The effect of pretreatment on anaerobic activity of olive mill wastewater using batch and continuous systems

This study was focused on several physico-chemical and biological treatment methods that may affect the reduction of the organic load in olive mill wastewater (OMW). In this study, removal of 95% of the phenolic compounds present in OMW was achieved using sand filtration and subsequent treatment with powdered activated carbon in a batch system. This pretreatment for OMW was found to enhance the anaerobic activity of the sludge in the batch system significantly. The efficiency of organic load removal achieved by the anaerobic treatment of untreated OMW in batch reactors with tap water dilution factors below 1:10, reached approximately 65% chemical oxygen demand (COD) removal. However, in the up-flow sludge anaerobic blanket (UASB) reactor, COD removal efficiency of 80–85% was reached at a hydraulic retention time (HRT) of 5 days with an influent COD concentration of 40 g l⁻¹ and organic loading rate (OLR)=8 g⁻¹ COD l⁻¹ per day.     

Modelling of the mesophilic anaerobic co-digestion of olive mill wastewater with olive mill solid waste using anaerobic digestion model No. 1 (ADM1)

The anaerobic digestion model No. 1 (ADM1), conceived by the international water association (IWA) task group for mathematical modelling of anaerobic digestion processes is a structured generic model which includes multiples steps describing biochemical and physicochemical processes encountered in the anaerobic degradation of complex organic substrates and a common platform for further model enhancement and validation of dynamic simulations for a variety of anaerobic processes. In this study the ADM1 model was modified and applied to simulate the mesophilic anaerobic co-digestion of olive mill wastewater (OMW) with olive mill solid waste (OMSW). The ADM1 equations were coded and implemented using the simulation software package MATLAB/Simulink. The most sensitive parameters were calibrated and validated using updated experimental data of our previous work. The results indicated that the ADM1 model could simulate with good accuracy: gas flows, methane and carbon-dioxide contents, pH and total volatile fatty acids (TVFA) concentrations of effluents for various feed concentrations digested at different hydraulic retention times (HRTs) and especially at HRTs of 36 and 24 days. Furthermore, effluent alkalinity and ammonium nitrogen were successfully predicted by the model at HRTs of 12 and 24 days for some feed concentrations.     

Influence of dilution and phase separation on the anaerobic digestion of olive mill wastewaters

The high organic content of Olive Mill Wastewaters (OMW) causes some difficulties in maintaining the anaerobic process efficiency at high level. The two phase anaerobic system was used to treat olive mill wastewaters, diluted with tap water. Phase separation was accomplished through control of the hydraulic retention time and initial COD removal in two reactors operated in series. The effect of substrate concentration and phase separation on removal efficiency has been investigated. Experimental results indicated that yield of 0.322 to 0.335 litre biogas/g COD removal were obtained with two phase anaerobic treatment and space loading rate of 2.3 and 2.4 gCOD/l.day. The maximum methane production rate near to the theoretical value and corresponded to 360 ml of CH₄ for 1g COD removal.Abbreviations OMW Olive Mill Wastewaters - VFA Volatile Fatty Acid - COD Chemical Oxygen Demand - HRT High Retention Time - TKN Total Kjeldahl Nitrogen     

Performance and microbial communities of a continuous stirred tank anaerobic reactor treating two-phases olive mill solid wastes at low organic loading rates

A study of the performance and microbial communities of a continuous stirred tank reactor (CSTR) treating two-phases olive mill solid wastes (OMSW) was carried out at laboratory-scale. The reactor operated at a mesophilic temperature (35 degrees C) and an influent substrate concentration of 162 g total chemical oxygen demand (COD)L(-1) and 126 g volatile solids (VS)L(-1). The data analyzed in this work corresponded to a range of organic loading rates (OLR) of between 0.75 and 3.00 g CODL(-1)d(-1), getting removal efficiencies in the range of 97.0-95.6%. Methane production rate increased from 0.164 to 0.659 L CH(4)L(reactor)(-1)d(-1) when the OLR increased within the tested range. Methane yield coefficients were 0.225 L CH(4)g(-1) COD removed and 0.290 L CH(4)g(-1) VS removed and were virtually independent of the OLR applied. A molecular characterization of the microbial communities involved in the process was also accomplished. Molecular identification of microbial species was performed by PCR amplification of 16S ribosomal RNA genes, denaturing gradient gel electrophoresis (DGGE), cloning and sequencing. Among the predominant microorganisms in the bioreactor, the Firmicutes (mainly represented by Clostridiales) were the most abundant group, followed by the Chloroflexi and the Gamma-Proteobacteria (Pseudomonas species as the major representative). Other bacterial groups detected in the bioreactor were the Actinobacteria, Bacteroidetes and Deferribacteres. Among the Archaea, the methanogen Methanosaeta concilii was the most representative species.     

