Débora Marcondes Bastos Pereira Milori

HUMIFICATION DEGREE OF SOIL HUMIC ACIDS DETERMINED BY FLUORESCENCE SPECTROSCOPY

The humification process increases semiquinone-type free radical concentrations (SFRC) in humus. Their quantification by electron spin resonance spectroscopy (ESR) has been a good indicator of the degree of humification of soil organic matter. As an alternative to relatively complex and expensive ESR spectroscopy, fluorescence spectroscopy was used in this study to evaluate the humification degree of 18 humic acids (HA) extracted from four Brazilian soils under different land use, tillage, or cropping systems. Two fluorescence humification indexes of HA were calculated based on work done by Zsolnay et al. and Kalbitz et al. in 1999, and a third new fluorescence index was proposed. Our proposal is to use the blue wavelength (465 nm) as the HA fluorescence excitation source. As this wavelength is more resonant with the humificated groups present in soil HA samples, our hypothesis is that the resultant fluorescence will provide information about these structures and, therefore, on the degree of humification of the HA. The SFRC varied from 1.90 × 1017 to 14.75 × 1017 spins/g HA, characterizing a wide range of the degree of humification of soil HA. The lowest SFRC occurred in native forests (1.90 × 1017 to 7.50 × 1017 spins/g HA) and pasture soils (2.30 × 1017 to 4.64 × 1017 spins/g HA). In cultivated sites, soil HAs from no-tillage soil were less humified (2.41 × 1017 to 13.30 × 1017 spins/g HA) than those in conventionally tilled soil (5.68 to 14.75 × 1017 spins/g HA). The three fluorescence indexes show the same tendency as SFRC. Our fluorescence method was closely related to procedures found in the works of Zsolnay et al. and Kalbitz et al. (R∼0.9, P < 0.0001) and with SFRC (R∼0.85, P < 0.02), showing their potential as a simple and sensitive technique for evaluating the effect of land use and soil management systems on the humification degree of soil humic acids.

Characterization of humic acids from a Brazilian Oxisol under different tillage systems by EPR, 13C NMR, FTIR and fluorescence spectroscopy

The importance of soil organic matter functions is well known, but structural information, chemical composition and changes induced by anthropogenic factors such as tillage practices are still being researched. In the present paper were characterized Brazilian humic acids (HAs) from an Oxisol under different treatments: conventional tillage/maize-bare fallow (CT1); conventional tillage/maize rotation with soybean-bare fallow (CT2); no-till/maize-bare fallow (NT1); no-till/maize rotation with soybean-bare fallow (NT2); no-till/maize-cajanus (NT3) and no cultivated soil under natural vegetation (NC). Soil HA samples were analyzed by electron paramagnetic resonance (EPR), solid-state 13C nuclear magnetic resonance (13C NMR), Fourier transform infra-red (FTIR) and UV-Vis fluorescence spectroscopies and elemental analysis (CHNS). The FTIR spectra of the HAs were similar for all treatments. The level of semiquinone-type free radical determined from the EPR spectra was lower for treatments no-till/maize-cajanus (NT3) and noncultivated soil (1.74×1017 and 1.02×1017 spins g−1 HA, respectively), compared with 2.3×1017 spins g−1 HA for other soils under cultivation. The percentage of aromatic carbons determined by 13C NMR also decreases for noncultivated soil to 24%, being around 30% for samples of the other treatments. The solid-state 13C NMR and EPR spectroscopies showed small differences in chemical composition of the HA from soils where incorporation of vegetal residues was higher, showing that organic matter (OM) formed in this cases is less aromatic. The fluorescence intensities were in agreement with the percentage of aromatic carbons, determined by NMR (r=0.97 P<0.01) and with semiquinone content, determined by EPR (r=0.97 P<0.01). No important effect due to tillage system was observed in these areas after 5 years of cultivation. Probably, the studied Oxisol has a high clay content that offers protection to the clay–Fe–OM complex against strong structural alterations.

