Role of Agricultural Management in the Provision of Ecosystem Services in Warm Climate Vineyards: Functional Prediction of Genes Involved in Nutrient Cycling and Carbon Sequestration.

(1) Background: Maintaining soil fertility and crop productivity using natural microbial diversity could be a feasible approach for achieving sustainable development in agriculture. In this study, we compared soils from vineyards under organic and conventional management by predicting functional profiles through metagenomic analysis based on the 16S rRNA gene. (2) Methods: The structure, diversity and predictive functions of soil bacteria related to the biogeochemical cycle of the soil were analyzed, including oxidative and hydrolytic C-cycling enzymes, N-cycling enzymes and P-cycling enzymes. The inter-row spontaneous vegetation in the organic vineyards was also characterized. (3) Results: A clear effect of the farming system (organic vs. conventional) and cover management (herbicides plus tillage, mowing only and mowing plus tillage) on bacterial beta diversity and predicted functions was evidenced. While conventional viticulture increased the potential capacity of the soil to regulate the cycling of inorganic forms of N, organic viticulture in general enhanced those functions involving organic N, P and C substrates. Although the soil bacterial community responded differently to contrasting soil management strategies, nutrient cycling and carbon sequestration functions remained preserved, suggesting a high bacterial functional redundancy in the soil in any case. However, most of the predicted bacterial functions related to soil organic matter turnover were enhanced by organic management. (4) Conclusions: We posit the potential for organic viticulture to adequately address climate change adaptation in the context of sustainable agriculture.

Tomato Domestication Affects Potential Functional Molecular Pathways of Root-Associated Soil Bacteria.

While it has been well evidenced that plant domestication affects the structure of the root-associated microbiome, there is a poor understanding of how domestication-mediated differences between rhizosphere microorganisms functionally affect microbial ecosystem services. In this study, we explore how domestication influenced functional assembly patterns of bacterial communities in the root-associated soil of 27 tomato accessions through a transect of evolution, from plant ancestors to landraces to modern cultivars. Based on molecular analysis, functional profiles were predicted and co-occurrence networks were constructed based on the identification of co-presences of functional units in the tomato root-associated microbiome. The results revealed differences in eight metabolic pathway categories and highlighted the influence of the host genotype on the potential functions of soil bacterial communities. In general, wild tomatoes differed from modern cultivars and tomato landraces which showed similar values, although all ancestral functional characteristics have been conserved across time. We also found that certain functional groups tended to be more evolutionarily conserved in bacterial communities associated with tomato landraces than those of modern varieties. We hypothesize that the capacity of soil bacteria to provide ecosystem services is affected by agronomic practices linked to the domestication process, particularly those related to the preservation of soil organic matter.

Involvement of the metabolically active bacteria in the organic matter degradation during olive mill waste composting.

RNA-based high-throughput sequencing is a valuable tool in the discernment of the implication of metabolically active bacteria during composting. In this study, "alperujo" composting was used as microbial model for the elucidation of structure-function relationships with physicochemical transformation of the organic matter. DNA and RNA, subsequently retrotranscribed into cDNA, were isolated at the mesophilic, thermophilic and maturation phases. 16S rRNA gene was amplified by quantitative PCR (qPCR) and Illumina MiSeq platform to assess bacterial abundance and diversity, respectively. The results showed that the abundance of active bacteria assessed by qPCR was maximum at thermophilic phase, which confirm it as the most active stage of the process. Concerning diversity, Proteobacteria, Firmicutes, Bacteroidetes and Actinobacteria were the main phyla presented in composts. Concomitantly, three different behaviours were observed for bacterial dynamics: some genera decreased during the whole process meanwhile others proliferated only at thermophilic or maturation phase. Statistical correlation between physicochemical transformations of the organic matter and bacterial diversity revealed bacterial specialisation. This result indicated that specific groups of bacteria were only involved in the organic matter degradation during bio-oxidative phase or humification at maturation. Metabolic functions predictions confirmed that active bacteria were mainly involved in carbon (C) and nitrogen (N) cycles transformations, and pathogen reduction.

Resistance and Not Plant Fruit Traits Determine Root-Associated Bacterial Community Composition along a Domestication Gradient in Tomato.

