Organic matter humification in olive oil mill wastewater by abiotic catalysis with manganese(IV) oxide

The chemical changes occurring in an olive oil mill wastewater (OMW) sample digested catalytically with MnO(2) for 30 and 60 days were evaluated comparatively with those occurring in the same OMW left standing for the same time in an open-air lagoon. Both treatments increased the pH and electrical conductivity and decreased the contents of dry matter, total organic C and total N, and C/N ratio of OMW. The humic acid (HA)-like fraction isolated from the fresh OMW was characterized by a marked aliphatic character, small O and acidic functional group contents, marked presence of proteinaceous materials, partially modified lignin moieties and polysaccharides-like structures, extended molecular heterogeneity, and small degrees of aromatic ring polycondensation, polymerization and humification. With increasing the time of either lagooning or catalytic digestion, a loss of aliphatic materials and an increase of extraction yield, oxygenation, acidic functional groups, carbohydrates and aromaticity occurred in the HA-like fractions. The more evident changes measured for the HA-like fractions from catalytically-digested OMW, with respect to those from lagooned OMW, indicated that MnO(2) was able to catalyze organic matter humification in OMW.     

Chemical characterization of humic substances extracted from organic-waste-amended soils

Humic substances were extracted from a soil treated, in a 4-year experiment, at different rates with a sludge from anaerobic treatment of combined civil and industrial wastes, and with agricultural manure. Elemental and chemical analyses, molecular weight (MW) distribution and infrared (IR) spectroscopy were performed on the purified humic acids (HA). Organic wastes significantly increased the HA content of the treated soils and improved CEC. The C/N, C/H and C/O ratios of HA extracted from the original wastes showed a higher degree of humification for sludge than for manure. This difference was also noticed for the C/N ratio of soil humic extracts, indicating a faster humification process for the sludges in soil. The content of oxygen-containing functional groups was lower than the ‘model’ HA reported in the literature, and even more so for HA from sludges, reflecting their anaerobic formation. MW distribution and E₄/E₆ ratios showed that the sludge material had a higher molecular complexity than manure, supporting the high degree of humification attributed to the former. HA extracted from sludge-treated soils revealed a molecular dimension increasing with the application doses of waste material. Infrared spectra showed that HA formed in soils after waste additions reflected the chemical composition of the original organic material, which was rich in aliphatic groups and peptides for sludge and in carbohydrates for manure. On the basis of this study, it is concluded that not only are organic waste additions able to build up the HA content in soils but the HA formed assume the chemical characteristics and the degree of humification of the original material.     

Description of characteristics of humic substances from different waste materials

Humic acids (HAs) extracted from different organic wastes have been characterised by chemical methods. The chemical properties of HAs showed differences depending on the source from which they were obtained. The C content in HAs from organic wastes (41.1-63.2%) fluctuated around the C value in soil HA with the exception of composted bark and tobacco dust. Compared with soil HA, the N contents of HAs from sewage sludge and brewery sludge were found much higher than the others. E4:E6 ratios for HAs in organic wastes were generally greater than that for soil HA, which indicated a low degree of condensation and humification. The carboxyl and phenolic-OH group contents ranged 0.51-2.23 and 11.1-20.7 meq g(-1), respectively. High values of carboxyl and phenolic-OH contents indicated that these materials were still within early stages of humification.     

Evolution of the fulvic acid fractions during co-composting of olive oil mill wastewater sludge and tree cuttings