Evaluation of the effects of Candidatus Liberibacter asiaticus on inoculated citrus plants using laser-induced breakdown spectroscopy (LIBS) and chemometrics tools

This study investigated the organic and inorganic constituents of healthy leaves and Candidatus Liberibacter asiaticus (CLas)-inoculated leaves of citrus plants. The bacteria CLas are one of the causal agents of citrus greening (or Huanglongbing) and its effect on citrus leaves was investigated using laser-induced breakdown spectroscopy (LIBS) combined with chemometrics. The information obtained from the LIBS spectra profiles with chemometrics analysis was promising for the construction of predictive models to identify healthy and infected plants. The major, macro- and microconstituents were relevant for differentiation of the sample conditions. The models were then applied to different inoculation times (from 1 to 8 months). The models were effective in the classification of 82-97% of the diseased samples with a 95% significance level. The novelty of this method was in the fingerprinting of healthy and diseased plants based on their organic and inorganic contents.

Determinação da humificação da matéria orgânica de um latossolo e de suas frações organo-minerais.

Laser-induced fluorescence spectroscopy was used to determine the humification degree of the organic matter of a whole soil, that is, soil without any physical or chemical treatment. Particle size was determined in physically separated fractions. The humification of organic matter, the depth of the soil and the planting system can be correlated. On the surface of the soil, no tillage provided the smallest humification in comparison with the conventional tillage and reduced tillage. The fractions of the superficial layer of the soil (0-20 cm) indicated larger humification in the fraction from 20 to 53 µm and the smallest in the fraction of <2 µm.

Quantification of total carbon in soil using laser-induced breakdown spectroscopy: a method to correct interference lines

The C cycle in the Brazilian forests is very important, mainly for issues addressed to climate changes and soil management. Assessing and understanding C dynamics in Amazonian soils can help scientists to improve models and anticipate scenarios. New methods that allow soil C measurements in situ are a crucial approach for this kind of region, due to the costs for collecting and sending soil samples from the rainforest to the laboratory. Laser-induced breakdown spectroscopy (LIBS) is a multielemental atomic emission spectroscopy technique that employs a highly energetic laser pulse for plasma production and requires neither sample preparation nor the use of reagents. As LIBS takes less than 10 s per sample measurement, it is considered a promising technique for in situ soil analyses. One of the limitations of portable LIBS systems, however, is the common overlap of the emission lines that cannot be spectrally resolved. In this study a method was developed capable of separating the Al interference from the C emission line in LIBS measurements. Two typical forest Brazilian soils rich in Al were investigated: a spodosol (Amazon Forest) and an oxisol (Atlantic Forest). Fifty-three samples were collected and analyzed using a low-resolution LIBS apparatus to measure the intensities of C lines. In particular, two C lines were evaluated, at 193.03 and 247.86 nm. The line at 247.86 nm showed very strong interference with Fe and Si lines, which made quantitative analysis difficult. The line at 193.03 nm showed interference with atomic and ionic Al emission lines, but this problem could be solved by applying a correction method that was proposed and tested in this work. The line at 247.86 was used to assess the proposed model. The strong correlation (Pearsons coefficient R=0.91) found between the LIBS values and those obtained by a reference technique (dry combustion by an elemental analyzer) supported the validity of the proposed method.

Carbon stock and humification index of organic matter affected by sugarcane straw and soil management

The maintenance of sugarcane (Saccharum spp.) straw on a soil surface increases the soil carbon (C) stocks, but at lower rates than expected. This fact is probably associated with the soil management adopted during sugarcane replanting. This study aimed to assess the impact on soil C stocks and the humification index of soil organic matter (SOM) of adopting no-tillage (NT) and conventional tillage (CT) for sugarcane replanting. A greater C content and stock was observed in the NT area, but only in the 0-5 cm soil layer (p < 0.05). Greater soil C stock (0-60 cm) was found in soil under NT, when compared to CT and the baseline. While C stock of 116 Mg ha-1 was found in the baseline area, in areas under CT and NT systems the values ranged from 120 to 127 Mg ha-1. Carbon retention rates of 0.67 and 1.63 Mg C ha-1 year-1 were obtained in areas under CT and NT, respectively. Laser-Induced Fluorescence Spectroscopy showed that CT makes the soil surface (0-20 cm) more homogeneous than the NT system due to the effect of soil disturbance, and that the SOM humification index (HLIF) is larger in CT compared to NT conditions. In contrast, NT had a gradient of increasing HLIF, showing that the entry of labile organic material such as straw is also responsible for the accumulation of C in this system. The maintenance of straw on the soil surface and the adoption of NT during sugarcane planting are strategies that can increase soil C sequestration in the Brazilian sugarcane sector.