Soil bacterial communities are involved in multiple ecosystem services, key in determining plant productivity. Crop domestication and intensive agricultural practices often disrupt species interactions with unknown consequences for rhizosphere microbiomes. This study evaluates whether variation in plant traits along a domestication gradient determines the composition of root-associated bacterial communities; and whether these changes are related to targeted plant traits (e.g., fruit traits) or are side effects of less-often-targeted traits (e.g., resistance) during crop breeding. For this purpose, 18 tomato varieties (wild and modern species) differing in fruit and resistance traits were grown in a field experiment, and their root-associated bacterial communities were characterised. Root-associated bacterial community composition was influenced by plant resistance traits and genotype relatedness. When only considering domesticated tomatoes, the effect of resistance on bacterial OTU composition increases, while the effect due to phylogenetic relatedness decreases. Furthermore, bacterial diversity positively correlated with plant resistance traits. These results suggest that resistance traits not selected during domestication are related to the capacity of tomato varieties to associate with different bacterial groups. Taken together, these results evidence the relationship between plant traits and bacterial communities, pointing out the potential of breeding to affect plant microbiomes.

Long-term no-tillage application increases soil organic carbon, nitrous oxide emissions and faba bean (Vicia faba L.) yields under rain-fed Mediterranean conditions.

The introduction of legumes into crop sequences and the reduction of tillage intensity are both proposed as agronomic practices to mitigate the soil degradation and negative impact of agriculture on the environment. However, the joint effects of these practices on nitrous oxide (N2O) and ammonia (NH3) emissions from soil remain unclear, particularly concerning semiarid Mediterranean areas. In the frame of a long-term field experiment (23 years), a 2-year study was performed on the faba bean (Vicia faba L.) to evaluate the effects of the long-term use of no tillage (NT) compared to conventional tillage (CT) on yield and N2O and NH3 emissions from a Vertisol in a semiarid Mediterranean environment. Changes induced by the tillage system in soil bulk density, water filled pore space (WFPS), organic carbon (TOC) and total nitrogen (TN), denitrifying enzyme activity (DEA), and bacterial gene (16S, amoA, and nosZ) abundance were measured as parameters potentially affecting N gas emissions. No tillage, compared with CT, significantly increased the faba bean grain yield by 23%. The tillage system had no significant effect on soil NH3 emissions. Total N2O emissions, averaged over two cropping seasons, were higher in NT than those in CT plots (2.58 vs 1.71 kg N2O-N ha-1, respectively; P < 0.01). In addition, DEA was higher in NT compared to that in CT (74.6 vs 18.6 µg N2O-N kg-1 h-1; P < 0.01). The higher N2O emissions in NT plots were ascribed to the increase of soil bulk density and WFPS, bacteria (16S abundance was 96% higher in NT than that in CT) and N cycle genes (amoA and nosZ abundances were respectively 154% and 84% higher in NT than that in CT). The total N2O emissions in faba bean were similar to those measured in other N-fertilized crops. In conclusion, a full evaluation of NT technique, besides the benefits on soil characteristics (e.g. TOC increase) and crop yield, must take into account some criticisms related to the increase of N2O emissions compared to CT.

Spider Communities and Biological Control in Native Habitats Surrounding Greenhouses.

The promotion of native vegetation as a habitat for natural enemies, which could increase their abundance and fitness, is especially useful in highly simplified settings such as Mediterranean greenhouse landscapes. Spiders as generalist predators may also be involved in intra-guild predation. However, the niche complementarity provided by spiders as a group means that increased spider diversity may facilitate complementary control actions. In this study, the interactions between spiders, the two major horticultural pests, Bemisia tabaci and Frankliniella occidentalis, and their naturally occurring predators and parasitoids were evaluated in a mix of 21 newly planted shrubs selected for habitat management in a highly disturbed horticultural system. The effects of all factors were evaluated using redundancy analysis (RDA) and the generalized additive model (GAM) to assess the statistical significance of abundance of spiders and pests. The GAM showed that the abundance of both pests had a significant effect on hunter spider's abundance, whereas the abundance of B. tabaci, but not F. occidentalis, affected web-weavers' abundance. Ordination analysis showed that spider abundance closely correlated with that of B. tabaci but not with that of F. occidentalis, suggesting that complementarity occurs, and thereby probability of biocontrol, with respect to the targeted pest B. tabaci, although the temporal patterns of the spiders differed from those of F. occidentalis. Conservation strategies involving the establishment of these native plants around greenhouses could be an effective way to reduce pest populations outdoors.

