Development of a sensitive,competitive, indirect ELISA for the detection of fumonisin B1 in corn originating from Anhui province,China

Fumonisin B₁ (FB₁) is a secondary metabolite produced by Fusarium verticillioides or Fusarium proliferatum, which present in food and feed. It causes hazardous effects on human and animal health. A monoclonal antibody (mAb) against FB₁ was produced and a simple, reliable and sensitive, competitive, indirect enzyme-linked immunosorbent assay (ci-ELISA) for detection of FB₁ was developed and the experiment conditions were optimized. The coating concentration of FB₁-ovalbumin (FB₁-OVA) was 500 ng mL^?1, the action concentrations of anti-FB₁ mAb and goat anti-mouse IgG were 1.28 × 10⁴ and 1:5000, respectively. The 50% inhibitory concentration (IC₅₀) was 11 ng mL^?1, with a detectable range of 1.25–250 ng mL^?1, and a limit of determination (LOD) of 1.15 ng mL^?1. The cross-reactivity (CR) of the antibody against fumonisin B₂ (FB₂) was 60.4, and <1% against deoxynivalenol (DON), aflatoxin B₁ (AFB₁), ochratoxin A (OTA) or zearalenone (ZEN). In spiked samples (250 ng g^?1, 500 ng g^?1, 1000 ng g^?1), the mean recoveries ranged from 86.7 ± 5% to 102 ± 4%, and the coefficient of variation (CV) ranged from 3% to 10%. A survey of 96 corn samples from Bozhou, Fuyang, Bengbu, and Hefei, in Anhui province, China, was performed. Frequencies of FB₁ contamination were 83.3%, 95.8%, 20.8% and 91.7%, and the mean concentrations of positive samples were 0.702 μg kg^?1, 0.883 μg kg^?1, 0.074 μg kg^?1, and 0.276 μg kg^?1, respectively. The results of this study suggest that the ci-ELISA developed in this study can be used to identify FB₁ in corn, furthermore, further study is needed to investigate FB₁ contamination in food and feed to prevent its harmful health effects.     

A new sensitive test based on yeast cells for studying environmental pollution

Different tests based on yeast cells were developed for determination of mutagenic/carcinogenic action; however, they all showed lower sensitivity compared to bacterial tests, the main reason for this being the limited permeability of yeast cells. We found that general permeability of Saccharomyces cerevisiae cells can be increased by mutation and on this basis we developed a more sensitive test. The aim of this study was to prove the applicability of our test, called D7ts1, in environmental studies. Soil, water and air samples were taken during 1998 from regions in Bulgaria with declared low, average or high pollution levels and investigated for presence of mutagenic/carcinogenic activities in the bacterial test of Ames, the yeast D7 test of Zimmermann and our new D7ts1 test. Results obtained evidenced the following conclusions: (1) the usage of D7ts1 test instead of D7 test permits a clearer measurement of positive samples and detects mutagenic/carcinogenic activities undetectable by D7 test; (2) all samples with positive Ames test were positive in the D7ts1 test; however, some samples, clearly positive in the D7ts1, were negative in the Ames test; therefore, the simultaneous usage of D7ts1 and Ames tests in environmental studies is advantageous because it detects dangers for the human health activities to which bacterial cells do not respond; and (3) regions in Bulgaria declared clean were found to be polluted; particularly troubled are the whole-year positive data in the three tests for air samples from a 'clean' region.     

A semi-target analytical method for quantification of OH-PCBs in environmental samples

Environmental Science and Pollution Research - Hydroxylated polychlorinated biphenyls (OH-PCBs) are oxidative metabolites of PCBs and residuals found in original Aroclors. OH-PCBs are known to play...     

A new rapid method for quantification of PCBs in transformer oil

In order to improve the efficiency and cost-effectiveness of the PCB analysis, the DMSO partition and SPE extraction were applied to clean up the PCB-contaminated transformer oils and PCB level was determined by means of the quadrupole GC/MS. The analysis data obtained from this method were compared statistically to that from the standard method.     

