Serena Doni

Effects of petroleum contamination on soil microbial numbers, metabolic activity and urease activity.

The influence of petroleum contamination on soil microbial activities was investigated in 13 soil samples from sites around an injection water well (Iw-1, 2, 3, 4) (total petroleum hydrocarbons (TPH): 7.5-78 mg kg(-1)), an oil production well (Op-1, 2, 3, 4, 5) (TPH: 149-1110 mg kg(-1)), and an oil spill accident well (Os-1, 2, 3, 4) (TPH: 4500-34600 mg kg(-1)). The growth rate constant (μ) of glucose stimulated organisms, determined by microcalorimetry, was higher in Iw soil samples than in Op and Os samples. Total cultivable bacteria and fungi and urease activity also decreased with increasing concentration of TPH. Total heat produced demonstrated that TPH at concentrations less than about 1 g kg(-1) soil stimulated anaerobic respiration. A positive correlation between TPH and soil organic matter (OM) and stimulation of fungi-bacteria-urease at low TPH doses suggested that TPH is bound to soil OM and slowly metabolized in Iw soils during OM consumption. These methods can be used to evaluate the potential of polluted soils to carry out self-bioremediation by metabolizing TPH.

Organic matter–microorganism–plant in soil bioremediation: a synergic approach

Bioremediation is a natural process, which relies on bacteria, fungi, and plants to degrade, break down, transform, and/or essentially remove contaminants, ensuring the conservation of the ecosystem biophysical properties. Since microorganisms are the former agents for the degradation of organic contaminants in soil, the application of organic matter (such as compost, sewage sludge, etc.), which increases microbial density and also provides nutrients and readily degradable organic matter (bioenhancement–bioaugmentation) can be considered useful to accelerate the contaminant degradation. Moreover, the organic matter addition, by means of the increase of cation exchange capacity, soil porosity and water-holding capacity, enhances the soil health and provides a medium satisfactory for microorganism activity. Plants have been also recently used in soil reclamation strategy both for their ability to uptake, transform, and store the contaminants, and to promote the degradation of organic contaminants by microbes at rhizosphere level. It is widely recognized that plant, through organic materials, nutrients and oxygen supply, produces a rich microenvironment capable of promoting microbial proliferation and activity.

Almond tree and organic fertilization for soil quality improvement in southern Italy.

The semi-arid Mediterranean region, characterized by long dry periods followed by heavy bursts of rainfall, is particularly prone to soil erosion. The main goal of this study is to evaluate the soil quality under different practices of bio-physical amelioration which involve the soil-plant system (almond trees) and microorganism-manure. This study, carried out in the South of Italy (Basilicata Region- Pantanello farm), considered two types of fertilization (mineral and organic) and three slope gradients (0, 2 and 6%), in order to evaluate the effects of management practices in resisting soil erosion. Chemical (organic carbon and nitrogen), physical (soil shrinkage and bulk density) and biochemical (dehydrogenase activity and hydrolytic enzyme activities) parameters were selected as markers to follow agro-ecological changes with time. The organic treatment affected soil microbiological and physico-chemical properties by increasing soil nutrient availability, microbial activity, and improving soil structure. The consistently higher values of the hydrolytic enzyme activities (β-glucosidase, phosphatase, urease and protease) often observed in the presence of plants and on the 0 and 2% slopes, suggested the stimulation of nutrient cycles by tree roots, which improve the conditions for soil microorganisms in carrying out their metabolic activity. In the 6% slope and, in particular, in the mineral fertilizer treatment, soil metabolism was lower as suggested by the dehydrogenase activity which was 50% lower than that found in the 0 and 2% slopes, this seemed to be related to a slowdown in the nutrient cycling and organic carbon metabolism. However, on this slope, in both mineral and organic treatments, a significant stimulation of hydrolytic enzyme activities and an improvement of soil structure (reduction of bulk density of about 10% and increase in total shrinkage from 20 to 60%) were observed with plants compared to the control soil. The combination of organic fertilization and almond trees resulted effective, also in the highest slope, in mitigating the degradation processes through the improvement of chemico-nutritional, biochemical and physical soil properties.

