Removal of Humic Substances from Aqueous Solution by Fungal Pellets

The removal of humic substances from aqueous solution at pH=4.8 and 6.0 and 25 °C by pure culture of Aspergillus niger has been examined. The removal of humic substances from aqueous phase was monitored by following the decrease in absorbance at 370 nm. COD and fungal growth were also measured. The initial concentration of humic and fulvic acids, pH and diameter of fungal pellets were found to be the most important parameters. The results suggested that decolorization of batch culture was a consequence of several processes. In the first 5 hours for pellets with diameter 1-3 mm and 10 hours for pellets with diameter 2-5 mm adsorption of humic substances was dominant. Afterwards, desorption of slightly bound humic substances and their degradation, as well as degradation of remaining humic substances in the solution, continued simultaneously through the period of 10 days.     

Application of Reflectance Spectroscopy for Analysis of Higher Plant Pigments

Nondestructive techniques developed by the authors for assessment of chlorophylls, carotenoids, and anthocyanins in higher plant leaves and fruits are presented. The spectral features of leaf reflectance in the visible and near infrared regions are briefly considered. For pigment analysis only reflectance values at several specific wavelengths are required. The chlorophyll (Chl) content over a wide range of its changes can be assessed during leaf ontogeny using reflectance near 700 nm and, in the absence of anthocyanins, at 550 nm. The approaches used for elimination of Chl interference in the analysis of carotenoids (reflectance at 520 nm) and anthocyanins (at 550 nm) are described. The suitability of reflectance spectroscopy for estimates of carotenoid/chlorophyll ratios during leaf senescence and fruit ripening is demonstrated. The algorithms developed for pigment analysis are presented, and the conditions of their applicability are considered. Further perspectives for the application of reflectance spectroscopy including remote sensing for estimation of plant pigment content and physiological states are discussed.     

基于近红外漫反射光谱快速测定淫羊藿蛋白质含量

采用近红外漫反射光谱技术对淫羊藿(Epimedium)的蛋白质含量进行快速且无损检测。近红外漫反射光谱经二阶导数处理、标准多元离散校正及主成分分析聚类处理后, 采用改进最小二乘法回归得到的定标模型预测效果最佳, 定标决定系数、交互验证标准差及交互验证相关系数分别为0.923、0.554和0.717。近红外光谱分析法的测定结果与用凯氏定氮法所得结果无显著差异, 两种方法测定值的相关性较高(R²=0.933 9)。重复性实验表明, 近红外光谱分析法的相对标准偏差为0.937%。该研究首次采用近红外光谱分析法测定了8种淫羊藿的蛋白质含量。该方法简便、精确, 在淫羊藿资源开发利用和药材质量控制方面具有参考意义。     

Characterizing Aeroallergens by Infrared Spectroscopy of Fungal Spores and Pollen

Background

Fungal spores and plant pollen cause respiratory diseases in susceptible individuals, such as asthma, allergic rhinitis and hypersensitivity pneumonitis. Aeroallergen monitoring networks are an important part of treatment strategies, but unfortunately traditional analysis is time consuming and expensive. We have explored the use of infrared spectroscopy of pollen and spores for an inexpensive and rapid characterization of aeroallergens.

Methodology

The study is based on measurement of spore and pollen samples by single reflectance attenuated total reflectance Fourier transform infrared spectroscopy (SR-ATR FTIR). The experimental set includes 71 spore (Basidiomycota) and 121 pollen (Pinales, Fagales and Poales) samples. Along with fresh basidiospores, the study has been conducted on the archived samples collected within the last 50 years.