Fulvic acids (FAs) were isolated by a conventional procedure from two mixtures of the sludge residue obtained from olive oil mill wastewater (OMW) evaporated in open-air pond and tree cuttings (TC) at different stages of the co-composting process. The FAs were analyzed for elemental (C, H, N, S, O) and acidic functional group (carboxylic and phenolic) composition, and by ultraviolet/visible, Fourier transform infrared and fluorescence spectroscopies. At the initial stage of composting, FAs from the OMW sludge-TC mixtures were characterized by a prevalent aliphatic character, large contents of C, S-containing groups, proteinaceous materials and polysaccharide components, extended molecular heterogeneity, small O and acidic functional group contents, and small degrees of aromatic ring polycondensation, polymerization and humification. As composting proceeded, C, H and S contents, C/N ratio, and aliphaticity decreased, whereas N, O, COOH and phenolic OH contents, C/H and O/C ratios, and aromaticity increased. These results suggested that, with increasing the composting time, the chemical and structural properties of the FA components of the two OMW sludge-TC mixtures approached the characteristics typical of native soil FAs. Thus, co-composting of OMW sludge mixed with TC may represent a suitable treatment for enhancing the quality of organic matter in these materials when used as soil amendments.     

Electrofocusing the compost organic matter obtained by coupling SEC-PAGE

Humic acids (HA)-like extracted from compost at the beginning (t(0)) and after 130 days of composting (t(130)) were fractionated by coupling size exclusion chromatography to polyacrylamide gel electrophoresis (SEC-PAGE). HA-like fractions with the same molecular size (MS) and electrophoretic mobility were pooled and further characterised by analytical polyacrylamide gel electrofocusing (EF) and compared with HA separated from a Typic Chernozem soil. During the composting process all fractions were subjected to quantitative and qualitative modifications: the high MS fraction was degraded, the mid MS fractions were qualitatively changed, the content of low MS fractions increased and changed qualitatively. The main changes in EF pattern of the non fractionated HA-like t(130) were associated to low MS fractions. Such data seem to be reliable for explanation what mechanisms and monitoring of the evolution of the compost organic matter for their agricultural uses.     

Effects of sewage sludge amendment on the properties of two Brazilian oxisols and their humic acids

The effect of sewage sludge (SS) amendment on the general properties of the top layers of a sandy and a clayey oxisols and on the nature of their humic acid (HA) fractions was evaluated by chemical and physico-chemical techniques. The amended soils, especially the sandy soil, benefited of SS amendment by increasing their pH to above neutrality and enhancing the contents of C, N, P, and Ca and cation exchange capacity. The SS-HA-like sample showed larger H and N contents and a greater aliphatic character and humification degree than the HAs isolated from non-amended soils. The composition and structure of amended soil HAs were affected by SS application as a function of soil type and layer. In particular, N-containing groups and aliphatic structures of SS-HA-like sample appears to be partially incorporated in the amended soil HAs, and these effects were more evident in the HAs from the sandy oxisol.     

Effects of the maturity of wood waste compost on the structural features of humic acids

An analytical scheme for the separation of humic substances (HSs) and non-humic substances (non-HSs) was established to estimate the humification index (HI), which was defined as the ratio of HS carbon content to non-HS carbon content. The alkaline compost-extract contained a mixture of HSs and non-HSs, while acidification of the compost-extract resulted in precipitation of humic acid (HA). The acidified supernatant contained fulvic acid (FA) and non-HSs. In the present study, DAX-8 resin was used to separate FA and non-HSs. HI values, which were estimated to evaluate the maturity of wood waste compost, increased with composting duration. To determine the effects of compost maturity on HA structural features, correlations between HI and indicators of the degree of HA humification (atomic ratios, acidic functional group contents, spectroscopic parameters and molecular weight) were investigated. HI values were significantly related to the indicators of the extent of HA humification during composting.     

Physico-chemical characteristics of humic substances of major soil associations of Bihar (India)

Humic acid (HA) and fulvic acid (FA) extracted from fourteen surface soil samples (0–20 cms) belonging to nine major Soil Association Groups scattered over different agro-climatic situations, were characterized by elemental and functional group analysis, E₄/E₆ ratios, coagulation behaviour and distribution of carbon in different soil humus fractions. The E₄/E₆ ratio of FA extracted from different soils was wider than that of HA. The coagulation behaviour of HA and FA fractions and also of cultivated and forest soils showed marked differences. The variations in the ratios of HAC:FAC (0.31 to 1.0) among different soils were indicative of the degree of humification under the influence of vegetation and agroclimatic conditions. The elemental composition of HA and FA, in general, indicated a higher carbon and nitrogen content and C/N ratio in the former than in the latter fraction. On the contrary, the oxygen content of FA was higher compared to that of HA. The carbon contents of HA extracted from the cultivated and forest soils of Hazaribagh were almost equal, as were also the carbon contents of HA from the cultivated and forest soils from Ranchi. Total acidity of FA of the soils selected in the present study was higher than that of HA. The functional groups such as carboxyl, phenolic hydroxyl and carbonyl were present in the two fractions in varying proportions.     