A modified UCT method for biological nutrient removal: configuration and performance

A pilot-scale prototype activated sludge system is presented, which combines both, the idea of University of Cape Town (UCT) concept and the step denitrification cascade for removal of carbon, nitrogen and phosphorus. The experimental set-up consists of an anaerobic selector and stepwise feeding in subsequent three identical pairs of anoxic and oxic tanks. Raw wastewater with influent flow rates ranging between 48 and 168 l d(-1) was fed to the unit at hydraulic residence times (HRTs) of 5-18 h and was distributed at percentages of 60/25/15%, 40/30/30% and 25/40/35% to the anaerobic selector, 2nd and 3rd anoxic tanks, respectively (influent flow distribution before the anaerobic selector). The results for the entire experimental period showed high removal efficiencies of organic matter of 89% as total chemical oxygen demand removal and 95% removal for biochemical oxygen demand, 90% removal of total Kjeldahl nitrogen and total nitrogen removal through denitrification of 73%, mean phosphorus removal of 67%, as well as excellent settleability. The highest removal efficiency and the optimum performance were recorded at an HRT of about 9h and influent flow rate of 96 l d(-1), in which 60% is distributed to the anaerobic selector, 25% to the second anoxic tank and 15% to the last anoxic tank. Consequently, the plant configuration enhanced removal efficiency, optimized performance, saved energy, formed good settling sludge and provided operational assurance.     

A fast and efficient modified sectional method for simulating multicomponent collisional kinetics

A fast and efficient method for simulating the evolution of internally mixed multicomponent particle size distributions for aerosol coagulation and droplet coalescence is developed. The technique is based upon a bin-wise sectionalization of the particle mass domain and by imposing the condition of mass conservation for each component. The distribution of each species as a function of the total particle mass is represented in each mass bin as a two-parameter exponential function. Particles of a given mass are assumed to be internally homogeneously mixed. The method is shown to be numerically stable for a wide range of time steps. The numerical solution is compared with both analytical results and results from other well-accepted numerical schemes. This comparison reveals that the proposed technique offers the advantage of being fast and accurate, even for coarse spectral resolution. The method is computationally attractive and easily allows the treatment of ten or more different chemical species in a collisionally evolving particle size distribution. The applicability of the method is demonstrated with several examples: Coalescence growth of multicomponent cloud droplet spectra, coagulation of measured multi-species aerosol particle size distributions, and the simulation of the accumulation mode due to a source of small aerosol particles. The technique is ideally suited for modelling the interaction of microphysics and chemistry in a size-bin resolving aerosol or cloud model.     

Production of monoclonal antibody and application in indirect competitive ELISA for detecting okadaic acid and dinophytoxin-1 in seafood

Background, aim, and scope

Okadaic acid (OA) and analogues of dinophysistoxin (DTX) are key diarrheic shellfish poisoning (DSP) toxins, which possibly arouse DSP symptoms by consuming the contaminated shellfish. Because of the stable toxicity in high temperature and the long-term carcinogenicity, the outbreaks of DSP related to consumption of bivalve mollusks contaminated by DSP toxins pose a hazard to public health. Therefore, it is worth developing a fast and reliable analytical method for the detection of OA and analogues in shellfish. In this paper, an indirect competitive enzyme-linked immunosorbent assay (ELISA) (icELISA) for detecting OA and DTX-1 in seafood was developed based on monoclonal antibody (McAb).

Methods

The OA was conjugated to human immunoglobulin G (IgG) and bovine serum albumin (BSA) by the active ester method as the immune antigen and the detective antigen. The spleen cells from BALB/c mice immunized with OA-IgG were fused with SP2/0 myeloma cells. A hybridoma cell line, which secreted McAb against OA, was selected by ??limiting dilution?? cloning. An icELISA was developed based on immobilized conjugate (OA-BSA) competing the McAb with the free OA in seafood sample.

Results

A hybridoma cell line, which secreted IgG1 subclass monoclonal antibody (McAb) against OA, was selected. The IC₅₀ of the McAb for OA and dinophytoxin-1 (DTX-1) were 4.40 and 3.89 ng/mL, respectively. Based on the McAb, an indirect competitive ELISA for detection of OA and DTX-1 in seafood was developed. The regression equation was y?=?54.713x???25.879 with a coefficient correlation of R ²?=?0.9729. The linear range and the limit of detection were 0.4?C12.5 and 0.45 ng/mL, respectively. The average recovery of OA and DTX-1 spiked shellfish was 82.29% with the coefficient of variation of 7.67%.