Results

The spectroscopic-based methodology enables clear spectral differentiation between pollen and spores, as well as the separation of confamiliar and congeneric species. In addition, the analysis of the scattering signals inherent in the infrared spectra indicates that the FTIR methodology offers indirect estimation of morphology of pollen and spores. The analysis of fresh and archived spores shows that chemical composition of spores is well preserved even after decades of storage, including the characteristic taxonomy-related signals. Therefore, biochemical analysis of fungal spores by FTIR could provide economical, reliable and timely methodologies for improving fungal taxonomy, as well as for fungal identification and monitoring. This proof of principle study shows the potential for using FTIR as a rapid tool in aeroallergen studies. In addition, the presented method is ready to be immediately implemented in biological and ecological studies for direct measurement of pollen and spores from flowers and sporocarps.     

Real-Time UV-Visible Spectroscopy Analysis of Purple Membrane-Polyacrylamide Film Formation Taking into Account Fano Line Shapes and Scattering

We theoretically and experimentally analyze the formation of thick Purple Membrane (PM) polyacrylamide (PA) films by means of optical spectroscopy by considering the absorption of bacteriorhodopsin and scattering. We have applied semiclassical quantum mechanical techniques for the calculation of absorption spectra by taking into account the Fano effects on the ground state of bacteriorhodopsin. A model of the formation of PM-polyacrylamide films has been proposed based on the growth of polymeric chains around purple membrane. Experimentally, the temporal evolution of the polymerization process of acrylamide has been studied as function of the pH solution, obtaining a good correspondence to the proposed model. Thus, due to the formation of intermediate bacteriorhodopsin-doped nanogel, by controlling the polymerization process, an alternative methodology for the synthesis of bacteriorhodopsin-doped nanogels can be provided.     

基于NIRS法快速检测荞麦蛋白质与淀粉含量的研究

为建立近红外光谱技术测定荞麦蛋白质与淀粉含量的方法,本研究以217份荞麦样品为试验材料,采用最小二乘回归预测和交叉验证构建近红外预测模型。分析表明:前处理采用多元散射校正法(MSC),维数(Rank)分别为5和5,光谱区间6803.9~6094.2/cm所建立的荞麦蛋白质与淀粉含量模型的预测效果较好,其决定系数(R~2)分别为0.9481和0.9167,交叉验证均方根(RMSECV)分别为0.68和2.08,相对分析误差(RPD)分别为4.39和3.46,均大于3.0,外部验证相关系数均大于0.96。本试验所建立的蛋白质与淀粉含量近红外预测模型具有较高的准确度和稳健性,可用于荞麦品质的快速测定。     

近红外光谱仪快速检测油菜硫苷、芥酸及油份含量数学模型的建立

近红外反射光谱技术是一种高效、快速的现代分析技术,应用过程中需要建立相应的数学模型.收集油菜籽样品600余份,采用国际(家)标准方法对芥酸、硫苷、含油率进行测试,根据各模型的要求,选择不同的具有代表性的样品,建立数学模型.内部交叉证实法对模型验证结果显示:高芥酸、低芥酸、硫苷、含油率模型复相关系数分别为0.9677、0.9318、0.9820、0.9626,内部验证均方差(RMSECV)分别为4.41、2.24、4.62、0.543.外部样品检测法分别用已建模型和国际(家)标准方法对油菜籽样品芥酸、硫苷、含油率进行检测,两者检测结果基本一致.综合内部和外部验证结果表明:建立的模型能够满足油菜籽硫甘、芥酸及油份含量快速检测的要求.另对油菜自身含水率对数学模型的影响做了初步探索.     

近红外漫反射光谱法快速测定药用植物淫羊藿总黄酮含量

利用一种基于近红外漫反射光谱技术的淫羊藿总黄酮快速定量分析方法, 测定了淫羊藿属(Epimedium)7个种中总黄酮的含量。研究结果表明, 近红外光谱经二阶导数处理、主成分分析聚类及加权多元离散校正处理后, 采用改进最小二乘法回归得到的定标模型的预测效果最佳; 定标样品集预测方程的定标决定系数、交互验证标准差及交互验证相关系数分别为0.985 5、0.023 0和0.044 3; 检验样品集方程的定标决定系数、检验标准差及偏差分别为0.965 1、0.018和0.006, 达到了快速测定淫羊藿总黄酮含量的要求。该方法为淫羊藿总黄酮的快速定量分析提供了新的依据。     