Exploring the functional soil-microbe interface and exoenzymes through soil metaexoproteomics

Functionally important proteins at the interface of cell and soil are of potentially low abundance when compared with commonly recovered intracellular proteins. A novel approach was developed and used to extract the metaexoproteome, the subset of proteins found outside the cell, in the context of a soil enriched with the nitrogen-containing recalcitrant polymer chitin. The majority of proteins recovered was of bacterial origin and localized to the outer membrane or extracellular milieu. A wide variety of transporter proteins were identified, particularly those associated with amino-acid and phosphate uptake. The metaexoproteome extract retained chitinolytic activity and we were successful in detecting Nocardiopsis-like chitinases that correlated with the glycoside hydrolase family 18 (GH18) chi gene data and metataxonomic analysis. Nocardiopsis-like chitinases appeared to be solely responsible for chitinolytic activity in soil. This is the first study to detect and sequence bacterial exoenzymes with proven activity in the soil enzyme pool.

In situ phytoremediation of a soil historically contaminated by metals, hydrocarbons and polychlorobiphenyls

In the past several years, industrial and agricultural activities have led to serious environmental pollution, resulting in a large number of contaminated sites. As a result, much recent research activity has focused on the application of bioremediation technologies as an environmentally friendly and economically feasible means for decontamination of polluted soil. In this study horse manure and Populus nigra (var. italica) (HM + P treatment) have been used, at real scale level, as an approach for bioremediation of a soil historically contaminated by metals (Pb, Cr, Cd, Zn, Cu and Ni) and organic contaminants, such as polychlorobiphenyls and petroleum hydrocarbon. After one year, the HM + P phytotreatment was effective in the reclamation of the polluted soil from both organic and inorganic contaminants. A reduction of about 80% in total petroleum hydrocarbon (TPH), and 60% in polychlorobiphenyls (PCBs) and total metals was observed in the HM + P treatment. In contrast, in the horse manure (HM) treatment, used as control, a reduction of only about 30% of TPH was obtained. In order to assess both effectiveness and evolution of the remediation system to a biologically active soil ecosystem, together with the pollution parameters, the parameters describing the evolution of the soil functionality (enzymatic activities and protein SDS-PAGE pattern) were investigated. A stimulation of the metabolic soil processes (increase in dehydrogenase activity) was observed in the HM + P compared to the HM treatment. Finally, preliminary protein SDS-PAGE results have permitted the identification of proteins that have been recovered in the HM + P soil with respect to the HM; this may become a basic tool for improving the biogeochemical status of soil during the decontamination through the identification of microbial populations that are active in soil decontamination.

Stabilisation and mineralisation of sludge in reed bed systems after 10-20 years of operation.

Sludge stabilisation and mineralisation during periods of operation between 10 and 21 years were investigated in three different systems receiving sludge from urban wastewater treatment plants situated in Denmark. Samples were taken along the entire profiles, in order to compare the effectiveness of the sludge stabilisation process. Particular attention was given to the stabilisation process occurring within the reed beds; in fact, parameters correlated to biochemical properties of organic sludge matter were determined. Statistical procedures were used to evaluate how the biochemical processes influence the quality of sludge organic matter. The level of total organic carbon and total nitrogen had a similar trend along the profile: their concentration decreased with increasing depth, reaching very low levels at the deepest layers. The same trend was also observed for the water-soluble carbon, N-NH(3), β-glucosidase and urease activities, and hydrolytic enzymes linked to C and N cycles: their values decreased dramatically with increasing depth, meaning that the level of mineralisation of the organic matter was higher in the deepest layers. The determination of extracellular enzymes bound to humic substances and humic carbon permitted evaluation of the stabilisation of organic sludge matter, and also allowed individuation of the ways in which the sludge was stabilised, in terms of mineralisation and humification of the organic matter.

Organic matter stabilization in reed bed systems: Danish and Italian examples.

In this study, results about sludge stabilization in reed bed systems (RBSs) after the entire period of operation in two different systems situated in Denmark (Helsinge 42,000 population equivalent (p.e.) - 10 years) and in Italy (La Fontina, 30,000 p.e. - 6 years) were presented. In order to evaluate the process of sludge stabilization, parameters that highlighted the biochemical and chemico-structural properties of organic sludge matter have been determined. The level of total and soluble nutrients, and enzyme activities, parameters related to overall microbial activity, showed that stabilization of the sludge similarly occurred in both RBSs, even though in different landscape ecosystems. The chemical-structural characterization of sludge organic matter highlighted how the processes of stabilization have occurred satisfactorily in both RBSs; in fact, significant levels of pyrolytic indices for mineralization and humification were reached. The successful stabilization of organic matter occurred in both RBSs and was confirmed by the absence of Escherichia coli, and also by the results of organic compounds (polycyclic aromatic hydrocarbons, linear alkyl benzene sulfonates, nonylphenol ethoxylates, di-2-ethylhexyl-phthalate) and heavy metals.