Long-term effects of contrasting tillage on soil organic carbon, nitrous oxide and ammonia emissions in a Mediterranean Vertisol under different crop sequences.

This 2-year study aimed to verify whether the continuous application of no tillage (NT) for over 20years, in comparison with conventional tillage (CT), affects nitrous oxide (N2O) and ammonia (NH3) emissions from a Vertisol and, if so, whether such an effect varies with crop sequence (continuous wheat, WW and wheat after faba bean, FW). To shed light on the mechanisms involved in determining N-gas emissions, soil bulk density, water filled pore space (WFPS), some carbon (C) and nitrogen (N) pools, denitrifying enzyme activity (DEA), and nitrous oxide reductase gene abundance (nosZ gene) were also assessed at 0-15 and 15-30cm soil depth. Tillage system had no significant effect on total NH3 emissions. On average, total N2O emissions were higher under NT (2.45kgN2O-Nha-1) than CT (1.72kgN2O-Nha-1), being the differences between the two tillage systems greater in FW than WW. The higher N2O emissions in NT treatments were ascribed to the increased bulk density, WFPS, and extractable organic C under NT compared to CT, all factors that generally promote the production of N2O. Moreover, compared to CT, NT enhanced the potential DEA (114 vs 16µgNkg-1h-1) and nosZ gene abundance (116 vs 69 copy number mg-1 dry soil) in the topsoil. Finally, NT compared to CT led to an average annual increase in C stock of 0.70MgCha-1year-1. Though NT can increase the amount os soil organic matter so storing CO2 into soil, some criticisms related to the increase of N2O emission arise, thereby suggesting the need for defining management strategies to mitigate such a negative effect.

Bottom-up effects on herbivore-induced plant defences: a case study based on compositional patterns of rhizosphere microbial communities.

Below-ground soil microorganisms can modulate above-ground plant-insect interactions. It still needs to be determined whether this is a direct effect of single species or an indirect effect of shifts in soil microbial community assemblages. Evaluation of the soil microbiome as a whole is critical for understanding multi-trophic interactions, including those mediated by volatiles involving plants, herbivorous insects, predators/parasitoids and microorganisms. We implemented a regulated system comprising Nerium oleander plants grown in soil initially containing a sterile/non sterile inoculum, herbivore Aphis nerii and predator Chrysoperla carnea. After aphid attack, plants emitted a characteristic blend of volatiles derived from two biosynthetic classes: fatty acid catabolites and aromatic-derived products. Three aliphatic compounds were mainly detected in plants grown in the inoculated microbial soil, a blend which was preferentially chosen by C. carnea adult females. The contrasting effect of the initial inocula was attributed to the different microbial consortia developed in each treatment. We argue that differences in the relative abundance of the active microbial communities in the rhizosphere correlate with those in the emission of selected volatile compounds by attacked plants. The mechanisms involved in how the functional soil microbiome modulates inducible indirect defence of plants are discussed.

Impact of agricultural management on bacterial laccase-encoding genes with possible implications for soil carbon storage in semi-arid Mediterranean olive farming.

BACKGROUND: In this work, we aimed to gain insights into the contribution of soil bacteria to carbon sequestration in Mediterranean habitats. In particular, we aimed to use bacterial laccase-encoding genes as molecular markers for soil organic C cycling. Using rainfed olive farming as an experimental model, we determined the stability and accumulation levels of humic substances and applied these data to bacterial laccase-encoding gene expression and diversity in soils under four different agricultural management systems (bare soils under tillage/no tillage and vegetation cover under chemical/mechanical management). MATERIALS AND METHODS: Humic C (> 10(4) Da) was subjected to isoelectric focusing. The GC-MS method was used to analyze aromatic hydrocarbons. Real-Time PCR quantification and denaturing gradient gel electrophoresis (DGGE) for functional bacterial laccase-like multicopper oxidase (LMCO)-encoding genes and transcripts were also carried out. RESULTS: Soils under spontaneous vegetation, eliminated in springtime using mechanical methods for more than 30 years, showed the highest humic acid levels as well as the largest bacterial population rich in laccase genes and transcripts. The structure of the bacterial community based on LMCO genes also pointed to phylogenetic differences between these soils due to the impact of different management systems. Soils where herbicides were used to eliminate spontaneous vegetation once a year and those where pre-emergence herbicides resulted in bare soils clustered together for DNA-based DGGE analysis, which indicated a certain amount of microbial selection due to the application of herbicides. When LMCO-encoding gene expression was studied, soils where cover vegetation was managed either with herbicides or with mechanical methods showed less than 10% similarity, suggesting that the type of weed management strategy used can impact weed community composition and consequently laccase substrates derived from vegetation decay. CONCLUSIONS: We suggest that the low humic acid content retrieved in the herbicide-treated soils was mainly related to the type (due to vegetal cover specialization) and smaller quantity (due to lower vegetal biomass levels) of phenolic substrates for laccase enzymes involved in humification processes. We also found that spontaneous vegetal cover managed using mechanical methods could be the best option for achieving C stabilization in rainfed Mediterranean agroecosystems.

