-2-pyridinol, respectively. No metabolite bioaccumulation was observed. The ability of CB2 to efficiently degrade BF and CP, particularly in a mixture, may be useful in bioremediation efforts.
To investigate the degradation activity of the manganese ABC transporter, vegetative catalase 1 and acetoin dehydrogenase E1 from Bacillus subtilis YB1, the proteins were prokaryotically expressed and purified. Assay results showed that the three enzymes were able to degrade nicosulfuron (2- (4,6-dimethoxypyrimidine-2-pyrimidinylcarbamoylaminosulfonyl) -N,N-dimethylnicotinamide), with vegetative catalase 1 exhibiting the highest activity. To further examine the degradation pathway, the degradation products of the three enzymes and the YB1 strain were detected by liquid chromatography-mass spectrometry(LC-MS). The nicosulfuron degradation products of the three enzymes were consistent with those of the YB1 strain, indicating the presence of two pathways: one due to cleavage of sulfonylurea bridges and ring-opening of 1-(4,6-dimethoxy-pyrimidin-2-yl)-3-(2-methyliminomethanesulfonyl-acetyl)ureaas the pyrimidine ring, yielding the product; and the another due to cleavage of a sulfonylurea bridge, yielding 4,6-dihydroxy pyrimidine (111 m/z), 2-ylamine -4,6-dimethoxy pyrimidine and ((4-(dimethycarbamoyl)pyridine-2-yl)sulfonyl)carbamic acid as products, which were further degraded to 4,6-dihydroxy pyrimidine and KN-dimethyl-2-sulfamoyl-isonicotinamide. The above results reveal a major contribution of extracellular enzymes to the degradation of nicosulfuron by the YB1 strain. Our data help in elucidation of the mechanism of nicosulfuron bio-degradation and may facilitate the construction of engineered strains.
This study aimed to determine the contents of main mineral elements of wild Boletus edulis and to assess its edible safety, which may provide scientific evidence for the utilization of this species. Fourteen mineral contents (Ba, Ca, Cd, Co, Cr, Cu, Fe, Mg, Mn, Na, Ni, Sr, V and Zn) in the caps and stipes of B. edulis as well as the corresponding surface soils collected from nine different geographic regions in Yunnan Province, southwest China were determined. The analyses were performed using inductively coupled plasma atomic emission spectrometer (ICP-AES) after microwave digestion. Measurement data were analyzed using variance and Pearson correlation analysis. Edible safety was evaluated according to the provisionally tolerable weekly intake (PTWI) of heavy metals recommended by United Nations Food and Agriculture Organization and World Health Organization (FAO/WHO). Mineral contents were significantly different with the variance of collection areas. B. edulis showed relative abundant contents of Ca, Fe, Mg and Na, followed by Ba, Cr, Cu, Mn and Zn, and the elements with the lower content less were Cd, Co, Ni, Sr and V. The elements accumulation differed significantly in caps and stipes. Among them, Cd and Zn were bioconcentrated (BCF > 1) while others were bioexcluded (BCF < 1). The mineral contents in B. edulis and its surface soil were positively related, indicating that the elements accumulation level was related to soil background. In addition, from the perspective of food safety, if an adult (60 kg) eats 300 g fresh B. edulis per week, the intake of Cd in most of tested mushrooms were lower than PTWI value whereas the Cd intakes in some other samples were higher than this standard. The results indicated that the main mineral contents in B. edulis were significantly different with respect to geographical distribution, and the Cd intake in a few of regions was higher than the acceptable intakes with a potential risk.
The rapid development and increase of antibiotic resistance are global phenomena resulting from the extensive use of antibiotics in human clinics and animal feeding operations. Antibiotics can promote the occurrence of antibiotic resistance genes (ARGs), which can be transferred horizontally to humans and animals through water and the food chain. In this study, the presence and abundance of ARGs in livestock waste was monitored by quantitative PCR. A diverse set of bacteria and tetracycline resistance genes encoding ribosomal protection proteins (RPPs) from three livestock farms and a river were analyzed through denaturing gradient gel electrophoresis (DGGE). The abundance of sul(I) was 10(3) to 10(5) orders of magnitude higher than that of sul(II). Among 11 tet-ARGs, the most abundant was tet(O). The results regarding bacterial diversity indicated that the presence of antibiotics might have an evident impact on bacterial diversity at every site, particularly at the investigated swine producer. The effect of livestock waste on the bacterial diversity of soil was stronger than that of water. Furthermore, a sequencing analysis showed that tet(M) exhibited two genotypes, while the other RPPs-encoding genes exhibited at least three genotypes. This study showed that various ARGs and RPPs-encoding genes are particularly widespread among livestock.
