Effect of fulvic and humic acids on nitrificationPart 1: In vitro production of nitrite and nitrate

Abstract

Mixed opinions exist on the effect of organic matter on nitrification in soils as well as the lack of data on the effect of fulvic (FA) and humic (HA) acids on this biochemical process. This in vitro investigation was conducted to study the effect of FA and HA on oxidation of NH⁺ ₄ and NO^‐ ₂ by soil nitrifiers and on the delay period (t') and maximum nitrification rate (K_max). Soil extracts containing an ammonium‐oxidizer population were incubated in vitro for 3 weeks at 25C in the presence of (NH₄)₂SO₄and 0 to 320 mg FA or HA/L at pH 7.0 or 8.0. A similar experiment was conducted with soil extracts containing a nitrite‐oxidizer population, but with KNO₂ as the N source. An additional experiment was conducted with the nitrite‐oxidizer extracts for the determination of t’ and K_max values. Nitrite production tended to increase linearly as a result of FA and HA treatments from 0 to 320 mg/L at pH 7.0 or 8.0. Fulvic acid appeared to be more effective than HA in increasing the oxidation process. Differences in pH had only a slight effect. On the other hand, nitrate production was decreased linearly by FA or HA treatments from 0 to 320 mg/L which provided some justification for reports of lower nitrate, but higher nitrite concentrations in soils high in organic matter content. Humic acid treatments increased the delay period (t¹), and at the same time decreased the maximum nitrification rate (K_max). The latter suggests that in the presence of HA more time is required to reach a maximum nitrification rate.     

Activity and stability of jack bean urease in the presence of peat humic acids obtained using different extractants

 The effects of two humic acid extractants, 0.1 M Na₄P₂O₇ plus 0.1 M NaOH (NaPP) and 0.1 M NaOH (NaOH), on the activity and stability of a humic-urease complex were studied. The two humic acids isolated (HA_NaPP and HA_NaOH) exhibited different elemental compositions, metal concentrations and structural modifications in the FT-IR spectra. Depending on the pH, HA_NaPP and HA_NaOH influenced both the urease activity and urease kinetic parameters (V _max and K _m) in the same way. They inhibited urease activity between pH 6 and 7 and reduced the V _max of the reaction at pH 6 and 7. The presence of humic acids improved the affinity of the enzyme for the substrate (K _m). The stability of the urease with time, and in the presence of pronase, was improved by HA_NaPP and HA_NaOH with respect to free enzyme. These results confirm the importance of the interaction of urease with humic acids as a fundamental gateway for extracellular urease stabilisation. Since no difference in the extent of urease inhibition and urease stabilisation was observed for the two humic acids, it may be concluded that neither urease activity nor stability are influenced by the humic acid extractant used. Received: 5 November 1999     

Influence of humic substances on photolysis of divalent mercury in aqueous solution

Mercury (II) solutions were irradiated by a simulated sunlight in the presence of humic acid (HA) or fulvic acid (FA). Results show that, under the experimental conditions and the FA and HA chosen, less than 20% of the Hg in solution was photolysed with a rate of (1.63±0.29)×10^?2 s^?1 (n=23) and the rest of (2.38±0.40)×10^?4 s^?1 (n=23) depending on the substitutes of humic substances to which Hg were bond. The sunlight photolysis lifetimes were estimated to be 4 and 250 sunlight hours respectively under summer conditions at Stockholm latitude.     

土壤中钙结合和铁铝结合腐殖质的组成及结构特征

Calcium-bound and iron- and aluminium-bound humus extracted from different soils collected from north to south of China were characterized by chemical and spectroscopic methods. Meaningful differences in the composition and structure between them were revealed by ¹³C NMR, visible spectroscopy and elemental analysis. Results showed that the contents of carbon, hydrogen and nitrogen were higher in iron- and aluminium-bound humus than in calcium-bound humus while oxygen content in calcium-bound humus was shown to be higher. The calcium-bound humus had higher C/N and O/C ratios than iron- and aluminium-bound humus. The calcium-bound humic acid (HA1) showed higher E₄/E₆ ratios than iron- and aluminum-bound humic acid (HA2) while iron- and aluminum-bound fulvic acid (FA2) showed higher E₄/E₆ ratios than calcium-bound fulvic acid (FA1). An inverse relationship between E₄/E₆ ratios and aromaticity as determined by ¹³C NMR spectra was observed for HA and FA from black soil. The ¹³C NMR spectroscopy revealed that HA2 was more aromatic than HA1. On the other hand, FA1 exhibited a higher aromaticity than FA2.     