Molecular tools to understand the bioremediation effect of plants and earthworms on contaminated marine sediments.

A meso-scale pilot plant was set up to test the efficiency of a bioremediation scheme applied to marine sediments contaminated by heavy metals and hydrocarbons. The experiment was implemented for three years in two stages using two remediation agents: plants (Paspalum vaginatum and Tamarix gallica) and earthworms (Eisenia fetida). DNA and RNA-based methodologies were applied to elucidate the dynamics of the bacterial population and were related to improving biological and chemical conditions of the sediments. Bioremediation strategies were successful in removing pollutants from the contaminated sediments and specialization within the bacterial community related to the type of contamination present was detected in the different stages of the process. The highest response of Gram-positive PAH-degraders to the contamination was detected at the beginning and after the first stage of the experiment, corresponding to the uppermost values of degradation.

Bacterial ß-glucosidase function and metabolic activity depend on soil management in semiarid rainfed agriculture.

Genomic and transcriptomic approaches were used to gain insights into the relationship between soil management and bacterial-mediated functions in an olive orchard agroecosystem. Four management practices were assessed in a 30-year trial in a semiarid Mediterranean region. Transcriptional activity of bacterial 16S rRNA genes increased in noncovered soils, indicating higher microbial maintenance requirements to thrive in less favorable environmental conditions. The 16S rRNA transcript:gene copy ratio confirmed this assumption and pointed toward a much higher constitutive expression from rRNA operons in noncovered soils and to even higher expression levels when spontaneous vegetation was removed chemically. As described for 16S rRNA, potential transcription did not reveal the real transcription of bacterial ß-glucosidase genes, and higher gene expression in noncovered soils plus herbicides was evidenced. Since no relationship between total or soluble organic carbon and bacterial ß-glucosidase transcription was found, the above hypothesis could indicate either that soluble organic carbon is not the main pool of enzyme-inducing substrates or that constitutive production of bacterial ß-glucosidase enzymes increases as soil conditions worsen.

Dynamic changes in bacterial community structure and in naphthalene dioxygenase expression in vermicompost-amended PAH-contaminated soils.

The aim of the present study was to explore the potential for using vermicompost from olive-mill waste as an organic amendment for enhanced bioremediation of polycyclic aromatic hydrocarbons (PAHs)-contaminated soils. The focus was to analyse the genetic potential and the naphthalene dioxygenase (NDO) expression of the bacterial communities involved in the degradation of naphthalene, as chemical model for the degradation of PAH. The structure of the metabolically active bacterial population was evidenced in the RNA-based denaturing gradient gel electrophoresis (DGGE) profiles. The relative expression of NDO was determined with real-time PCR in both the soil and the vermicompost cDNA. Naphthalene changed the structure of the metabolically active bacterial community in the vermicompost when this was artificially contaminated. When used as amendment, naphthalene-free vermicompost modified the bacterial population in the PAH-contaminated soil, evidenced in the DGGE gels after 1 month of incubation. In the amended soil, the vermicompost enhanced the NDO enzyme expression with a concomitant biodegradation of naphthalene. The effect of the vermicompost was to induce the expression of biodegradation indicator genes in the autochthonous bacterial community and/or incorporate new bacterial species capable of degrading PAH. The results indicated that vermicompost from olive-mill wastes could be considered a suitable technology to be used in PAH bioremediation.

Solvent tolerance acquired by Brevibacillus brevis during an olive-waste vermicomposting process.