Chemical composition of fresh and composted urban waste

Chemical, spectroscopic (IR and NMR), and molecular characteristics of humus extracted from urban waste before and after compositing are reported. The main differences are in the contents of acidic groups and straight aliphatic chains which diminish in the composted material. In comparing humus characteristics of composted urban waste and soils, the only real difference was found in the elution curves on Sephadex G-100, where the ratio of 100,000 and 1,300 peaks was higher in the compost than in the soil curve. The finding suggests that this ratio is a parameter that could enable us, even if in a semiquantitative manner, to follow the humification process of the compost when its addition to the soil is not less than the autochthonous organic matter content.Research work supported by CNR, Italy, Special grant I.P.R.A.-Sub-project 1. Paper N. 1531.     

Recycling of organic wastes by employing Eisenia fetida

This paper reports the recycling of nutrients by vermicomposting of cow dung (CD), poultry droppings (PD) and food industry sludge (FIS) employing earthworms (Eisenia fetida). A total of six vermicomposting units were established and dynamics of chemical and biological parameters has been studied for 13 weeks. The waste mixture containing 50% CD+25% PD+25% FIS had better fertilizer value among studied waste combinations. At the end of experiment, vermicomposts showed decrease in pH and organic C, but increase in EC, total Kjeldhal N, total available P and total K contents. The C:N ratio of final vermicomposts also reduced to 10.7-12.7 from 22.8 to 56 in different waste combinations. The earthworms have good biomass gain and cocoon production in all vermicomposting units but CD alone and 50% CD+25% PD+25% FIS were better than other studied combinations.     

Humification processes of needle litters on forest floors in Japanese cedar (Cryptomeria japonica) and Hinoki cypress (Chamaecyparis obtusa) plantations in Japan

We quantitatively clarified the early humification processes on Japanese cedar and Hinoki cypress forest floors by using a litterbag experiment and the solid-state ¹³C CPMAS NMR technique. There was no significant effect on litter mass loss during early humification between both coniferous litters regardless of the shape of their needles. Carbon composition in both litters showed similar trends during early humification. A/O-A as a humification index was low, around 0.6, in both litters throughout the experiment period although 60% of litter mass was lost. Coniferous litter incubated for 3 years might not be well-humified and would be susceptible to physical fragmentation. Carbon mass loss rates in conifers were in the following order: O-alkyl > aliphatic > aromatic > carbonyl carbons, differing with hardwoods. Conifers had concomitantly higher and lower mass loss rates of aliphatic and aromatic carbons than hardwoods. Soil organic carbon (SOC) accumulated in topsoil for conifers had relatively high and low contents of aliphatic and aromatic carbons than that for hardwood. These compositional differences of SOC among forests could be caused by the high and low supply rates of aliphatic and aromatic carbons from litter to topsoil. Consequently, initial litter nature and humification processes can affect the compositional qualities of SOC accumulated in soil.     

Transformations of carbon and nitrogen in a Calciorthid soil amended with a range of organic residues

A calcareous soil was treated with an organic fertilizer mixture of grape debris and peat, or with pig manure, chicken manure, city refuse or with aerobic or anaerobic sewage sludges. Changes in different fractions of carbon and nitrogen were followed by analysis of the soil, immediately after these additions, after they had decomposed for six months in the soil, after a maize harvest, and after a subsequent barley harvest. The various forms of carbon had decreased after six months. The rate of decrease varied with the nature of the organic materials. In all samples, the fulvic acids fraction decreased most. The main transformations happened during the first six months whether there was a crop growing or not, but the crops had no influence on the breakdown of the organic materials. The ratio of humic acids/fulvic acids had increased after six months of humification and, in general terms, the final values of the ratio oxidizable carbon/extractable carbon were lower than the initial ones, indicating a higher degree of humification in the organic matter by the end of the experiment. The total nitrogen level remained practically constant after the first six months of humification even after the second crop. Part of the organic nitrogen of the soil samples amended with chicken manure, city refuse or the two sludges was transformed into mineral nitrogen during the humification process. The extractable nitrogen values increased because of the fertilizer applied during the pot trials.     