Physiological Effects of Different Combinations of Humic and Fulvic Acid on Gerbera

Flower quality loss, especially short postharvest life, is a major problem in gerbera production. An experiment was conducted to determine how different combinations of humic substances (HS) affect gerbera. Humic acid (HA) and fulvic acid (FA) applied to nutrient solutions in six combinations including control (nutrient solution only), 80 mg L^?1 HA + 20 mg L^?1 FA, 60 mg L^?1 HA + 40 mg L^?1 FA, 40 mg L^?1 HA + 60 mg L^?1 FA, 100 mg L^?1 FA, and 50 mg L^?1 FA. The HS application enhanced root architecture, nutrient content, number of harvested flowers, and vase life. Fifty (50) mg L^?1 FA extended vase life by 8 days and increased flower number (72.9%). Results suggest that HA and FA (especially 50 mg L^?1 FA) can improve quality and quantity of gerbera through improving root architecture, leading to enhanced nutrient uptake and possibly affecting hormone-like activities. It seems that using low concentrations of FA may be part of a solution in improving gerbera flower quality.     

Dissolution of aluminum and iron phosphate by humic acids

Abstract

An investigation was conducted to study the effect of humic (HA) and fulvic acid (FA) on the dissolution of aluminum phosphate (AlPO₄) and iron phosphate (FePO₄), to analyze the dissolution products, and assess their availability to plants. The rate of dissolution was determined by shaking 10 mg of Al‐ or FePO₄ with 0 to 800 mg L^‐1 of HA or FA solutions at pH 7.0 for 0 to 192 hours. The phosphorus (P) concentration was measured in the extracts by spectrophotometry, whereas the nature of P‐humic acid complexes was determined by ³¹P NMR analysis. Availability of dissolution products was studied by growing corn plants in aerated hydroponic solutions receiving treatments of 50 mg Al‐ or FePO₄ and 0 to 800 mg L^‐1 of HA or FA at pH 5.0. The results indicated that the amount of P released by HA or FA increased with time. Humic acid was more effective than FA in dissolving the metal phosphates. The ³¹P NMR analysis showed that the dissolution products contained free orthophosphates and minor amounts of P‐humic acid complexes. This confirms the role of HA as a powerful chelator of Al and Fe, liberating in this way the orthophosphate anions. Corn plants grown in hydroponics, with AlPO₄ or FePO₄ as the source of P, exhibited better growth performance when HA or FA are present.     

Soil organic carbon storage and quality are impacted by corn cob biochar application on a tropical sandy loam

Purpose

Humic substances, which are integral components of total organic carbon (TOC), influence soil quality. The study aimed to investigate whether humic and non-humic fractions exhibit early, consistent, and measurable changes and affect TOC sensitivity and storage in a tropical sandy loam soils amended with corn cob biochar.

Materials and methods

There were four treatments with four replicates established in a randomized complete block design. Composite soil samples were taken from plots without biochar (CT), from plots incorporated with 15 t biochar ha^?1 (BC-15), and 30 t biochar ha^?1 without or with phosphate fertilizer (BC-30 and BC-30+P). The TOC, and humin, humic acid (HA), and fulvic acid (HA) fractions of soil organic carbon were determined for each treatment. The optical densities (400–700 nm) were measured on the soil-free extracts by spectrophotometry; the densities measured at 465 and 665 nm were used to calculate the E₄₆₅/E₆₆₅ ratios.