In this work, a cultivable, Gram-positive, solvent-resistant bacterium was isolated from vermicomposted olive wastes (VOW). The highest 16S rRNA sequence similarity (99%) was found in Brevibacillus brevis. The genome of the isolate, selected for trichloroethylene (TCE)-tolerance, contained a nucleotide sequence encoding a conserved protein domain (ACR_tran) ascribable to the HAE1-RND family. Members of this family are hydrophobic/amphiphilic efflux pumps largely restricted to Gram-negative bacteria. No DNA sequences of HAE1 transporters were detected in the genome of a reference B. brevis strain isolated from natural soil. Since no cultivable solvent-tolerant bacterium was detected in the unvermicomposted olive waste, a transfer of solvent-resistance genes from Gram-negative bacteria during the vermicomposting process could explain the presence of HAE1 transporters in B. brevis isolated from the vermicompost. Under TCE stress conditions, the acquired nucleotide sequence could be translated into proteins, and the tolerance to solvents is conferred to the bacterium. The isolate was designated as strain BEA1 (EF079071).

Bioconversion of wastes from olive oil industries by vermicomposting process using the epigeic earthworm Eisenia andrei.

The present work evaluates the possible bioconversion of wet olive cake by low-cost biostabilization (vermicomposting process). Wet olive cake fresh (WOC), precomposted (WOCP), or mixed with biosolids (WOCB), were vermicomposted for 6 months to obtain organic amendments for agricultural and remediation purposes. The results showed initial differences depending on previous treatment. WOCP was initially more stable, presented a low C:N ratio, and showed more dehydrogenase and urease activity. By contrast, there was no dehydrogenase activity initially in WOC and WOCB, due to the presence of some different types of polyphenols. Finally, the end product showed relatively higher amounts of total nitrogen and humic acid and met the standard of quality for composts and vermicomposts for use both in conventional and organic agriculture and soil-restoration programs.

Restoring biochemical activity and bacterial diversity in a trichloroethylene-contaminated soil: the reclamation effect of vermicomposted olive wastes.

