Dose and frequency dependent effects of olive mill wastewater treatment on the chemical and microbial properties of soil

Olive mill wastewater (OMW) is a problematic by-product of olive oil production. While its high organic load and polyphenol concentrations are associated with troublesome environmental effects, its rich mineral and organic matter contents represent valuable nutrients. This study aimed to investigate the valorization of this waste biomass as a potential soil conditioner and fertilizer in agriculture. OMW was assayed at three doses 50, 100, and 200 m³ ha⁻¹ year⁻¹) over three successive years in olive fields. The effects of the effluent on the physico-chemical and microbial properties of soil-layers were assessed. The findings revealed that the pH of the soil decreased but electrical conductivity and organic matter, total nitrogen, sodium, and potassium soil contents increased in proportion with OMW concentration and frequency of application. While no variations were observed in phosphorus content, slow increases were recorded in calcium and magnesium soil contents. Compared to their control soil counterparts, aerobic bacteria and fungi increased in proportion with OMW spreading rates. The models expressing the correlation between progress parameters and OMW doses were fitted into a second degree polynomial model. Principal component analysis showed a strong correlation between soil mineral elements and microorganisms. These parameters were not related to phosphorus and pH.     

Enzyme and fungal treatments and a combination thereof reduce olive mill wastewater phytotoxicity on Zea mays L. seeds

The phytotoxicity of olive-mill wastewater (OMW) has been suggested to be mainly due to its phenolic components. This study investigated the impact of three different low-cost dephenolization treatments on the wastewater phytotoxicity. To this aim, germinability of maize (Zea mays L.) seeds sown on a sandy-loamy soil which had been spread with different volumes (from 40 to 160m(3)ha(-1)) of either biologically-treated OMW or relative incubation control was determined. Biological treatments included either Panus tigrinus liquid cultures or incubation with commercial laccase (1UIml(-1)) or an innovative sequential combination of laccase and P. tigrinus cultures. All treatments markedly reduced phytotoxicity and promising results were obtained with commercial laccase. In fact, germinability and mean germination times in soil spread with laccase-treated OMW, did not significantly differ from those observed in soil irrigated with tap water (control) up to OMW volumes of 120m(3)ha(-1). Although the highest phenol reduction (ca. 81%) was obtained by the sequential use of laccase and P. tigrinus, the feasibility of the enzyme treatment is undoubtedly more convincing under the technological point of view.     

Dephenolization and detoxification of olive-mill wastewater (OMW) by purified biotic and abiotic oxidative catalysts

The capability of two oxidative catalysts, a laccase from Rhus vernicifera and birnessite, a manganese oxide, in the dephenolization and detoxification of two olive-mill wastewater (OMW) samples, C1 and C2, differing for complexity and composition, was evaluated. OMW phenolic extracts (EC1 and EC2) and mono-substrate solutions of phenols mostly present in OMW samples were also tested. Birnessite was more effective than laccase in removing the phenolic content from mono-substrate solutions (more than 70% of each initial phenolic concentration) and of either OMW samples or EC1 and EC2 extracts. For instance, 60% of the total phenolic content of EC1 was removed after 48-h treatment with 5 mg mL(-1) birnessite and the efficiency was lower as greater was the complexity of the OMW sample (only 17% removal from EC2 over the same time span). Phytotoxicity tests with Lepidium sativum and Lycopersicon esculentum seeds and antibacterial toxicity tests with Bacillus megaterium were performed on crude OMW samples and their extract and exhausted fractions before and after the catalytic treatment. Results demonstrated that (a) monomeric phenols were certainly but not exclusively responsible of OMW phytotoxicity, whereas their removal led to a quite complete elimination of the toxicity toward bacterial growth; (b) other components not removable by the oxidative catalysts very likely contribute to OMW phytotoxicity; and (c) the choice of the vegetal species to use in toxicity tests might be crucial for correct and easily interpretable results. Overall the results provided useful information on the possible use of oxidative catalysts for the efficient treatment of complex aqueous wastes such as those deriving from olive industry.     