Conclusion

The developed icELISA is a fast, sensitive, and convenient assay for detecting of total amount of OA and DTX-1 in seafood.     

A practical method for quantification of phosphorus- and glycogen-accumulating organism populations in activated sludge systems.

Enhanced biological phosphorus removal (EBPR) from wastewater relies on the enrichment of activated sludge with phosphorus-accumulating organisms (PAOs). The presence and proliferation of glycogen-accumulating organisms (GAOs), which compete for substrate with PAOs, may be detrimental for EBPR systems, leading to deterioration and, in extreme cases, failure of the process. Therefore, from both process evaluation and modeling perspectives, the estimation of PAO and GAO populations in activated sludge systems is a relevant issue. A simple method for the quantification of PAO and GAO population fractions in activated sludge systems is presented in this paper. To develop such a method, the activity observed in anaerobic batch tests executed with different PAO/GAO ratios, by mixing highly enriched PAO and GAO cultures, was studied. Strong correlations between PAO/GAO population ratios and biomass activity were observed (R2 > 0.97). This served as a basis for the proposal of a simple and practical method to quantify the PAO and GAO populations in activated sludge systems, based on commonly measured and reliable analytical parameters (i.e., mixed liquor suspended solids, acetate, and orthophosphate) without requiring molecular techniques. This method relies on the estimation of the total active biomass population under anaerobic conditions (PAO plus GAO populations), by measuring the maximum acetate uptake rate in the presence of excess acetate. Later, the PAO and GAO populations present in the activated sludge system can be estimated, by taking into account the PAO/GAO ratio calculated on the basis of the anaerobic phosphorus release-to-acetate consumed ratio. The proposed method was evaluated using activated sludge from municipal wastewater treatment plants. The results from the quantification performed following the proposed method were compared with direct population estimations carried out with fluorescence in situ hybridization analysis (determining Candidatus Accumulibacter Phosphatis as PAO and Candidatus Competibacter Phosphatis as GAO). The method showed to be potentially suitable to estimate the PAO and GAO populations regarding the total PAO-GAO biomass. It could be used, not only to evaluate the performance of EBPR systems, but also in the calibration of potential activated sludge mathematical models, regarding the PAO-GAO coexistence.     

A rapid and sensitive fluoroimmunoassay based on quantum dot for the detection of chlorpyrifos residue in drinking water

A rapid and sensitive indirect competitive fluorescence-linked immunosorbent assay (cFLISA) method based on quantum dots as the fluorescence label coupled with secondary antibody (Ab₂) for the detection of chlorpyrifos in drinking water has been developed. The cFLISA method allowed for chlorpyrifos determination in a liner working range of 15.2–205.5 ng mL^?1. The 50 % inhibition value (IC₅₀) and the limit of detection (LOD) of the cFLISA were 50.2 ng mL^?1 and 8.4 ng mL^?1, while the IC₅₀ and the LOD of the conventhional enzyme linked immunosorbent assay (ELISA) were 95.3 ng- mL^?1 and 16.2 ng mL^?1, respectively. When the concentrations of chlorpyrifos were 200, 100 and 50 ng mL^?1, the recoveries ranged from 90.8 % to 108.2 % with a coefficient of variation (CV) of 7.5 %–15.2 %. In water sample analysis, the results of cFLISA were similar to those obtained from a cELISA and a high performance liquid chromatography (HPLC) method, while the detection time by cFLISA was reduced 0.5 h compared with ELISA. It showed that cFLISA could be used as a new screening method for the detection of pesticide residue.     

