Maize yield response to a phosphorus-solubilizing microbial inoculant in field trials.

Findings from multi-year, multi-site field trial experiments measuring maize yield response to inoculation with the phosphorus-solubilizing fungus, Penicillium bilaiae Chalabuda are presented. The main objective was to evaluate representative data on crop response to the inoculant across a broad set of different soil, agronomic management and climate conditions. A statistical analysis of crop yield response and its variability was conducted to guide further implementation of a stratified trial and sampling plan. Field trials, analysed in the present study, were conducted across the major maize producing agricultural cropland of the United States (2005-11) comprising 92 small (with sampling replication) and 369 large (without replication) trials. The multi-plot design enabled both a determination of how sampling area affects the estimation of maize yield and yield variance and an estimation of the ability of inoculation with P. bilaiae to increase maize yield. Inoculation increased maize yield in 66 of the 92 small and 295 of the 369 large field trials (within the small plots, yield increased significantly at the 95% confidence level, by 0·17 ± 0·044 t/ha or 1·8%, while in the larger plots, yield increases were higher and less variable (i.e., 0·33 ± 0·026 t/ha or 3·5%). There was considerable inter-annual variability in maize yield response attributed to inoculation compared to the un-inoculated control, with yield increases varying from 0·7 ± 0·75 up to 3·7 ± 0·73%. No significant correlation between yield response and soil acidity (i.e., pH) was detected, and it appears that pH reduction (through organic acid or proton efflux) was unlikely to be the primary pathway for better phosphorus availability measured as increased yield. Seed treatment and granular or dribble band formulations of the inoculant were found to be equally effective. Inoculation was most effective at increasing maize yield in fields that had low or very low soil phosphorus status for both small and large plots. At higher levels of soil phosphorus, yield in the large plots increased more with inoculation than in the small plots, which could be explained by phosphorus fertilization histories for the different field locations, as well as transient (e.g., rainfall) and topographic effects.

Application of europium(III) chelate-dyed nanoparticle labels in a competitive atrazine fluoroimmunoassay on an ITO waveguide.

We have demonstrated the use of an optical indium tin oxide (ITO) (quartz) waveguide as a new platform for immunosensors with fluorescent europium(III) chelate nanoparticle labels (Seradyn) in a competitive atrazine immunoassay. ITO as a solid surface facilitated the successful use of particulate labels in a competitive assay format. The limit of detection in the new nanoparticle assay was similar to a conventional ELISA. The effect of particle size on bioconjugate binding kinetics was studied using three sizes of bioconjugated particle labels (107, 304, and 396nm) and a rabbit IgG/anti-IgG system in a 96-well plate. A decrease in particle size resulted in faster binding but did not increase the assay sensitivity. Flux calculations based on the particle diffusivity prove that faster binding of the small particles in this study was primarily due to diffusion kinetics and not necessarily to a higher density of antibodies on the particle surface. The results suggest that ITO could make a good platform for an optical immunosensor using fluorescent nanoparticle labels in a competitive assay format for small molecule detection. However, when used in combination with fluorescent particulate labels, a highly sensitive excitation/detection system needs to be developed to fully utilize the kinetic advantage from small particle size. Different regeneration methods tested in this study showed that repeated washings with 0.1 M glycine-HCl facilitated the reuse of the ITO waveguide.

Application of an enzyme-linked immunosorbent assay for the analysis of paraquat in human-exposure samples.

Paraquat is a toxic quaternary ammonium compound used as an herbicide around the world. Easy, fast, and inexpensive but sensitive methods are needed to study the effects of long-term, low-level exposure of paraquat on human health. An enzyme-linked immunosorbent assay (ELISA) was used for quantification of paraquat in urine and air-filter samples collected in a human-exposure study among farm workers in Costa Rica. A sample pretreatment consisted of removal of interfering substances using solid-phase extraction resin columns. The precision and accuracy of the method were tested using duplicate spiked urine samples. The correlation between results for blind samples obtained using ELISA and liquid chromatography-mass spectrometry was significant (R2 = 0.945 and 0.906 for spiked and field samples, respectively). With an LOQ of 2 ng mL(-1), this ELISA method was able to distinguish the exposed from the nonexposed farm workers. For the air-filter analysis, paraquat was extracted by 9 M H2SO4 at 60 degrees C for 12 hours, and the results obtained by ELISA showed good correlation (R2 = 0.918) with the spectrophotometric (256 nm) measurements. Paraquat in acid-stabilized urine samples was very stable, and no significant losses were detected during a 3-month storage at room temperature, at 4 degrees C, or at -20 degrees C.