Reduction of the phenolic components in olive-mill wastewater by an enzymatic treatment and its impact on durum wheat (Triticum durum Desf.) germinability

Olive-mill wastewater (OMW), an effluent of olive oil extraction process, is annually produced in huge amounts in olive growing areas. An interesting option for its disposal is the spreading on agricultural land, provided that phytotoxic effects are neutralized. The objective of the present investigation was to evaluate the potential of an enzyme-based treatment in removing OMW phytotoxicity. To this aim, germinability experiments on durum wheat (Triticum durum Desf. cv. Duilio) were conducted in the presence of different dilutions of raw or enzyme-treated OMW. OMW treatment with laccase resulted in a 65% and 86% reduction in total phenols and ortho-diphenols respectively, due their polymerization as revealed by size-exclusion chromatography. Raw OMW exerted a significant concentration-dependent inhibition on the germinability of durum wheat seeds which was evident up to a dilution rate of 1:8. When the effluent was treated with a fungal laccase, germinability was increased by 57% at a 1:8 dilution and by 94% at a 1:2 dilution, as compared to the same dilutions using untreated OMW. The treatment with laccase also decreased the mean germination time by about 1 day as compared to untreated controls. These results show that germinability inhibition due to OMW can be reduced effectively using fungal laccase, suggesting that phenols are the main determinants of its phytotoxicity.     

Valorization of treated olive mill wastewater in fertigation practice

Olive mill wastewater (OMW) brings about a major environmental problem in Tunisia as well as in the other Mediterranean countries. Its strong organic load and its toxicity due to the presence of complex phenolic compounds have dire effects when applied to soil. To overcome this difficulty, the OMW pretreatment was investigated in the present work using the Fenton oxidation reaction with zero-valent iron. Then, this pretreated wastewater was valorized in fertigation practice. The effects of the addition of different concentrations of both treated and raw OMW on soil and cropping system were investigated. The treatment by Fenton oxidation with zero-valent iron could reduce 50 % of COD and decrease 53 % of phenolic compounds. OMW application had a temporary effect on the soil pH and EC. The results showed that the evolution of soil pH and EC was related to the organic matter of the soil which depends on the spread concentrations of raw or treated OMW. After 15-day incubation period, the soil pH and EC tended to stabilize and return to the control level. Moreover, this stabilization is faster in treated OMW than that in raw OMW especially for concentrations as high as 3 and 4 %. Plants cultivated with treated OMW showed an increase in their germination. The results pointed an improvement in the stem length of plants which is almost similar to that of the control for both pea and tomato, especially for high concentrations of 3 and 4 %.     

Lentinula edodes removes phenols from olive-mill wastewater: impact on durum wheat (Triticum durum Desf.) germinability

Olive-mill wastewater (OMW) exhibits highly phytotoxic properties, mainly due to phenols. A valuable option for OMW disposal is its agricultural use provided that phytotoxic effects are removed. The present investigation was aimed at evaluating the efficacy of the lignin-degrading fungus Lentinula edodes in achieving OMW detoxification. Germinability experiments on durum wheat showed that OMW phytotoxicity was significantly reduced by L. edodes cultures. Germinability on undiluted and twofold diluted OMW from fungal cultures was 34+/-5% and 57+/-6%, respectively, while on related incubation controls it was almost completely suppressed.These results suggest that fungal cultures of L. edodes would decrease the phytotoxicity of this waste.     

Effects of compost, pig slurry and lime on trace element solubility and toxicity in two soils differently affected by mining activities

The use of organic wastes as amendments in heavy metal-polluted soils is an ecological integrated option for their recycling. The potential use of alperujo (solid olive-mill waste) compost and pig slurry in phytoremediation strategies has been studied, evaluating their short-term effects on soil health. An aerobic incubation experiment was carried out using an acid mine spoil based soil and a low OM soil from the mining area of La Unión (Murcia, Spain). Arsenic and heavy metal solubility in amended and non-amended soils, and microbial parameters were evaluated and related to a phytotoxicity test. The organic amendments provoked an enlargement of the microbial community (compost increased biomass-C from non detected values to 35 μg g⁻¹ in the mine spoil soil, and doubled control values in the low OM soil) and an intensification of its activity (including a twofold increase in nitrification), and significantly enhanced seed germination (increased cress germination by 25% in the mine spoil soil). Organic amendments increased Zn and Pb EDTA-extractable concentrations, and raised As solubility due to the influence of factors such as pH changes, phosphate concentration, and the nature of the organic matter of the amendments. Compost, thanks to the greater persistence of its organic matter in soil, could be recommended for its use in (phyto)stabilisation strategies. However, pig slurry boosted inorganic N content and did not significantly enhance As extractability in soil, so its use could be specifically recommended in As polluted soils.     