生物组织的折射率对漫反射光分布的影响

采用杂合算法,讨论了折射率对不同吸收系数的生物组织漫反射光分布的影响。计算结果表明,在近源处生物组织的折射率越小,漫反射光模拟计算的相对误差越小;而在远离源处,其相对误差随折射率的增大而减小。同时也说明杂合模型不太适合模拟吸收生物组织的漫反射光分布。     

Rapid Biomass Quality Determination of Corn Stover Using Near-Infrared Reflectance Spectroscopy

Near-infrared reflectance spectroscopy (NIRS) has been used extensively in the forage industry for rapid measurement of forage constituents and could be useful for determining quality of biomass feedstocks at the point of delivery. In previous work, we developed an assay that partitions feedstock carbohydrates based on their availability to be converted to fermentable sugars, including non-structural carbohydrates (C _N), biochemically available carbohydrates (C _B) with an associated first-order availability rate constant (k _B), and unavailable carbohydrates (C _U ). Additional quality parameters measured included neutral detergent lignin (NDL), total available carbohydrates (C _A), and total carbohydrates (C _T). We evaluated the variability of biomass quality parameters in a set of corn stover samples and developed calibration equations for determining parameter values using NIRS. Fifty-two corn stover samples harvested in Iowa and Wisconsin in 2005 and 2006 were analyzed using a high-throughput assay for determining feedstock quality for biochemical conversion. Non-structural carbohydrates ranged from 84 to 155?g?kg^?1 dry matter (DM); C _B ranged from 354 to 557?g?kg^?1 DM; k _B ranged from 0.199 to 0.330?h^?1; C _A ranged from 463 to 699?g?kg^?1 DM, and NDL ranged from 32 to 74?g?kg^?1 DM. Significant differences (P?<?0.0001) among samples were observed for all parameters, except k _B. Near-infrared reflectance spectroscopy calibration equations were developed for C _N, C _B, C _A, C _U , C _T, and NDL. It was not possible to generate a meaningful calibration equation for k _B. There is significant variability within the corn stover population for several key quality-related carbohydrate and lignin constituents which can be predicted reliably using NIRS.     

Prediction of Soil Organic Carbon at the European Scale by Visible and Near InfraRed Reflectance Spectroscopy

Soil organic carbon is a key soil property related to soil fertility, aggregate stability and the exchange of CO₂ with the atmosphere. Existing soil maps and inventories can rarely be used to monitor the state and evolution in soil organic carbon content due to their poor spatial resolution, lack of consistency and high updating costs. Visible and Near Infrared diffuse reflectance spectroscopy is an alternative method to provide cheap and high-density soil data. However, there are still some uncertainties on its capacity to produce reliable predictions for areas characterized by large soil diversity. Using a large-scale EU soil survey of about 20,000 samples and covering 23 countries, we assessed the performance of reflectance spectroscopy for the prediction of soil organic carbon content. The best calibrations achieved a root mean square error ranging from 4 to 15 g C kg⁻¹ for mineral soils and a root mean square error of 50 g C kg⁻¹ for organic soil materials. Model errors are shown to be related to the levels of soil organic carbon and variations in other soil properties such as sand and clay content. Although errors are ∼5 times larger than the reproducibility error of the laboratory method, reflectance spectroscopy provides unbiased predictions of the soil organic carbon content. Such estimates could be used for assessing the mean soil organic carbon content of large geographical entities or countries. This study is a first step towards providing uniform continental-scale spectroscopic estimations of soil organic carbon, meeting an increasing demand for information on the state of the soil that can be used in biogeochemical models and the monitoring of soil degradation.     