BACKGROUND, AIM, AND SCOPE: In this work, the potential for using olive-mill solid waste as an organic amendment for biochemical and biological restoration of a trichloroethylene-contaminated soil, which has previously been stabilized through vermicomposting processes, has been explored. MATERIALS AND METHODS: Trichloroethylene-contaminated water was pumped into soil columns with a layer of vermicompost at 10-cm depth (biobarrier system). The impacts of the trichloroethylene on the microbial community were evaluated by determining: (1) the overall microbial activity (estimated as dehydrogenase activity) and enzyme activities related to the main nutrient cycles (beta-glucosidase, o-diphenoloxidase, phosphatase, urease, and arylsulphatase activities). In addition, isoelectric focusing of the soil extracellular humic-beta-glucosidase complexes was performed to study the enzymatically active humic matter related to the soil carbon cycle. (2) The soil bacterial diversity and the molecular mechanisms for the bacterial resistance to organic solvents were also determined. For this, polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) was used to detect changes in bacterial community structure and PCR-single-strand conformational polymorphism (SSCP) was developed and optimised for detection and discrimination of the resistance-nodulation-division (RND) genes amplified from the contaminated soils. RESULTS: Vermicompost reduced, with respect to the unamended soil, about 30% of the trichloroethylene leaching during the first month of the experiment. Trichloroethylene had a marked negative effect on soil dehydrogenase, beta-glucosidase, urease, phosphatase, and arylsulphatase activities. Nevertheless, the vermicompost tended to avoid this toxic effect. Vermicompost also displays stable humic-beta-glucosidase complexes that increased the extracellular activity related to C-cycle in the contaminated soils. The isoelectric focusing technique showed a more biochemically active humic matter in the soil sampled under the vermicompost. The behaviour of the three main phyla of bacteria isolated from the DGGE bands was quite different. Bands corresponding to Actinobacteria disappeared, whereas those affiliated with Proteobacteria remained after the trichloroethylene contamination. The disappeared Actinobacteria became visible in the soil amended with the vermicompost. Bands corresponding to Bacteriodetes appeared only in columns of contaminated soils. In this study, six types of RND proteins were detected by PCR-SSCP in the natural soil, three in the trichloroethylene-contaminated soil and 7/5 in trichloroethylene-contaminated soil above/below the vermicompost in the biobarrier columns. Trichloroethylene tended to reduce or eliminate all the clones detected in the uncontaminated soil, whereas new efflux pumps appeared in the biobarrier columns. DISCUSSION: Although enzymes incorporated into the humic substances of vermicomposted olive wastes are quite stable, trichloroethylene also inhibited the background levels of the soil extracellular beta-glucosidase activity in the amended soils. The decrease was less severe in the biobarrier system, but in any case, no relation was found between the levels of trichloroethylene in soil and extracellular beta-glucosidase activity, or between the latter and the quantity of humic carbon in soils. The isoelectric focusing technique was carried out in the humic fraction to determine whether the loss of activity occurred in overall extracellular beta-glucosidase or in that linked to stable humic substances (humic-enzyme complexes). The contaminated soils showed the lower enzyme activities, whereas contaminated and amended soils presented greater quantity of focalised (and therefore stable) humic carbon and spectra heterogeneity: very different bands with higher enzyme activities. No clear relationship between trichloroethylene concentration in soil and diversity of the bacterial population was noted. Similar patterns could be found when the community structures of bacteria and microbial activity were considered. Since the use of the dehydrogenase assay has been recognised as a useful indicator of the overall measure of the intensity of microbial metabolism, these results could be attributed to PCR-DGGE methodology, since the method reveals the presence of dominant populations regardless of their metabolic state. Trichloroethylene maintained or even increased the number of clones with the DNA encoding for RND proteins, except for the contaminated soil located above the vermicompost. However, the main effect of trichloroethylene was to modify the structure of the community in contaminated soils, considering the type of efflux pumps encoded by the DNA extracted from soil bacteria. CONCLUSIONS: Trichloroethylene inhibited specific functions in soil and had a clear influence on the structure of the autochthonous bacterial community. The organic matter released by the vermicomposted olive waste tended to avoid the toxic effect of the contaminant. Trichloroethylene also inhibited the background levels of the soil extracellular beta-glucosidase activity, even when vermicompost was present. In this case, the effect of the vermicompost was to provide and/or to stimulate the humic-beta-glucosidase complexes located in the soil humic fraction >10(4), increasing the resistance of the enzyme to the inhibition. The bacterial community from the soil presented significantly different mechanisms to resistance to solvents (RND proteins) under trichloroethylene conditions. The effect of the vermicompost was to induce these mechanisms in the autochthonous bacterial community and/or incorporated new bacterial species, able to grow in a trichloroethylene-contaminated ambient. Coupled biochemical and molecular methodologies are therefore helpful approaches in assessing the effect of an organic amendment on the biochemical and biological restoration of a trichloroethylene-contaminated soil. RECOMMENDATIONS AND PERSPECTIVES: Since the main biochemical and biological effects of the organic amendment on the contaminated soil seem to be the incorporation of biochemically active humic matter, as well as new bacterial species able to grow in a trichloroethylene-contaminated ambient, isoelectric focusing and PCR-SSCP methodologies should be considered as parts of an integrated approach to determine the success of a restoration scheme.

Metabolic and bacterial diversity in soils historically contaminated by heavy metals and hydrocarbons.

The aim of this study was to characterize soils contaminated by different levels of heavy metals and hydrocarbons (Madonna Dell'Acqua, Pisa, Italy). The soils were chemically and biochemically analysed by measuring the standard chemical properties and some enzyme activities related to microbial activity (dehydrogenase activity) and the soil carbon cycle (total and extracellular beta-glucosidase activities). The metabolic capacities of soil microorganisms to degrade hydrocarbons through catechol 2,3-dioxygenase were also described. The microbial diversity of contaminated and uncontaminated soils was estimated by denaturing gradient gel electrophoresis (DGGE) of amplified 16S rDNA sequences. The PCR/single-strand conformation polymorphism (PCR/SSCP) method was used to estimate the genetic diversity of PAH-degrading genes in both contaminated and uncontaminated soils. A greater bacterial diversity and lower catechol 2,3-dioxygenase activity was detected in unpolluted soils. The complexity of the microbial community (Shannon and Simpson indices) as well as the dehydrogenase soil activity negatively correlated with contamination levels. The greatest PAH-degrading gene diversity and the most intense catechol 2,3-dioxygenase activity were found in the soils with the highest levels of hydrocarbons. Heavy metals and hydrocarbon pollution has caused a genetic and metabolic alteration in microbial communities, corresponding to a reduction in microbial activity. A multi-technique approach combining traditional biochemical methods with molecular-based techniques, along with some methodological improvements, may represent an important tool to expand our knowledge of the role of microbial diversity in contaminated soil.