Short-term impact of olive mill wastewater (OMWW) applications on the physico-chemical and microbiological soil properties of an olive grove in Argentina

The purpose of this work was to investigate the effects of spreading olive oil mill wastewater (OMWW) on soil biochemical parameters and olive production in an organically managed olive orchard. The experiment was carried out with three different doses of OMWW (80, 160 and 500 m³ ha^?1) and a control (untreated soil). Three samplings were done at 10, 30 and 90 days after the administration of the byproduct. OMWW application differentially modified the biochemical properties of the soil analyzed. Organic matter, organic carbon, total nitrogen and extractable phosphorus soil contents increased proportionally with each increasing dose. The values of these parameters decreased gradually with time. Total microbial activity was altered and the OMWW 500 m³ ha^?1 treatment proved to be the most active when compared with the other applied doses. OMWW agricultural application also modified the structure of soil microbial communities, particularly affecting Gram positive and negative bacteria, while fungal biomass did not show consistent changes. Although there was a salinity increase in the treated soil, especially at the highest dose, the productive parameters analyzed (fruit and oil tree^?1) were not affected. In light of the obtained results, we consider that low dose of OMWW could be considered an alternative farming practice for semiarid regions.     

The evaluation of stability and maturity during the composting of cattle manure

We examined chemical, microbiological and biochemical parameters in order to assess their effectiveness as stability and maturity indicators during the composting process of cattle manure. The composting material obtained after 15 d in trenches and at different times during the maturation phase (i.e. 80, 180 and 270 d) were analyzed. We found that the material collected at the end of the active phase was inadequate to be applied to soil as organic amendment due to its high content of NH4+, its high level of phytotoxicity and the low degree of organic matter stability. After a maturation period of 80 d, the stability of the sample increased. This was shown by a reduction in the dissolved organic carbon (DOC) content and NH4+ concentration and also by a reduction in the microbial activity and biomass; however, 180 d of composting were not sufficient to reduce the phytotoxicity to levels consistent for a safe soil application. Among the various parameters studied, the change in DOC with composting time gave a good indication of stability.     

Transformation of dissolved organic matter in a novel groundwater recharge system with reclaimed water

A novel process, enhanced direct injection-well recharge system (EnDir), can overcome the technical difficulties during the application of conventional surface spreading and has been developed to recharge groundwater with reclaimed water. In this study, removal and transformation of dissolved organic matter (DOM) in the system were investigated in laboratory-scale experiments. Results demonstrated that dissolved organic carbon and trihalomethane formation potential values could be reduced from 6.54 +/- 1.30 mg/L and 267.9 +/- 24.3 microg/L to 1.59 +/- 0.64 mg/L and 104.5 +/- 10.2 microg/L, respectively, as a result of DOM biodegradation in the aerobic short-term vadose soil treatment. Fluorescence spectra showed that aromatic protein-like substances and soluble microbial byproducts could be removed, to a great extent, in the soil system. Despite different removal efficiencies of DOM in different molecular weight fractions, the residual DOM was composed mainly of fulvic acid-like and humic acid-like substances, with molecular weights of 500 Da to 1 kDa.     