稻田土壤氧化态有机碳组分变化及其与甲烷排放的关联性

稻田土壤有机碳是甲烷排放的关键底物之一,不同研究者由于采取的有机碳研究方法不同而得出稻田甲烷排放与土壤有机碳关系的结论不一.为明确影响稻田甲烷排放的土壤有机碳组分,设计了稻田施用不同外源有机碳(稻草还田、鸡粪和猪粪)的田间试验,对稻田甲烷排放和土壤有机碳组分的动态变化及其关联性进行监测和分析.结果表明,猪粪处理的甲烷排放与化肥处理无显著差异,而鸡粪和稻草2个处理的甲烷排放分别比化肥增加1.67倍(P＜0.05),2.69倍(P＜0.05);甲烷排放量与土壤易氧化有机碳含量显示相同顺序:稻草＞鸡粪＞猪粪＞化肥;通径分析表明,土壤易氧化有机碳组分1(被33 mmol/L KMnO4氧化的有机碳)与甲烷排放直接相关,其他有机碳组分仅通过组分1间接作用于水稻生育后期甲烷排放,且排放量较低.由此推断,易氧化有机碳组分1是甲烷排放的主要底物,通过有效措施降低肥源中易氧化态有机碳组分1是减排甲烷的关键技术之一.     

东北次生杨桦林土壤碳氮动态特征

土壤分级组分是研究其碳氮动态的基础,次生杨桦林作为东北地区主要的天然林类型,目前相关数据的欠缺状态要求对此进行深入研究。为此,采集0—10cm、10—20cm、20—30cm长白山次生杨桦林土壤,通过土壤颗粒组分物理化学分级方法,将土壤分成5种组分:沙和稳定团聚体土壤组分(SA)、酸不溶土壤组分(AI)、易氧化土壤组分(EO)、颗粒态土壤组分(P)和可溶性土壤组分(S),进而分析了不同组分的质量分数、碳氮含量、碳氮分配比例及红外光谱5类官能团相对含量,旨在探讨次生杨桦林土壤固碳、氮供应机制。结果显示,接近90%的土壤质量集中在稳定组分AI(66.21%)和SA(22.11%)上,导致稳定组分中碳截获量最大(占土壤总碳量的2/3),而且其C/N比活跃组分(P和EO)大2—9倍;与碳不同,由于活跃组分中N含量比稳定组分大4—80倍,致使活跃组分P和EO氮的分配比例最大,分别占土壤总氮的33.1%和26.0%;除了占土壤质量很少的P和S外,组分间以及组分内的碳氮间多具有显著相关关系。这种土壤碳、氮在不同组分间贮存方式的差异使得土壤碳储存稳定性更高、而N肥力供应更快速。伴随不同组分碳氮储存的变化,不同组分间红外官能团存在显著差异,AI组分中绝大多数官能团相对含量均最低,而P和S组分中绝大多数官能团相对含量均较高,绝大多数官能团相对含量与碳含量、氮含量呈现显著的正相关关系,反映了官能团具有维持土壤碳氮的功能。同时,官能团与土壤C/N具有显著相关关系,反映出组分官能团相对含量的高低具有指示组分化学活性高低的作用。研究发现对于林分土壤的碳截获与氮供应的机制阐明具有重要的科学意义,这为深入了解东北次生杨桦林碳氮动态及对未来气候的响应提供基础数据。     