Results and discussion

The BC-30 and BC-30+P plots recorded the highest TOC, humin, humic acid (HA), and fulvic acid (FA) contents with respect to the lowest in the CT. The total exchangeable carbon stratification was significantly higher in all the biochar-treated plots relative to the CT. Spectral analysis showed higher values of E₄₆₅/E₆₆₅ (5.02 and 5.15) in the CT and BC-15-treated soils, respectively, compared with the BC-30 and BC-30+P-amended soils with E₄₆₅/E₆₆₅ ratios of 2.76 and 2.98, respectively.

Conclusions

Corn cob biochar applied to a tropical sandy loam:

? increased the concentrations of HA and FA and led to increased stratification of TOC, with a stronger effect on HA compared with FA;

? significantly lowered E₄₆₅/E₆₆₅ at the high biochar application rate of 30 t ha^?1, implying the dominance of high molecular weight humic acid-like substances, and increased degree of aromaticity of the TOC.

     

Kinetic behaviour of alkaline phosphatase desorbed from different clay minerals

The kinetic (K_m, V_max) of alkaline phosphatase (AP) desorbed from different Ca-homoionic clay minerals (montmorillonite, illite, and kaolinite) by extraction with Tris-Malate-Citrate buffer solution (pH 9.6) was studied in model experiments. After extraction (shaking for 15 min.) the K_m and V_max were measured in the extract, the remaining sediment and in the whole set-up. With kaolinite and illite, V_max of the desorbed AP was lower than that of the sediment. However, with montmorillonite, V_max of AP in the extract and whole system increased if compared to the control, but decreased in the sediment. The K_m of desorbed AP increased from 4.3 × 10^?3 (control) to 5.0 × 10^?3 M (illite), 5.4 × 10^?3M (kaolinite), and 5.5 × 10^?3M (montmorillonite). These values were lower than those obtained with the various sediments and whole experimental systems. An aberrant behaviour was recorded with the illite sorbed AP which showed an increase in affinity towards the substrate. Generally speaking, AP desorbed from clays may be reduced in its affinity towards the substrate p-nitrophenylphosphate by residual inhibitor and/or conformational change of the enzyme.     

Application of Near Infrared Reflectance Spectroscopy for the Assessment of Soil Quality in a Long‐Term Pasture

Abstract

The objective of the present study was to assess the ability of near infrared reflectance spectroscopy (NIRS) to analyze chemical soil properties and to evaluate the effects of different phosphorus (P) and potassium (K) fertilization rates on soil quality in different layers of a long‐term pasture. The NIRS calibrations were developed for humus, total Kjeldahl nitrogen (N_Kjeldahl), and several humic substances (HA1, “mobile” humic acids fraction; ΣHA, sum of humic acids; FA1, “mobile” fulvic acids; ΣFA, sum of fulvic acids, etc.) using soil samples of rather heterogeneous origin, collected during 1999–2003. Different spectral preprocessing and the modified partial least squares (MPLS) regression method were explored to enhance the relation between the spectra and measured soil properties. The equations were employed for the quality prediction of a sod gleyic light loam (Cambisol) in five PK fertilization treatments. The soil was sampled in 2000 and 2003 in three field replicates at depths of 0–10, 10–20, 20–30, and 30–50 cm, n=60 samples yr^?1. The best coefficients of correlation, R², between the reference and NIRS‐predicted data were as follows: for N_Kjeldahl, 0.965; humus, 0.938; HA1, 0.903; HA2, 0.905; HA3, 0.924; ΣHA, 0.904; and FA1, 0.911; and ΣFA, 0.885. Our findings suggest that it is feasible to use NIRS for the assessment of the effects of the inorganic PK fertilizer on the soil quality in different depths of a long‐term pasture.     

Peanut Straw Decomposition Products Promoted by Chemical Additives and Their Effect on Enzymatic Activity and Microbial Functional Diversity in Red Soil