Long-Term Effects on Soil of the Disposal of Olive Mill Waste Waters (OMW)

Disposal of untreated olive mill waste waters (OMW) is a serious environmental problem in many Mediterranean countries. The aim of this work was to assess whether changes in soil properties have occurred due to direct disposal of raw OMW on soil and in evaporation ponds, and to investigate the potential fate and transport of pollutants after olive oil production had ceased. The results clearly showed that uncontrolled OMW disposal is a significant source of pollution to surface soil and waters. Disposal of OMW on soil greatly increased electrical conductivity (EC), available phosphorous (P), exchangeable potassium (K) and magnesium (Mg), organic matter, polyphenols, total and inorganic nitrogen (N), and available micronutrients mainly in surface soil layers. The presence of a high content of clay and carbonates in soil act as barriers and prevent downward transport of pollutants. Residual levels of total carbon (C), polyphenols, total and inorganic N, exchangeable K⁺, available P, iron (Fe), and copper (Cu) were also elevated even 8 years after mill closure. The long term disposal of OMW highlights the need to establish soil quality standards for soil parameters in order to identify soils affected by the disposal of OMW.     

不同人工湿地基质对污水总有机碳去除能力初探

选择4种常见基质及基质配比,研究其碳含量和组分对污水中总有机碳(TOC)的去除影响。结果显示当污水的TOC浓度为27 mg/L时,4种基质都向水体中释放碳,且释放碳的大小顺序为:砂子-土壤-泥炭混合基质土壤砂子-土壤混合基质砂子,基质的碳含量越高,释放到水体中碳越多。3种基质对水体中TOC的去除率大小顺序为:土壤砂子-土壤混合物基质砂子,基质碳含量越高,微生物活性越强,对TOC的去除率越高。在第12天,砂子-土壤-泥炭混合基质处理中TOC浓度的增加,与其稳结合态有机碳含量较高有关。有机碳含量和组分影响了人工湿地基质对TOC的去除。     

Influence of lead acetate on soil microbial biomass and community structure in two different soils with the growth of Chinese cabbage (Brassica chinensis)

A greenhouse pot experiment was conducted to evaluate the impact of different concentrations of lead acetate on soil microbial biomass and community structure during growth of Chinese cabbage (Brassica chinensis) in two different soils. The field soils were used for a small pot, short-term 60-day growth chamber study. The soils were amended with different Pb concentrations, ranging from 0 to 900mgkg(-1) soil. The experimental design was a 2 soilx2 vegetation/non-vegetationx6 treatments (Pb)x3 replicate factorial experiment. At 60 days the study was terminated and soils were analyzed for microbial parameters, namely, microbial biomass, basal respiration and PLFAs. The results indicated that the application of Pb at lower concentrations (100 and 300mgkg(-1)) as lead acetate resulted in a slight increase in soil microbial biomass, whereas Pb concentrations >500mgkg(-1) caused an immediate gradual significant decline in biomass. However, the degree of impact on soil microbial biomass and basal respiration by Pb was related to management (plant vegetation) or the contents of clay and organic matter in soils. The profiles of 21 phospholipid fatty acids (PLFAs) were used to assess whether observed changes in functional microbial parameters were accompanied by changes in the composition of the microbial communities after Pb application at 0, 300 and 900mg Pbkg(-1) soil. The results of principal component analyses (PCA) indicated that there were significant increases in fungi biomarkers of 18:3omega6c, 18:1omega9c and a decrease in cy17:0, which is an indicator of gram-negative bacteria for the high levels of Pb treatments In a word, soil microbial biomass and community structure, therefore, may be sensitive indicators reflecting environmental stress in soil-Pb-plant system. However, further studies will be needed to better understand how these changes in microbial community structure might actually impact soil microbial community function.     

Agro-industrial wastes as effective amendments for ecotoxicity reduction and soil health improvement in aided phytostabilization