Effects of municipal waste compost and sewage sludge on proton binding behavior of humic acids from Portuguese sandy and clay loam soils

The effects of amendment with municipal solid waste compost (MSWC) and sewage sludge (SS) on acid-base properties of soil humic acids (HAs) were investigated. For this purpose, HAs were isolated from MSWC and SS and two different Portuguese soils, one sandy and the other clay loam, either unamended or amended with MSWC or SS at a rate of 60 t ha(-1), and analysed by potentiometric titrations at various ionic strengths (0.01, 0.05, 0.1 and 0.3M) over the pH range from 3.5 to 10.5. All titration data were fitted with the NICA-Donnan model and the variations of model parameters between the various HA samples were discussed. The HAs from MSWC and SS had lower acidic functional group contents and higher proton binding affinities than the control soil HAs. Amending soils with MSWC and SS determined a decrease of acidic functional group contents and an increase on proton binding affinities of soil HAs. These effects were more evident in SS-amended soil HAs than in MSWC-amended soil HAs, and in clay loam soil HA than in sandy soil HA.     

黄土高原典型区域土壤腐殖酸组分剖面分布特征

黄土高原作为典型的气候敏感带和生态环境脆弱区,诸多因素影响着这个区域的土壤有机碳及其组分的分布特征。本文以黄土高原典型区域土壤剖面 0—200 cm土样为对象,分析了土壤腐殖酸、胡敏酸（HA）和富里酸（FA）含量随地理位置及土层深度的分布特征,并进一步探讨了土壤腐殖酸、HA和FA与全氮含量及土壤颗粒组成的关系。结果表明,黄土高原主要类型土壤腐殖酸、HA和FA含量均较低,且存在明显地理位置和土层分异性：从南到北同层次土壤腐殖酸、HA和FA含量均显著降低,同一区域随土层深度增加各组分含量均表现为在 0—40 cm土层明显下降, 40—120 cm土层稍有下降,120 cm土层以下基本稳定;土壤腐殖酸占有机碳比例变化范围为26.6%—54.7%,相对较小,且在整个剖面变化幅度不大,从南向北土壤腐殖酸占有机碳比例有增加趋势;土垫旱耕人为土在 0—40 cm、 40—120 cm和 120—200 cm土层中HA占腐殖酸比例分别为39.8%、49.0%和53.5%,HA/FA分别为0.66、0.96和1.15,黄土正常新成土在以上土层中HA占腐殖酸比例分别为26.3%、33.9%和42.3%,HA/FA分别为0.36、0.51和0.73,干润砂质新成土在以上土层中HA占腐殖酸比例分别为13.4%、37.1%和45.2%,HA/FA分别为0.16、0.59和0.82,说明黄土高原南北主要类型土壤腐殖酸品质总体较差,均属富里酸型土壤,且从南到北腐殖酸品质逐渐下降;土壤腐殖酸、HA和FA均与全氮含量呈极显著线性相关（P<0.01）,土壤有机碳、腐殖酸及HA含量与粘粒及砂粒百分含量亦呈高度线性相关（P<0.01）。     

Characterization of natural organic matter (NOM) derived from sewage sludge compost. Part 1: chemical and spectroscopic properties

In this study changes in the properties of natural organic matter (NOM) were studied during composting of sewage sludge in a laboratory experiment using the pile method. Typical physicochemical parameters were measured during 53 days of composting including humic fractions. The effects of humification on the molecular properties of humic acids (HA) were investigated by 13C CP/MAS NMR spectroscopy. On the basis of chemical analyses, 53 days of composting sewage sludge with structural material can be divided into three phases: (i) domination of rapid decomposition of non-humic, easily biodegradable organic matter (two to three weeks), (ii) domination of organic matter humification and formation of polycondensed, humic-like substances (the next two weeks), (iii) stabilization of transformed organic material and weak microbial activity. Spectroscopic characterization (13C NMR) of compost humic acids reveals changes in their structures during maturation. The changes are highly correlated with the processes taking place in bulk compost.     

