Eleonora Peruzzi

Assessment of pollution impact on biological activity and structure of seabed bacterial communities in the Port of Livorno (Italy)

The main objective of this study was to assess the impact of pollution on seabed bacterial diversity, structure and activity in the Port of Livorno. Samples of seabed sediments taken from five selected sites within the port were subjected to chemical analyses, enzymatic activity detection, bacterial count and biomolecular analysis. Five different statistics were used to correlate the level of contamination with the detected biological indicators. The results showed that the port is mainly contaminated by variable levels of petroleum hydrocarbons and heavy metals, which affect the structure and activity of the bacterial population. Irrespective of pollution levels, the bacterial diversity did not diverge significantly among the assessed sites and samples, and no dominance was observed. The type of impact of hydrocarbons and heavy metals was controversial, thus enforcing the supposition that the structure of the bacterial community is mainly driven by the levels of nutrients. The combined use of chemical and biological essays resulted in an in-depth observation and analysis of the existing links between pollution macro-indicators and biological response of seabed bacterial communities.

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.

Biochemical activity and chemical-structural properties of soil organic matter after 17 years of amendments with olive-mill pomace co-compost.

This study evaluates soil fertility, biochemical activity and the soils ability to stabilize organic matter after application of composted olive-mill pomace. This organic amendment was applied in two different olive groves in southern Spain having different soil typologies (carbonated and silicic). Olive grove soils after 17 years of organic management with application of olive-mill pomace co-compost were of higher quality than those with conventional management where no co-compost had been applied. The main chemical parameters studied (total organic carbon, total nitrogen, available phosphorus, exchangeable bases, cation exchange capacity, total extractable carbon (TEC), and humic-to-fulvic acids ratio), significantly increased in soils treated with the organic amendment. In particular, the more resistant pool of organic matter (TEC) enhanced by about six and eight fold in carbonated and silicic soils, respectively. Moreover, the amended silicic soils showed the most significant increases in enzyme activities linked to C and P cycles (β-glucosidase twenty-five fold higher and phosphatase seven fold higher). Organic management in both soils induced higher organic matter mineralization, as shown by the higher pyrrole/phenol index (increasing 40% and 150% in carbonated and silicic soils, respectively), and lower furfural/pyrrole index (decreasing 27% and 71% in carbonated and silicic soils, respectively). As a result of mineralization, organic matter incorporated was also more stable as suggested by the trend of the aliphatic/aromatic index (decreasing 36% and 30% in carbonated and silicic soils, respectively). Therefore, management system and soil type are key factors in increasing long-term C stability or sequestration in soils. Thus application of olive-oil extraction by-products to soils could lead to important mid-to -long-term agro-environmental benefits, and be a valuable alternative use for one of the most widespread polluting wastes in the Mediterranean region.

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.

Phytoremediation of contaminated sediments: evaluation of agronomic properties and risk assessment

This study evaluates the possibility of reusing marine sediments in land management. The sediments, dredged from Livorno port (Italy), had previously been phytotreated, using a salt-tolerant plant cover and earthworms, with the aim of reducing the salt level and improving the texture and microbiology. In this study, sediments were investigated in order to: (1) test their capability to be used as a revitalised soil-like substrate (techno-soil), and (2) assess the human exposure risks associated with sediment management. Results obtained after 6 months of experiments performed with biological indicators composed of an association of graminae grass (Paspalum vaginatum), legumes (Trifolium alexandrinum) and earthworms (Eisenia foetida), showed that the substrate behaves like a natural soil capable of supporting biological life (total N=0.2%; total P=0.7%; EC=1.5 mS·cm −1; β−glucosidase=20 μ g PNP·g *h −1). It was also found that plants accumulate small amounts of heavy metals in shoot tissues (120 mg Zn·kg−1; 25 mg Cu·kg−1). In detail, risk analysis was performed considering: (1) sediment storage in a sealed disposal basin inside the Livorno port area, and (2) off-site phyto-remediation. The maximum hazard index was found for workers inside the port area, with values of 1.7 and 25 for dermal contact and vapour inhalation risks, respectively.

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).

Hydraulic and biochemical analyses on full-scale sludge consolidation reed beds in Tuscany (Italy)

The management of sewage sludge has recently become one of the most significant challenges in wastewater management. Reed bed systems appear to be an efficient and economical solution for sludge management in small wastewater treatment plants. Four years ago, one of the holding companies for water and wastewater in central Italy adopted this technology in 6 wastewater treatment plants. Hydraulic and biochemical analyses were performed on the most representative site to asses the behaviour of reed beds with regard to dewatering, mineralization and humification of disposed sludge. Moreover, daily water content analysis were performed in the interval between subsequent sludge loadings. Results indicated a decrease of sludge volume by about 93% on a yearly basis. Biochemical analysis highlighted that mineralization processes decrease over time due to a rapid decrease of microbial activity and labile substrates, such as DHase enzyme and water-soluble carbon and ammonium, respectively. Moreover, a significant interrelationship between the parameters linked with mineralization was found: after two years of operation, the process of mineralization of organic matter is still predominant in the humification of organic matter. Daily water content data were used to define a semi empirical equation describing the dynamics of the dewatering process. Overall, the use of sludge reed beds resulted feasible, ecologically sustainable and cost-effective.