Fungicide pyraclostrobin has been widely employed to control plant diseases by inhibiting the mitochondrial respiration of pathogenic fungi. Due to its broad spectrum, the extensive use of pyraclorstrobin was reported to cause emerging resistance on crops. Here, we evaluated the control effect of 250g L-1 of pyrachlostrobin suspension concentrate (SC) against freckle disease (caused by Phyllosticta spp) on banana. Meanwhile, the dissipation and residue dynamics of pyraclostrobin in banana and soil under field conditions were determined by high performance liquid chromatography (HPLC) with DAD detection in different locations. The analytical method was validated using spiked samples at three levels, which indicated the recoveries ranged from 92.0% to 99.0% with relative standard deviations (RSDs) below 5%, providing a sensitive, precise and reliable method to monitor pyraclostrobin in banana fruit and soil. The dissipation of pyraclostrobine followed the first-order kinetics and its half-lives were 5.25 to 9.90days. In addition, the terminal residues of pyraclostrobin in banana, banana sarcocarp and soil were below the maximum residue limit (MRL) (0.02mg kg(-1)) after a pre-harvest interval (PHI) of 42days, which suggesting that the use of pyraclostrobin at recommended dosages was safe to banana and the environment. In summary, we demonstrated the integrated evaluation on the disease control capacity of pyraclostrobin and its environmental behavior on banana, aiming to provide solid and basic data for the safe use of fungicide pyraclostrobin.
In attempt to evaluate the effects of cyclodextrins (CDs) on enantioselectivity of chiral pesticides toxicity, this study investigated effects of three kinds of cyclodextrins including -CD, -CD and randomly methylated -CD (RAMEB) on toxicity of four enantiomers of isomalathion including (1R, 3R)-isomalathion, (1S, 3S)-isomalathion, (1S, 3R)-isomalathion and (1R, 3S)-isomalathion. Generally, the addition of -CD and RAMEB (1.5g/L to 3.5g/L concentration) could lead to reduction of isomalathion toxicity in most cases, while the presence of -CD (0.3g/L to 1.5g/L concentration) enhanced the toxicity of isomalathion. It was speculated that higher electronic cloud density and lower water solubility of -CD than -CD and RAMEB might favor to combination between acetylcholinesterase (AChE) and isomalathion included by -CD. With respect for -CD and RAMEB, isomalathion included by them could be easily dissolved in water because of high water solubility of the two CDs. Therefore, -CD and RAMEB can be used as remediation regent for the pollution of isomalathion, and -CD can act as an additive in improving bioactivity of such pesticides. In addition, the presence of CDs can alter enantioselectivity of chiral pesticides. The differences on the extent of enantioselectivity variation of isomalathion induced by -CD, RAMEB and -CD might be ascribe to the different cavity, electron cloud density and solubility among the three CDs. In conclusion, the above results gave researchers a possibility to change enantioselectivity of chiral pesticides from undesirable outcomes to desirable ones.
A rapid and quantitative method for the determination of N-6-Benzylademine (N-6-BA) was established through the application of surface-enhanced Raman spectroscopy (SERS). The Raman peak intensities of N-6-BA at 1002cm(-1) positively correlated to N-6-BA concentrations in sprout extracts. The R-2 reached 0.99, and RSDs calculated below 10% at the concentration range of 0.1 approximate to 5g mL(-1). The average recoveries were 80.0% approximate to 98.2% for blank samples intentionally contaminated at differing levels of 0.04, 0.4, and 1g g(-1). The whole procedure, including sample preparation and SERS detection, did not exceed 30min for a set of 6 samples. This study indicates that SERS is a promising technique for rapid tracing analysis and on-site testing of N-6-BA.