Use of earthworms (Eisenia fetida) to reduce phytotoxicity and promote humification of pre-composted olive oil mill wastewater

BACKGROUND Olive mill wastewaters (OMWW) contain a high recalcitrant organic load and an associated toxicity that make their treatment necessary before environmental application. The aim of this study was to evaluate the feasibility of promoting the valorization and reducing the phytotoxicity of OMWW through a pre-composting process together with straw-chip bulking materials followed by the application of earthworms (Eisenia fetida) in the presence of oat seedlings (Avena sativa L.) seedlings. RESULTS After 3 months, the pre-composted material showed properties similar to a partially digested compost with some significant amelioration of chemical-physical and biochemical properties. The application of earthworms permitted a significant decrease in chemical (total organic carbon, water-extractable organic carbon, total nitrogen) and biological parameters (dehydrogenase enzyme activity), and an increase in humic substances and available nitrogen forms. In the presence of plants a higher C/N ratio and a lower content of nitrates were observed. In addition, the reduction in phenolic compounds observed in treatments with earthworms caused a decrease in phytotoxicity by about 50% with respect to the pre-composted material, which results in an increase in germination index. CONCLUSION The utilization of earthworms, in particular in the presence of plants, may be an ecologically sound and economically feasible technology to obtain a non-toxic, high-value product useful for agricultural purposes.

Bioremediation of polluted soil through the combined application of plants, earthworms and organic matter

Two plant species (Paulownia tomentosa and Cytisus scoparius), earthworms (Eisenia fetida), and organic matter (horse manure) were used as an ecological approach to bioremediate a soil historically contaminated by heavy metals and hydrocarbons. The experiment was carried out for six months at a mesoscale level using pots containing 90 kg of polluted soil. Three different treatments were performed for each plant: (i) untreated planted soil as a control (C); (ii) planted soil + horse manure (20:1 w/w) (M); (iii) planted soil + horse manure + 15 earthworms (ME). Both the plant species were able to grow in the polluted soil and to improve the soils bio-chemical conditions, especially when organic matter and earthworms were applied. By comparing the two plant species, few significant differences were observed in the soil characteristics; Cytisus scoparius improved soil nutrient content more than Paulownia tomentosa, which instead stimulated more soil microbial metabolism. Regarding the pollutants, Paulownia tomentosa was more efficient in reducing the heavy metal (Pb, Cr, Cd, Zn, Cu, Ni) content, while earthworms were particularly able to stimulate the processes involved in the decontamination of organic pollutants (hydrocarbons). This ecological approach, validated at a mesoscale level, has recently been transferred to a real scale situation to carry out the bioremediation of polluted soil in San Giuliano Terme Municipality (Pisa, Italy).

Isoelectric focusing of β-glucosidase humic-bound activity in semi-arid Mediterranean soils under management practices

Changes in β-glucosidase enzyme–humic complexes and conventional parameters (pH, total organic C, total N, water-soluble C, and bulk density) were studied in an almond-cropped soil prone to erosion under a rehabilitation practice. The experimental plan included three soil slopes (0%, 2%, and 6%) and two type of fertilization (organic and mineral), with sampling of rhizosphere and inter-row soils. The enzyme humic complexes were extracted by pyrophosphate, purified by ultrafiltration of the organic extracts on molecular mass exclusion membranes (mol wt > 104) and fractionated by isoelectric focusing technique (IEF). The IEF on polyacrylamide rod gels with a restricted pH gradient ranging between 6.0 and 4.0 gave five humic bands on the basis of the little differences of their electric charges (pI). Under both organic and mineral fertilization, β-glucosidase activity bound to the fractionated humic substances, especially in the pH range 4.5–4.2 of the rhizosphere soil, was higher than that of the inter-row soil. This also occurred in 6% slope where the enzyme activity was lower than in soil with lower slopes. The higher number of the β-glucosidase active humic bands in rhizosphere than inter-row soil, particularly for the 0% slope, may be due to the presence of humic molecules capable of preserving the enzyme molecules in the active form, other than to the higher microbial activity synthesizing and releasing the tested enzymes.