Aided phytostabilization is a technology that uses metal tolerant plants and organic and/or inorganic amendments to reduce soil metal bioavailability, while improving soil health. Our objective was to determine the effects of the application of amendments [sheep manure (SHEEP), poultry litter (POULTRY), cow slurry (COW), and paper mill sludge mixed with poultry litter (PAPER)], together with the growth of a metallicolous Festuca rubra L. population, on (i) chemical and microbial indicators of soil health and (ii) soil ecotoxicity, during the aided phytostabilization of a Zn/Pb contaminated mine soil. Amendment application led to an increase in soil pH, organic matter content, and inorganic salts, resulting in a decrease in Pb and Zn CaCl₂-extractable concentrations in soil, which, in turn, contributed to lower ecotoxicity and a stimulation of plant growth and soil microbial communities. The factor most affecting the metal extractability was probably soil pH. POULTRY was the best amendment in terms of increasing plant growth, chlorophylls content, and soil microbial biomass and activity, but resulted in higher levels of phytoavailable Pb and Zn. SHEEP and PAPER were more effective at reducing metal CaCl₂-extractability and, consequently, led to lower values of metal accumulation in plant tissues, thereby reducing the risk of metals entering into the food chain. When combined with the application of organic amendments, the metallicolous F. rubra population studied here appears an excellent candidate for aided phytostabilization. Our results indicate that the application of organic amendments is essential for the short-term recovery of highly contaminated metalliferous soils during aided phytostabilization.     

Microbial biomass and activity in a Brazilian soil amended with untreated and composted textile sludge

This laboratory study examines the effect of application of untreated and composted textile sludge on microbial biomass and activity in a Brazilian soil. The soil was amended with untreated and composted sludge at rates equivalent of 6.4t ha(-1) (0.64 g per 100g of soil) and 19t ha(-1) (1.90 g per 100g of soil), respectively, and were incubated at 28 degrees C for 60 days and daily sampled for microbial activity. An additional experiment, in the same condition, was conduced for evaluation of microbial biomass and enumeration of microorganisms at 15, 30 and 60 days after incubation. The application of composted sludge increased significantly the microbial biomass and activity, and bacteria number of soil. There were not differences in the microbial activity and bacteria number among the control and untreated sludge amended soils. In conclusion, after 2 months of incubation, the effects of the two amendments on soil microorganisms were: microbial biomass, soil respiration and bacteria number were increased only in composted sludge treated soil. qCO2 and fungi number were not affected by untreated and composted sludge.     

Release of metals from homogeneous soil columns by wastewater from an agricultural industry

The effect of discharging olive mill wastewater (OMW) in soils on the release of metals previously retained by them is simulated by leaching homogeneous soil columns with a solution of the residue after passing solutions of Cu or Zn through the columns. The effect of other residues, previously added to the soil as composts in field experiments, on the behaviour observed in the laboratory is also discussed. OMW shows a strong power for releasing the metals retained. Previous addition of a compost made from olive mill sludge and plant refuse to the soil causes a significant reduction of the release of retained metal by liquid OMW. Previous addition of concentrated sugarbeet vinasse causes much less significant effects.     

Comparison of the effectiveness of composting and vermicomposting for the biological stabilization of cattle manure

Cattle manure is produced in large quantities in industrial breeding facilities and the storage and/or spreading of this waste on land may cause contamination of the atmosphere, soil and water. The aim of the present study was to evaluate the effectiveness of the active phases of composting, vermicomposting, and also a combination of composting and vermicomposting for reducing the polluting potential and for stabilizing cattle manure in the short-term. For this, the degree of decomposition as well as the microbial activity and microbial composition of the resulting products after the active phase of composting and vermicomposting were analysed. None of the treatments significantly reduced the dissolved organic carbon and dissolved organic nitrogen contents relative to the control, and therefore more time may be required for stabilization. Nevertheless, the lowest values of microbial biomass and activity corresponded to the earthworm-worked substrates, in which fungal growth was also promoted; the combined treatment (composting + vermicomposting) was the most effective in terms of stabilizing the cattle manure. Moreover, earthworms promoted the retention of nitrogen and gradual release of P, as well as a reduction in electrical conductivity, thereby producing improved substrates for agricultural use.     