Feasibility of utilization of horse dung spiked filter cake in vermicomposters using exotic earthworm Eisenia foetida

This contribution reports the potential of vermicomposting technology in the management of horse dung (HD) spiked sugar mill filter cake (SMFC) using an epigeic earthworm Eisenia foetida under laboratory conditions. A total of six vermicomposters filled with different ratios of HD and SMFC were maintained for this study. The growth and fecundity of E. foetida was monitored for 12 weeks. Maximum growth was recorded in 90% HD+10% SMFC feed mixture containing vermicomposter. Earthworms' biomass gain and reproduction was favorably up to 50% HD+50% SMFC feed composition. Maximum cocoons were also recorded in 90% HD+10% SMFC feed mixtures, however increasing proportions of SMFC in different vermicomposters affected the growth and fecundity of worms. A significant decrease in C:N ratio and increase in total kjeldahl nitrogen, total available phosphorus and calcium contents was recorded. The heavy metals content was higher in the vermicompost obtained in all the reactors than initial feed substrates. Based on investigations it is concluded that vermicomposting could be an alternative technology for the management of filter cake if it is mixed in 1:1 ratio with horse dung.     

Humic acids of different origin as modifiers of cadmium-ion chemistry: A spectroscopic approach to structural properties and reactivity

The physical-chemical properties of humic acid fractions (HA) derived from urban sludge (CUS) and cattle manure (CCM) composts, and agricultural soil (FS) fertilised with sludge for ten years, were initially explored by elemental analysis, UV-Vis, FTIR and fluorescence. These properties were then compared with reference HA of terrestrial (SO) and aquatic (SR). To correlate the chemical properties and the reactivity of these HA, the binding of Cd(II) was investigated by fluorescence quenching techniques (FQ). Indeed, fluorescence spectroscopy has been proven to be a powerful tool in discriminating the origin, chemical features and degree of humification of naturally occurring organic matter. The HA compost exhibited higher N content, smaller molecular size and lower aromaticity than the reference HA. In addition, the CUS sample showed clear evidence of impurities, most likely of microbial origin, which was not evident in the FS sample (i.e. during its further evolution/humification in soil). The quenching effect of Cd(II) is adequately described by a modified Stern-Volmer equation, which is based on two population fluorophores, one not being accessible. The resulting Cd-HA logK (conditional association constants) decreased in the order SO > CUS > FS > CCM > SR, thus reflecting the relative binding affinity. A similar order was found for the corresponding Cd(II) capacity, which is based on total Cd content in Cd-humate precipitates. Lastly, fluorescence analysis of the soluble and insoluble fractions clearly revealed the fluorophores most involved in the binding process. In conclusion, our work provides evidence that compost is a reservoir of “humic-like” material capable of compensating for any organic carbon deficit in soil and lessening the effect of inorganic pollutants.     

Transformation of humic acids by two-domain laccase from Streptomyces anulatus

The properties of two-domain laccase of Streptomyces anulatus (SaSL) and its role in transformation of humic acids (HA) from chernozem, sod-podzolic soil and peat at alkaline pH values were studied. The SaSL was cloned, expressed in E. coli and obtained in an electrophoretically homogeneous state. SaSL is an oligomeric protein with a molecular weight of 235-300 kDa and six or eight subunits in the molecule. The molecular weight of the subunit is 37.7 kDa. Its catalytic properties are similar to those of the previously described two-domain laccase. The enzyme catalyzed oxidation of electron donors (K₄[Fe(CN)₆], ABTS) at acidic pH and phenolic substrates (2-methoxyphenol, 2,6-dimethixyphenol) at alkaline pH values. The efficiency of catalysis was higher in the case of electron donors than phenolic substrates. The enzyme showed a high thermal stability and was more stable at neutral and alkaline pH values. The enzyme effectively transformed humic acids at alkaline pH values. It was found that polymerization reactions took place during interaction of SaSL with HA, as well as with their high-molecular weight (>80 kDa) and low-molecular weight (<5 kDa) fractions. Our results suggest a possible involvement of the two-domaim laccases in humification processes in alkaline soils.