Diffuse Reflectance Infrared Spectroscopic Identification of Dispersant/Particle Bonding Mechanisms in Functional Inks

In additive manufacturing, or 3D printing, material is deposited drop by drop, to create micron to macroscale layers. A typical inkjet ink is a colloidal dispersion containing approximately ten components including solvent, the nano to micron scale particles which will comprise the printed layer, polymeric dispersants to stabilize the particles, and polymers to tune layer strength, surface tension and viscosity. To rationally and efficiently formulate such an ink, it is crucial to know how the components interact. Specifically, which polymers bond to the particle surfaces and how are they attached? Answering this question requires an experimental procedure that discriminates between polymer adsorbed on the particles and free polymer. Further, the method must provide details about how the functional groups of the polymer interact with the particle. In this protocol, we show how to employ centrifugation to separate particles with adsorbed polymer from the rest of the ink, prepare the separated samples for spectroscopic measurement, and use Diffuse Reflectance Fourier Transform Infrared Spectroscopy (DRIFTS) for accurate determination of dispersant/particle bonding mechanisms. A significant advantage of this methodology is that it provides high level mechanistic detail using only simple, commonly available laboratory equipment. This makes crucial data available to almost any formulation laboratory. The method is most useful for inks composed of metal, ceramic, and metal oxide particles in the range of 100 nm or greater. Because of the density and particle size of these inks, they are readily separable with centrifugation. Further, the spectroscopic signatures of such particles are easy to distinguish from absorbed polymer. The primary limitation of this technique is that the spectroscopy is performed ex-situ on the separated and dried particles as opposed to the particles in dispersion. However, results from attenuated total reflectance spectra of the wet separated particles provide evidence for the validity of the DRIFTS measurement.     

基于光镊与拉曼光谱的红菇担孢子分析

拉曼光谱作为分子振动光谱,具有无损、无需试剂染色和快速方便的特点,在生物学领域的应用越来越广泛。拉曼镊子是光镊与拉曼光谱的结合,应用拉曼镊子技术收集单个红菇担孢子的拉曼光谱,分析单个担孢子的主要成分,不同的储存方法对担孢子成分的影响等。结果发现,单个担孢子的拉曼光谱反映了担孢子内含物的基本组成,红菇担孢子的孢内主要成分是脂类物质;不同种类的红菇担孢子的拉曼光谱基本相同,经过多年保存的孢子仍然保持其孢子内的主要成分不变。各随机选择10个担孢子的拉曼光谱进行主成分(PCA)判别分析,结果显示红菇属内不同种类的孢子无法区分。结果表明单孢子拉曼光谱可以应用于分析孢子内成分的变化,但应用于真菌分类的可能性有待进一步探索。     

近红外反射光谱分析技术在植物育种与种质资源研究中的应用

近年来近红外反射光谱分析技术(NIRS)在植物育种与种质资源研究中的应用已成为一个活跃的研究领域,本从NIRS分析植物的种类和品质类型、种质资源品质分析和评价、加速品质育种进程等方面作一综述,并对NIRS在作物育种中的应用作了探讨。     

Fungal Metabolism of Toluene: Monitoring of Fluorinated Analogs by 19F Nuclear Magnetic Resonance Spectroscopy

We used isomeric fluorotoluenes as model substrates to study the catabolism of toluene by five deuteromycete fungi and one ascomycete fungus capable of growth on toluene as the sole carbon and energy source, as well as by two fungi (Cunninghamella echinulata and Aspergillus niger) that cometabolize toluene. Whole cells were incubated with 2-, 3-, and 4-fluorotoluene, and metabolites were characterized by ¹⁹F nuclear magnetic resonance. Oxidation of fluorotoluene by C. echinulata was initiated either at the aromatic ring, resulting in fluorinated o-cresol, or at the methyl group to form fluorobenzoate. The initial conversion of the fluorotoluenes by toluene-grown fungi occurred only at the side chain and resulted in fluorinated benzoates. The latter compounds were the substrate for the ring hydroxylation and, depending on the fluorine position, were further metabolized up to catecholic intermediates. From the ¹⁹F nuclear magnetic resonance metabolic profiles, we propose that diverse fungi that grow on toluene assimilate toluene by an initial oxidation of the methyl group.     