Bisphenol A and Bisphenol S release in milk under household conditions from baby bottles marketed in Italy

A simple and sensitive validated analytical method based on liquid chromatography coupled to tandem fluorescence (FD) and ultraviolet (UV) spectrophotometry was applied to monitor the presence of bisphenol A and bisphenol S in plastic baby bottles marketed in Italy. The limits of detection (LOD) were 3.75 ng mL^?1 and 80.00 ng mL^?1, and those of quantification (LOQ) were 12.51 ng mL^?1 and 260.00 ng mL^?1 for BPA (FD detection) and for BPS (UV detection), respectively. BPA was found in only four samples, two samples undergone to microwave heating and two samples undergone to bottle warmer heating either at 40°C or at 80°C. Although the quantities of leached BPA were well below the reference dose for daily intake established by the European Food Safety Authority (EFSA) (4.0 µg kg^?1 bw/day), the release of BPA and BPS from these plastic materials should be carefully considered by the government authorities to increase people's awareness on this issue and to protect the most vulnerable population group.     

Development of indirect competitive ELISA using egg yolk-derived immunoglobulin (IgY) for the detection of Gentamicin residues

Gentamicin (Gent) is an aminoglycoside antibiotic being used in livestock sector. Gent residues could cause some genetic disorders by nonsense mutations. This study aimed to develop IgY-based ELISA for the detection of Gent in animal products. Gent was conjugated with Bovine serum albumin (BSA) by carbodiimide method for further immunization in the laying chickens. PEG-6000 extraction method was employed to extract IgY from the egg yolk. The titer of anti-Gent-IgY attained the peak of 1:256,000 after the 5^th booster immunization. Checkerboard titration confirmed that, anti-Gent IgY in 1:2,000 dilution could give an Optical Density (OD) 1.0 at 2 µg mL^?1 of Gent-OVA coating concentration. IgY-based indirect competitive ELISA (Ic-ELISA) showed that, the IC₅₀ value of anti-Gent IgY was 2.69 ng mL^?1 and regression curve equation was y = ?16.27x + 56.97 (R² = 0.95, n = 3), confirming that, the detection limit (LOD, IC₁₀ value) was 0.01 ng mL^?1. Recoveries from fresh milk, pork and chicken samples were ranged from 69.82% to 94.32%, with relative standard deviation lower than 10.88%. Our results suggested that generated anti-Gent IgY antibodies can be used in routine screening analysis of Gent residues in food samples.     

A decision fusion method based on classification models for water quality monitoring

Environmental Science and Pollution Research - Monitoring of water quality is one of the world’s main intentions for countries. Classification techniques based on support vector machines...     

苯酚和间甲酚的竞争吸附研究

研究了水中苯酚和间甲酚在大孔树脂上的吸附,结果表明,ＩＡＳ及ＬＣＡ模型都能较好地反映这一吸附系统的竞争吸附规律。     

A fast and accurate method for specific detection and quantification of the bloom-forming microalgae Karlodinium veneficum in the marine environment

Environmental Science and Pollution Research - Karlodinium veneficum is a toxic benthic globally distributed dinoflagellate which has direct impacts on human health and the environment. Early and...     

A simple method to prepare magnetic modified beer yeast and its application for cationic dye adsorption

The purpose of this research is to use a simple method to prepare magnetic modified biomass with good adsorption performances for cationic ions. The magnetic modified biomass was prepared by two steps: (1) preparation of pyromellitic dianhydride (PMDA) modified biomass in N, N-dimethylacetamide solution and (2) preparation of magnetic PMDA modified biomass by a situ co-precipitation method under the assistance of ultrasound irradiation in ammonia water. The adsorption potential of the as-prepared magnetic modified biomass was analyzed by using cationic dyes: methylene blue and basic magenta as model dyes. Optical micrograph and x-ray diffraction analyses showed that Fe₃O₄ particles were precipitated on the modified biomass surface. The as-prepared biosorbent could be recycled easily by using an applied magnetic field. Titration analysis showed that the total concentration of the functional groups on the magnetic PMDA modified biomass was calculated to be 0.75 mmol g^?1 by using the first derivative method. The adsorption capacities (q _m) of the magnetic PMDA modified biomass for methylene blue and basic magenta were 609.0 and 520.9 mg g^?1, respectively, according to the Langmuir equation. Kinetics experiment showed that adsorption could be completed within 150 min for both dyes. The desorption experiment showed that the magnetic sorbent could be used repeatedly after regeneration. The as-prepared magnetic modified sorbent had a potential in the dyeing industry wastewater treatment.     