Raw and fungal-treated olive-mill wastewater effects on selected parameters of lettuce (Lactuca sativa L.) growth--the role of proline

Olive-mill wastewater (OMW) constitutes a major agricultural waste stream for which disposal is associated with significant environmental repercussions. No data are available on the effects of biotreated OMW and of the protective role of exogenously provided proline on plant physiology. In the present study, OMW was administered, either raw or previously treated by the white-rot fungus Pleurotus ostreatus, with or without proline amendment, to lettuce plants growing in sterilized sand. Biotreated OMW and proline addition resulted in significant moderation of OMW adverse effects on plant biomass production and ascorbic acid content, while their synergistic action alleviated the severe negative impact on net photosynthetic rate, water use efficiency and photosynthetic activity (Fv/Fo) invoked by the effluent. Moreover, biotreated OMW supplemented with proline, moderated the decrease in chlorophylls exerted by raw OMW, but it did not contribute at restoring carotenoids content. Restoration of plant transpiration was complete when biotreated OMW was used (with or without proline); proline alone mitigated the negative impact of OMW on photosynthetic efficiency (Fv/Fm and Fv'/Fm'). It seems that key photosynthetic parameters could be exploited as suitable evaluators of wastewater-induced plant toxicity, while plant fertigation with biotreated and/or supplemented OMW could be an interesting prospect in valorizing this effluent.     

Effects of 2,4-dichlorophenol, pentachlorophenol and vegetation on microbial characteristics in a heavy metal polluted soil

The aim of this study was to evaluate the soil microbial characteristics in historically heavy-metal polluted soil, which was also affected by organic co-contaminants, 2,4-dichlorophenol or pentachlorophenol, which often occur due to the conventional use of pesticides. It was observed that the normalized microbial biomass (microbial biomass per unit soil organic C) of the contaminated soil was very low, less than 1% in both non-planted and ryegrass planted soil, and showed a decreasing trend with the treatment of organic co-contaminants. The microbial biomass and substrate-induced respiration (SIR) in the ryegrass planted soil were much larger, as compared with the non-planted soil with or without organic pollutants. The different resistant bacterial community and its physiological diversity in the rhizosphere further suggested that the effect of vegetation on microbial activity was not just a general increase in the mass or activity of pre-existing microorganisms, but rather acted selectively on microbial growth so that the relative abundance of different microbial groups in soil was changed. In sum, high concentrations of organic co-contaminants, especially pentachlorophenol (PCP), could strengthen the deterioration of microbial ecology. The adverse effect of heavy metal-organic pollutants on the soil microbial biomass and activity might be the reason for the slow degradation of PCP that has high chlorinated and high toxicity. Vegetation might be the efficient way to assist in improving and restoring the utilization of agricultural ecosystems. The beneficial microbial effect of vegetation could cause the rapid dissipation of 2,4-dichlorophenol (2,4-DCP) that has less chlorinated and less toxicity in the planted soils.     

Effects of 2,4-dichlorophenol,pentachlorophenol and vegetation on microbial characteristics in a heavy metal polluted soil

The aim of this study was to evaluate the soil microbial characteristics in historically heavy-metal polluted soil, which was also affected by organic co-contaminants, 2,4-dichlorophenol or pentachlorophenol, which often occur due to the conventional use of pesticides. It was observed that the normalized microbial biomass (microbial biomass per unit soil organic C) of the contaminated soil was very low, less than 1% in both non-planted and ryegrass planted soil, and showed a decreasing trend with the treatment of organic co-contaminants. The microbial biomass and substrate-induced respiration (SIR) in the ryegrass planted soil were much larger, as compared with the non-planted soil with or without organic pollutants. The different resistant bacterial community and its physiological diversity in the rhizosphere further suggested that the effect of vegetation on microbial activity was not just a general increase in the mass or activity of pre-existing microorganisms, but rather acted selectively on microbial growth so that the relative abundance of different microbial groups in soil was changed. In sum, high concentrations of organic co-contaminants, especially pentachlorophenol (PCP), could strengthen the deterioration of microbial ecology. The adverse effect of heavy metal-organic pollutants on the soil microbial biomass and activity might be the reason for the slow degradation of PCP that has high chlorinated and high toxicity. Vegetation might be the efficient way to assist in improving and restoring the utilization of agricultural ecosystems. The beneficial microbial effect of vegetation could cause the rapid dissipation of 2,4-dichlorophenol (2,4-DCP) that has less chlorinated and less toxicity in the planted soils.