Potential use of olive-waste ash from cogeneration plants as a soil amendment.

A greenhouse study was conducted to evaluate the potential use of olive-cake ash as a soil amendment, using pepper (Capsicum annuum, L. cv Italian sweet). Three soils of different pH (acidic, neutral and calcareous) were used. Treatments included a control (no fertilizer application), NPK fertilizer, and two ash-application rates that provided a complete dose (equivalent to the K2O amount in the fertilizer) and a half dose (equivalent to half the K2O amount in the fertilizer), respectively. The ash was effective in raising soil pH. Ash treatments increased the pepper (stems and leaves) dry matter yield over control; although these increases were lower than treatment including NPK. Application of ash significantly increased leaf P concentration and AB-DTPA extractable P in soil, especially in the acidic and neutral soils. Leaf K concentrations and readily and slowly available K forms in soils were affected positively by the addition of the ash. These results demonstrate that ash from the combustion of wet olive cake can be used as a beneficial organic soil amendment.

Development and assessment of an innovative soil-washing process based on the use of cholic acid-derivatives as pollutant-mobilizing agents.

Surfactant-aided soil washing is often proposed for the restoration of aged organic pollutant-contaminated soils. As many of commercial surfactants have been found to be toxic and recalcitrant, the opportunity to use in this process cheap, non-toxic, and biodegradable pollutant-mobilizing agents, such as deoxycholic acid (DA), bovine bile (BB), and the residue resulting from DA extraction from BB (BBR), was studied in this work. A soil historically contaminated by chlorinated anilines and benzenes, thiophenes, and several polycyclic aromatic hydrocarbons was suspended at 15% w/v and washed in water or water amended at 1.0% (w/v) with DA, BB, BBR, or Triton X-100 (TX). The resulting effluents were supplemented with nutrients and subjected to aerobic bioremediation. The biogenic agents enhanced the water pollutant elution potential by 230/440%. TX enhanced the same parameter by about 540%; however, it mediated a lower depletion of the initial soil ecotoxicity and a more extensive mobilization of soil constituents with respect to the biogenic agents. Furthermore, TX adversely affected the biotreatability of resulting effluents, by adversely affecting the growth of cultivable bacterial biomass and the structure of eubacterial community of the effluent. On the contrary, the biogenic agents, and in particular DA and BB, enhanced the effluents bioremediation, by sustaining the growth and increasing the complexity of the effluent eubacterial communities. Thus, DA and BB are very promising additives for an effective and environmental friendly soil washing treatment of aged (chloro)organics contaminated soils.

Vermicomposting of winery wastes: a laboratory study.

In Mediterranean countries, millions of tons of wastes from viticulture and winery industries are produced every year. This study describes the ability of the earthworm Eisenia andrei to compost different winery wastes (spent grape marc, vinasse biosolids, lees cakes, and vine shoots) into valuable agricultural products. The evolution of earthworm biomass and enzyme activities was tracked for 16 weeks of vermicomposting, on a laboratory scale. Increases in earthworm biomass for all winery wastes proved lower than in manure. Changes in hydrolytic enzymes and overall microbial activities during the vermicomposting process indicated the biodegradation of the winery wastes. Vermicomposting improved the agronomic value of the winery wastes by reducing the C:N ratio, conductivity and phytotoxicity, while increasing the humic materials, nutrient contents, and pH in all cases. Thus, winery wastes show potential as raw substrates in vermicomposting, although further research is needed to evaluate the feasibility of such wastes in large-scale vermicomposting systems.

Complicaciones de las tecnicas endovasculares en el tratamiento de aneurismas cerebrales / Endovascular technical complications in the treatment of cerebral aneurysms

Objective: To describe the complications of endovascular surgery and the morbimortality rates. Methods: 112 patients with 120 sacular aneurysms were treated by endovascular coiling approach. Technical complications related were observed in 8 patients (7.1 por ciento. The complications were mainly hemorragic and tromboembolic. Tromboembolic events were more frequent than hemorraghic events. We described some of our complications: isquemic and aneurysmal ruptures. Results: patients with tromboembolic complications has 0 por ciento mortality and 0.8 por ciento morbidity. The mortality in patients with hemorraghic complications was 1.8 por ciento and morbidity 0 por ciento. Conclusion: our experience in endovascular approach to sacular aneurysms indicates that the morbi-mortality is aceptable. This surgical technique is not free of complications