144 Use of Fourier Transform Infrared (FT IR) Diffuse Reflectance Spectroscopy and Hamming Distances to Study the Phycocolloid Chemosystematics of the Red Algae (Rhodophta)

Coalescence in seaweeds is known to occur in the laboratory among young and older sporelings and in the field between neighboring conspecific clumps. However, because spores and germlings are difficult to study in the field, it is as yet unknown at which stage of population development coalescence is most important. Since many seaweeds disperse aggregated propagules, often with a sticky mucilagous envelope around the spores, aggregated recruitment and coalescence might be more important at early stages of population establishment than among fully grown, well established clumps. Using recruitment plates maintained during several experimental times in the field, we are evaluating the above idea with mid-intertidal populations of Mazzaella laminarioides. During high fertility seasons, close to 45% of the spores settling within or at close (<1 m) distances of the bed exhibited aggregated recruitment, forming groups of 2 to 150+spores. The probability of aggregated recruitments is a function of dispersal distance and spore density. The number of sporelings produced is a function of spore density and coalescence. Highest after-recruitment mortality (first 15 days) occurs among solitary recruits, followed by sporelings conformed by small number of spores (2–4). Approximately 50% of the spores recruited, isolated or in group, coalesce within these 15 days, gradually forming massive sporelings with increasingly larger basal areas. Thus, after recruitment, sporelings may disappear (die), survive or coalesce. These three alternatives are integrated in a new demographic model for coalescing seaweeds (supported by grant FONDECYT 1020855).     

Fungal Community Analysis by Large-Scale Sequencing of Environmental Samples

Fungi are an important and diverse component of soil communities, but these communities have proven difficult to study in conventional biotic surveys. We evaluated soil fungal diversity at two sites in a temperate forest using direct isolation of small-subunit and internal transcribed spacer (ITS) rRNA genes by PCR and high-throughput sequencing of cloned fragments. We identified 412 sequence types from 863 fungal ITS sequences, as well as 112 ITS sequences from other eukaryotic microorganisms. Equal proportions of Basidiomycota and Ascomycota sequences were present in both the ITS and small-subunit libraries, while members of other fungal phyla were recovered at much lower frequencies. Many sequences closely matched sequences from mycorrhizal, plant-pathogenic, and saprophytic fungi. Compositional differences were observed among samples from different soil depths, with mycorrhizal species predominating deeper in the soil profile and saprophytic species predominating in the litter layer. Richness was consistently lowest in the deepest soil horizon samples. Comparable levels of fungal richness have been observed following traditional specimen-based collecting and culturing surveys, but only after much more extensive sampling. The high rate at which new sequence types were recovered even after sampling 863 fungal ITS sequences and the dominance of fungi in our libraries relative to other eukaryotes suggest that the abundance and diversity of fungi in forest soils may be much higher than previously hypothesized.     

Ocular Injuries Caused by Airgun Pellets: An Analysis of 105 Cases

One hundred and five cases of airgun pellet injuries of the eye have been analysed. The average age was 14 and the male to female ratio 7·5:1. Forty-five patients had final visual acuities of 6/18 or less in the injured eye and 19 of these had the eye removed. The two main causes for poor vision were retinal damage and cataract. One patient became completely blind after sympathetic ophthalmitis arising 11 years after the injury. The pellet lodged in the orbit in 14 cases and three of these, together with one who had an intraethmoidal pellet, were investigated for abnormal lead levels in blood and urine. The results were normal.The circumstances of the injury were obtained in 12 cases and most involved careless handling of a gun. Instruction of children and parents in schools and on television, rather than stricter legislation, is suggested as a means of prevention.