The degradation dynamics and terminal residues of dufulin enantiomers were investigated in two typical corn plants. A convenient and precise chiral method by high-performance liquid chromatography coupled with tandem triple quadrupole mass spectrometry (HPLC/MS/MS) using a Chiralpak IC column was developed and validated for measuring dufulin enantiomers in corn plants and corn. The two enantiomers of dufulin quickly dissipated in the corn plant, and no noticeable stereoselectivity was observed during degradation or in the final residues. After 30% rac-dufulin wettable powder with a 1- to 1.5-fold dose of the recommended value was sprayed two to three times on corn plants, the residue levels of S-(+)-dufulin and R-(-)-dufulin in corn from both sites were lower than or equal to 0.0520 mg kg(-1) on days 7, 14 and 21 after the last application. The dietary risk assessment indicated that dufulin did not exhibit obvious dietary health risks in corn samples when good agricultural practices were implemented. The findings from this study may be used to better understand the chiral profiles of dufulin in the environment and the effect of dufulin residues in corn on health.
A simultaneous method for quantifying eight metabolites of organophosphate pesticides and pyrethroid pesticides in urine samples has been established. The analytes were extracted using liquid-liquid extraction coupled with WCX solid phase extraction (SPE) cartridges. Eight metabolites were chemically derivatized before analysis using gas chromatography-tandem mass spectrometry (GC-MS-MS). The separation was performed on a HP-5MS capillary column (30m x 0.25mm x 0.25 mu m) with temperature programming. The detection was performed under electro-spray ionization (ESI) in multiple reaction monitoring (MRM) mode. An internal standard method was used. The extraction solvent, types of SPE cartridges and eluents were optimized by comparing the sample recoveries under different conditions. The results showed that the calibration curves of the five organophosphorus pesticides metabolites were linear in the range of 0.2-200g/L (r(2) 0.992) and that of the three pyrethroid pesticides metabolites were linear in the range of 0.025-250g/L (r(2) 0.991). The limits of detection (LODs, S/N 3) and the limits of quantification (LOQs, S/N 10) of the eight metabolites were 0.008-0.833g/L and 0.25-2.5g/L, respectively. The recoveries of the eight metabolites ranged from 54.08% to 82.49%. This efficient, stable, and cost-effective method is adequate to handle the large number of samples required for surveying the exposure level of organophosphorus and pyrethroid pesticides in the general population.
The goal of the present study was to elucidate the in vitro cytotoxicity of Roundup and to reveal the possible related mechanisms in L-02 hepatocytes. By detecting reactive oxygen species (ROS) production, glutathione (GSH)/superoxide dismutase (SOD) levels, mitochondrial permeability transition pore (PTP) open rate, apoptosis-inducing factor (AIF) release, intracellular Ca2+ concentration, and alanine aminotransferease (ALT)/aspartate aminotransferase (AST) leakage, we determined that Roundup induced anti-oxidant system inhibition, mitochondria damage, DNA damage, membrane integrity and permeability changes, and apoptosis in L-02 hepatocytes. By revealing the mechanistic insights of Roundup-induced cytotoxicity, our results are valuable for the design of preventive and therapeutic strategies for the occupational population exposed to Roundup and other pesticides.
Trichlorfon is an organophosphate insecticide that is widely used in aquaculture and agriculture against parasitic infestations and has caused aquatic toxicity to non-target organisms. To evaluate the effects of low doses of trichlorfon on the oxidative stress and hepatotoxicity in amphibians, Chinese brown frogs (Rana chensinensis) were exposed to trichlorfon at concentrations of 0, 0.01, 0.1, and 1.0mg/L for 2 and 4weeks. Then, the activities of superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), and the content of malondialdehyde (MDA) in hepatic tissue were examined to evaluate the effects of oxidative stress and lipid peroxidation. The histopathological alternations to the liver were observed through light and transmission electron microscopy (TEM). The results showed that SOD and CAT activities were increased in the livers of frogs exposed to various concentrations of trichlorfon. The GST activity showed no significant changes at any concentration after 2weeks of exposure, whereas there was an initial increase after exposure to 0.1mg/L of trichlorfon at 4weeks. The content of MDA revealed a significant decrease after exposure. Histopathological and ultrastructural studies showed that trichlorfon induced hyalinization, vacuolation, nucleus necrosis, and cellular swelling in hepatocytes. These results suggest that low doses of trichlorfon could induce oxidative stress, lipid peroxidation, and hepatic lesions in frogs, which shows that even lower, non-lethal doses of trichlorfon are potentially toxic to amphibians.