一种兼顾水质目标和成本投入的流域减排措施优化方法

综合考虑断面水质及治污成本,构建流域治污减排措施优化模型,分析单位污染物去除成本与断面水质改善间的关系,并以承德市武烈河流域治污减排优化为例,绘制治污减排措施优化曲线,提出优化方案。结果表明,在达到流域水质要求的情况下,优化后,治污减排项目由34项缩减到22项,平均单位COD去除成本由1.44万元/t下降到0.65万元/t,同时实现了流域水质达标和降低投入成本的要求。     

A modified in-vitro method to assess bioavailable arsenic in pesticide-applied soils

An incubation study was designed to modify the existing in vitro methods to increase the accuracy in estimation of bioavailable arsenic in pesticide-applied soils. In addition to simulating arsenic dissolution in gastric and intestinal solutions, absorbtion by the intestinal membrane was also mimicked using iron-oxide coated filter paper strips inserted in nylon bags. The in vitro experiments were sequentially performed in two phases, namely, the stomach phase and the absorbed-intestinal phase. Arsenic extraction in the in vitro absorbed intestinal phase increased, thereby making it more comparable to the potential in vivo arsenic pool. While animal studies are needed to verify the in vitro results, preliminary data indicate that this modified method may be able to improve site-specific bioavailability predictions in arsenic-contaminated soils.     

A new reliable method for dimethyl sulfoxide analysis in wastewater: dimethyl sulfoxide in Philadelphia's three water pollution control plants.

A simple but reliable procedure was developed to analyze dimethyl sulfoxide (DMSO) in wastewater. The isotope DMSO_d6 was used as the internal standard to ensure accuracy. The DMSO was reduced with stannous chloride and measured as dimethyl sulfide (DMS) with purge-and-trap gas chromatography/mass spectrometry. The method detection limit was at the sub-microgram-per-milliliter level; precision, as measured by standard deviation, was better than +/- 0.5%; and the recoveries were between 95 and 105% at the level of 2 microg/mL. The procedure could use standard analytical instrumentation used for volatile organic compound analysis. A field study was conducted to validate the method and quantify DMSO concentration range in the three water pollution control plants (WPCPs) in the city of Philadelphia, Pennsylvania. Results showed that, when a local chemical facility discharged, DMSO concentration could be as high as 12 mg/L in the influent to a WPCP. This would lead to the formation of a toxic "canned corn" DMS odor during the treatment processes.     

基于间接竞争抑制ELISA的2,2',4,4'-四溴联苯醚检测方法

针对现有的检测多溴联苯醚（PBDEs）方法中存在的样品处理繁复、样品检测耗时以及费用昂贵等不足,开展了针对此类物质中代表性化合物2,2',4,4'-四溴联苯醚（BDE-47）的间接竞争抑制酶联免疫吸附法（ELISA）的检测分析研究,并与气相色谱-质谱检测法、酶联免疫吸附分析试剂盒检测法进行对比分析。研究表明：间接竞争抑制ELISA法测定BDE-47的IC50值为0.11 mg·L-1,定量限为0.04 mg·L-1,检出范围为0.01~0.50 mg·L-1。该方法在满足复杂基质中微量目标物检测灵敏度及重现性要求下,具有检测更快捷、更经济实用的特点,宜适用于大批量此类环境样品的初期筛查。     

Bisphenol A concentrations in receiving waters near US manufacturing and processing facilities

Bisphenol A (BPA) (CAS 80-05-7) was analyzed in receiving waters upstream and downstream of US manufacturers (1996 and 1997) and processors (1997) during seasonal low flow periods. BPA was not detected (< 1 microgram/l) in any surface water sample in 1996 or at six of seven sites in 1997. Concentrations near the seventh site ranged from 2 to 8 micrograms/l; however, its receiving stream had no measurable flow and concentrations represent undiluted effluent. All surface water concentrations from this and other studies were less than the freshwater predicted no effect concentration (PNEC) of 64 micrograms/l, suggesting that BPA discharges from manufacturing and processing facilities to surface water do not pose an environmental concern.