ABSTRACT

The objectives of the present study were to determine the promotional effect of chemical additives on quality of peanut straw decomposition products and to evaluate the influence of the resulting products on soil biological properties. Straw was mixed with or without chemical additives, such as iron(II) sulfate (FeSO₄), alkali slag, or FeSO₄ combined with alkali slag, and decomposed for 50 days. The decomposition products were used as organic fertilizer and added to red soil for an incubation experiment. The chemical additives increased total organic carbon (C), total nitrogen (N), and available N content but decreased the C:N ratios in decomposition products compared to controls. Adding FeSO₄ gave the highest humic acid content (HA, 30.34 g kg^?1) and ratio of humic to fulvic acid (HA/FA, 0.53) and the lowest ratio of HA absorption value at 465 nm to that at 665 nm (E₄/E₆, 6.05), suggesting high humification of decomposition products. Application of the resulting products to soil increased soil urease and invertase activities. BIOLOG analysis showed that microbial C utilization ability, Shannon–Weaver diversity, and McIntosh evenness indexes were improved by the organic fertilizer promoted by chemical additives. Principal component analysis indicated that microbial community structures were also influenced by different amendments in decomposition products. Our study provides a reference point for acquiring high quality straw compost and improving soil biological functions by organic fertilizer.     

Inhibitory action of dissolved humic substances on the growth of soil bacteria degrading DDT

Abstract

Dissolved humic acid (HA) and fulvic acid (FA) prepared from a Dando brown forest soil (Typic Dystrochrept) inhibited the growth of soil bacteria degrading DDT (1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane) in the culture. The population of DDT-degrading Gram-variable rod Bll6 decreased by the application of both HA and FA, suggesting the presence of bactericidal effect. Such inhibitory effect was stronger for HA and resulted in a lower degrading activity of DDT in the culture of Bll6. No inhibitory effect was observed on the growth of DDT-degrading Bacillus sp. B75. The electron spin resonance spectra showed the presence of organic free radicals in both HA and FA. The relative concentration of the radicals was higher in HA. Storage of HA solution for 3 months at 4°C decreased the concentration of the radicals as well as the inhibitory action. The addition of catalase decreased the inhibitory effect of humic acid. It is suggested that a hydroxy radical, which is derived from free radicals of humic substances, is involved in the inhibition of bacterial growth and degradation of DDT.     

The significance of solid‐state 13C NMR spectroscopy of whole soil in the characterization of humic matter

Abstract

Differences in characteristics of humic matter were investigated by solid‐state CP/MAS ¹³C NMR spectroscopy of whole (nontreated) materials and their extracted humic fractions. Samples used in the analysis were lignite, a commercial humate AG, and the Bh horizons of a Mascotte and a Lawnwood soil. Humic fractions were extracted by the 0.1 M NaOH or Na₄P₂O₇ (pH 9.8) method. The humic (HA) and fulvic acid (FA) obtained were weighed and analysed for total acidity, carboxyl and phenolic‐OH group contents. Whole lignite, humate AG and soil samples, and the HA and FA fractions were analyzed by solid state CP MAS ¹³C NMR and infrared spectroscopy. Carbon, H, and N contents were determined by chemical analysis. NMR spectra of the combined HA+FA extracts resembled the spectra of the whole materials. No additional signals were detected, indicating that alien compounds were not produced during the extraction. The best spectra were obtained with HA samples produced by the NaOH method. These spectra closely resembled those of the untreated materials. Spectral and chemical differences noticed between the HA (or FA) fractions were attributed more to differences in origin than to the extraction procedure. Aliphatic, aromatic and carboxyl groups were the major components of HA from lignite and humate AG. In contrast, HA from the two Haplaquods were characterized by four major components: the aliphatic, polysaccharide, aromatic, and carboxyl groups. Regardless of origin, all the HA fractions contained similar functional groups, as indicated by their close similarities in infrared spectra.     

Anticlastogenic, antitoxic and sorption effects of humic substances on the mutagen maleic hydrazide tested in leguminous plants