Phosphorus quantification in fertilizers using laser induced breakdown spectroscopy (LIBS): a methodology of analysis to correct physical matrix effects

The aim of this study was to develop a quantitative method to determine phosphorus in fertilizers of different matrix compositions using the laser induced breakdown spectroscopy (LIBS) technique. The LIBS spectra were acquired on 26 samples of organic and inorganic fertilizers by using a low cost, portable, gated CCD system in the atmospheric environment. Inductively coupled plasma optical emission spectroscopy (ICP-OES) was used as the reference technique. A method was developed to remove the outlier spectra and perform the baseline correction and peak normalization. By applying the proposed corrections, the linear correlation between LIBS and ICP increased from R = 0.76 to R = 0.95. An average error of 15% found in cross-validation of LIBS quantification appeared feasible for P quantification in fertilizers. Two reference samples with different matrix compositions were also analyzed, and the absolute error in the quantification was below 5%. Further, no significant fluctuation was found in P quantification when LIBS was performed over 150 days.

Infrared spectroscopy: A potential tool in huanglongbing and citrus variegated chlorosis diagnosis

Huanglongbing (HLB) and citrus variegated chlorosis (CVC) are serious threats to citrus production and have caused considerable economic losses worldwide, especially in Brazil, which is one of the biggest citrus producers in the world. Neither disease has a cure nor an efficient means of control. They are also generally confused with each other in the field since they share similar initial symptoms, e.g., yellowing blotchy leaves. The most efficient tool for detecting these diseases is by polymerase chain reaction (PCR). However, PCR is expensive, is not high throughput, and is subject to cross reaction and contamination. In this report, a diagnostic method is proposed for detecting HLB and CVC diseases in leaves of sweet orange trees using attenuated total reflectance Fourier transform infrared spectroscopy and the induced classifier via partial least-squares regression. Four different leaf types were considered: healthy, CVC-symptomatic, HLB-symptomatic, and HLB-asymptomatic. The results show a success rate of 93.8% in correctly identifying these different leaf types. In order to understand which compounds are responsible for the spectral differences between the leaf types, samples of carbohydrates starch, sucrose, and glucose, flavonoids hesperidin and naringin, and coumarin umbelliferone were also analyzed. The concentration of these compounds in leaves may vary due to biotic stresses.

Evaluation of laser induced breakdown spectroscopy for multielemental determination in soils under sewage sludge application.

Laser induced breakdown spectroscopy (LIBS) is an atomic emission spectroscopy technique for simple, direct and clean analysis, with great application potential in environmental sustainability studies. In a single LIBS spectrum it is possible to obtain qualitative information on the sample composition. However, quantitative analysis requires a reliable model for analytical calibration. Multilayer perceptron (MLP), an artificial neural network, is a multivariate technique that is capable of learning to recognize features from examples. Therefore MLP can be used as a calibration model for analytical determinations. Accordingly, the present study proposes to evaluate the traditional linear fit and MLP models for LIBS calibration, in order to attain a quantitative multielemental method for contaminant determination in soil under sewage sludge application. Two sets of samples, both composed of two kinds of soils were used for calibration and validation, respectively. The analyte concentrations in these samples, used as reference, were determined by a reference analytical method using inductively coupled plasma optical emission spectrometry (ICP OES). The LIBS-MLP was compared to a LIBS-linear fit method. The values determined by LIBS-MLP showed lower prediction errors, correlation above 98% with values determined by ICP OES, higher accuracy and precision, lower limits of detection and great application potential in the analysis of different kinds of soils.

Multiple response optimization of laser-induced breakdown spectroscopy parameters for multi-element analysis of soil samples.

Laser-induced breakdown spectroscopy (LIBS) is an emerging analytical technique to perform elemental analysis in natural samples independent of their physical state (solid, liquid, or gaseous). Due to its instrumental features, LIBS shows promising potential to perform analysis in situ and in environments at risk. Since the analytical performance of LIBS strongly depends on the choice of experimental conditions, each particular application needs a specific instrumental adjustment. The present study evaluated three LIBS instrumental parameters regarding their influences on signal-to-noise ratio (SNR) of seven elements in soil samples: laser pulse energy, delay time, and integration time gate. A multivariate technique was used due to the significant interaction among the evaluated parameters. Subsequently, to optimize LIBS parameters for each individual element response, a method for multiple response optimization was used. With only one simple screening design, it was possible to obtain a good combination among the studied parameters in order to simultaneously increase the SNR for all analytes. Moreover, the analysis of individual response for elements is helpful to understand their physical behavior in the plasma and also how they are embedded in the sample matrix.