The potential anticlastogenic and antitoxic effects of a soil humic acid (HA), a peat HA and a peat fulvic acid (FA) on the mutagen maleic hydrazide (MH) have been investigated in two legume species, Vicia faba and Pisum sativum. Both HAs and FA were tested at two different concentrations, 20 and 200 mg l^?1, either alone or after 24‐hour interaction with 10 mg l^?1 of MH before addition to the legume seeds. Anticlastogenicity, i.e. an antimutagenic action defined as the capacity for minimizing chromosome breakages, was evaluated by counting both micronuclei (MN) and aberrant anatelophases (AAT) in root‐tip cells. Length and dry weight of the seedling primary root were measured to test the antitoxic activity of HA and FA on MH. The possible occurrence and extent of adsorption or desorption of MH onto or from HA were also investigated. The two species responded differently to the anticlastogenic tests, with V. faba showing a greater number of MN and AAT anomalies than P. sativum. Peat HA and FA exhibited anticlastogenic and antitoxic activities of similar intensity and greater than those of soil HA. The adsorption capacity of both HAs for MH was small, thus suggesting that adsorption is not a major mechanism responsible for the reduction of clastogenicity and antitoxicity of MH by HA.     

Effect of soil depth on acid properties of humic substances extracted from an ombrotrophic peat bog in northwest Spain

The most southerly ombrotrophic peat bogs in Europe are in Galicia (northwest Spain). The humic matter in them originates from chemical processes in anaerobic conditions. We studied the acid properties of fulvic acids and humic acids isolated from two peat horizons of an ombrotrophic peat bog by potentiometric titration. Solutions containing 25, 50 and 100 mg l⁻¹ of each humic substance were titrated at ionic strengths 0.005 m , 0.01 m and 0.1 m (with KNO₃ as the inert electrolyte). Charge curves were analysed with a Donnan model to determine the intrinsic proton binding parameters. The concentration of the humic substance affected the charge curves more significantly at pH exceeding 6, and tended to disappear at greater concentrations. The proton binding conditional constants decreased with increasing ionic strength, this effect being more significant in the carboxylic groups with less affinity for protons. The proton binding constant of the carboxyl groups in a fulvic acid was one order of magnitude less than the value for the corresponding humic acid, whereas for the phenolic groups the values for both fractions were similar. The total content of acid groups was approximately 2 mol kg⁻¹ greater in the fulvic fraction than in the humic fraction. Both humic fractions from the lower horizon contained more acid groups than those from the upper horizon, mainly because the content of carboxyl groups increases with soil depth. Therefore, the humic substances in the lower horizon of the peat will be more negatively charged, which will affect their solubility and the binding of metal ions.     

Retention and release isotherm of arsenic in arsenic–humic/fulvic equilibrium study

Organic fractions from farm yard manure (FYM), vermicompost, municipal sludge, mustard cake, and surface soil of West Bengal, which was arsenic (As)-contaminated, were extracted and fractionated into fulvic and humic acid (FA and HA, respectively) fractions following standard procedures. These HA and FA samples were characterized by pH-potentiometric titrations, viscometric measurements and visible spectrophotometry. The stability constant (logK) of the complexes formed by these natural with As in aqueous phase was evaluated by the ion-exchange method. The logK values suggest that the organo-As complexes were quite stable. The release isotherm of As from the HA/FA complexes extracted from vermicompost and FYM was assayed in the presence of molybdate, nitrate, phosphate, sulfate and borate. The greatest tendency to displace As from the complexes was shown by sulfate, molybdite, and nitrate.     

Composition of humic acids extracted under air and nitrogen atmosphere

Abstract

Differences in nature and composition of humic matter, attributed to NaOH extraction under air or a N₂ gas atmosphere, were investigated. Samples from the Ap horizons of a Cecil and Onaway soil, and lignite were shaken with 0.1 M NaOH in the presence of air or under N₂ gas. The humic (HA) and fulvic acid (FA) separated were weighed, and analyzed for C, H, O, N, and S contents. Total acidity, carboxyl and phenolic‐OH group contents were determined by chemical analysis. Characterization was performed by ¹³C nuclear magnetic resonance (NMR) and infrared (IR) spectroscopy. The results indicated that differences in amounts of HA and FA, attributed to NaOH extraction under air and N₂ gas, were small and no definite trend was noted for more HA and FA extracted under air. No differences in C, H, O, N, and S contents of HA and FA were also noticed as a result of the influence of air or N₂ gas during extraction. However, HA and FA exhibited larger values for total acidities when extracted under a N₂ gas atmosphere than under air. The lower amounts of carboxyl and phenolic‐OH group contents in HA and FA as a result of NaOH extraction under air, however, do not support the idea of increased oxidation by NaOH. No differences were noticed in NMR and IR spectra of HA and FA attributable to the two extraction methods. The differences in NMR and IR features obtained were more the result of differences in origin than in methods of extraction.     

Soil Organic Matter Quality and Zinc and Lead Sorption as Affected by a Sewage Sludge Or a Sewage Sludge Compost Application

Sewage sludge (SS) or sewage sludge compost (SSC) were applied to soil under controlled conditions, at rates of 0 or 200 Mg ha^?1, to investigate changes in dissolved organic matter (DOM), humic acids (HA), and Pb and Zn sorption in the soil. Infrared spectroscopy, visible spectrophotometry, and sorption isotherms (mono-metal and competitive sorption systems) methods were used to assess the changer. The E₄/E₆ ratio (λ at 465 / λ at 665 nm) and the infrared spectra of DOM and HA showed aromatic behaviour in compost-soil (SSC-S); in contrast sewage sludge-soil (SS-S) showed an aliphatic behaviour. Application of either SS or SSC increased the Pb and Zn sorption capacity of soil. The Pb and Zn sorption increased in soil and soil mixtures with a competitive metal system. The metal affinity sequence for soil, SS-S, and SSC-S was compared with the predicted affinity sequences obtained from metal properties. Poor correspondence was observed between the metal affinity sequence and the metal affinity sequence predicted by ionic potential, indicating that metals bonding to soils were not predominantly electrostatic. An affinity sequence based on Pearson's theory agreed with the metal affinity sequences for soils. A statistical analysis showed that the bands assigned to esters (1080 cm^?1) of DOM, phenolic OH (1420 cm^?1), amide I (1650 cm^?1), carboxyl and carbonyl C=O stretches of different nature, C=O stretch of aromatic esters, aliphatic cetone, aldehyde (1720 cm^?1), ethers and esters (1230 cm^?1), aliphatic alcohols (1125 cm^?1), and lignin (1380 cm^?1) of HA were correlated with Zn constants of Langmuir adsorption isotherm (P < 0.05).     

Macromolecule‐P associations and inositol phosphates in some chilean volcanic soils of temperate regions

Abstract

Total organic P, humic and fulvic acid‐P associations and inositol phosphates in nine volcanic soils of southern Chile were determined. The concentration of organic P (Po) ranged from 654 to 1942 ppm accounting for 49% to 64% of total soil P. Phosphorus associated to humic (HA‐P) and fulvic acids (FA‐P) accounted for 51–68% and 32–49% of Po, respectively. Inositol penta‐ and hexaphosphates represented 42% to 67% of Po suggesting that significant amounts are associated with both humic and fulvic acids. Po content was significantly correlated to organic C, total soil P and HA‐P. HA‐P and FA‐P fractions obtained from the most representative soil were examined by dyalisis and gel filtration. While approximately 96% of HA‐P presented a molecular weight higher than 100,000 daltons, 53% of FA‐P had a molecular weight under 12,000 daltons. It is suggested that these more labile organic P forms would be more easily mineralized, thus increasing the available P pool.     

土壤腐殖质分组研究

采用Pallo法对两种耕地土壤腐殖质进行分组,初步研究了各组分的数量和结构特征。结果表明:焦磷酸钠提取的胡敏酸(HAp)是胡敏酸(HA)的主要组分;游离富里酸(FAf)是富里酸(FA)的主要组分;胡敏素(HM)的主要组分为非溶解性胡敏素(ISHM),而溶解性胡敏素(SHM)含量较低。通常随土层深度增加,HA、HM各组分的绝对数量和相对数量都下降;FA各组分的绝对数量也下降,而相对数量增加;HA/FA比值下降。一般来看,焦磷酸钠提取的富里酸(FAp)、氢氧化钠提取的富里酸(FAs)比相应的HAp、氢氧化钠提取的胡敏酸(HAs)的分子结构简单,铁结合胡敏素(HMi)比粘粒结合胡敏素(HMc)的分子结构简单;随土层深度增加,HAp、HAs的分子结构变简单,而HMi、HMc的分子结构变复杂。