Detoxification of olive mill wastewaters by Moroccan yeast isolates

A total of 105 yeast strains were isolated from Moroccan olive oil production plants and evaluated for their ability to grow in olive oil mill wastewaters (OMW). The 9 isolates that grew best on OMW were selected for further study to evaluate their effect on removal of organic pollutants and OMW phytotoxicity (barley seed germination test). The results showed that at least four yeast isolates effectively lowered the toxicity of this effluent in addition to providing very useful materials in terms of both yeast biomass (6 g/l DW) and an irrigation fluid. This group of yeast isolates significantly reduced the concentration of total phenols (44% removal) and Chemical Oxygen Demand, COD (63% removal). The best germination rate of 80% for undiluted OMW was obtained for strain Candida holstii that also increased the pH from 4.76 to 6.75. Principal component analysis of the results obtained for the best yeast strains confirmed the importance of COD and total phenol reduction along with increase of organic nitrogen and final pH for the improvement of germination rates and phytotoxic reduction. This study has highlighted the potential of indigenous yeasts in detoxification of olive mill wastewaters.     

Use of Geotrichum candidum for olive mill wastewater treatment in submerged and static culture

The production of lignin peroxidase (LiP), manganese peroxidase (MnP) and lipases by Geotrichum candidum were performed in order to control the decolourisation and biodegradation of olive mill wastewater (OMW). Optimisation of different factors showed that dilution, carbon and ammonium concentrations significantly affected decolourisation and activities of ligniolytic peroxidases (LiP and MnP) on OMW. Moreover, addition of olive oil and agitation improved the lipase production. Batch and continuous OMW treatments in settler or bubble column bioreactors showed high COD and colour removal efficiencies of 60% and 50%, respectively. Lipolytic activity was greater in the batch bubble column whereas, LiP and MnP productions were improved in the settler. The performance of the continuous processes decreased with the decrease of hydraulic retention time (HRT). It has been shown that decolourisation and biodegradation decreased with an average of 40% and 45%, respectively, by decreasing the HRT from 4 d to 1.7 d.     

Potentials of anaerobic treatment for catalytically oxidized olive mill wastewater (OMW)

The catalytically oxidized olive mill wastewater (OMW) was subjected to continuous anaerobic treatment using two treatment schemes. The 1st step in both schemes was an up-flow anaerobic sludge blanket (UASB) reactor (2 0 l). The 2nd step was either a hybrid UASB reactor or a classical one (1 0 l, each). The 1st stage was operated at constant hydraulic retention time (HRT) of 24 h. The organic loading rate (OLR) varied from 3.4 to 4.8 kgCOD/m³ d depending on the quality of the pretreated wastewater. The results obtained indicated that, the 1st step UASB reactor achieved a COD percentage removal value of 53.9%. Corresponding total BOD₅ and TSS removal were 51.5% and 68.3%, respectively.The results obtained indicated that the hybrid UASB reactor as a 2nd step produced better quality effluent as compared to the classical one. This could be attributed to the presence of the packing curtain sponge with active biomass in the sedimentation part of hybrid UASB reactor which minimizes suspended solids washout, consequently enhancement of the efficiency of the reactor.Available data showed that a two stage system consisting of a classical and a hybrid UASB reactor operated at a total HRT of 48 h and OLR of 2.0 kgCOD/m³ d provided promising results. Removal values of COD_total, BOD_5 total, TOC, VFA, oil and grease were 83%, 84%, 81%, 93% and 81%, respectively. Based on the available data, the use of a two stage anaerobic system consisting of a classical UASB reactor followed by a hybrid UASB as a post-treatment step for catalytically oxidized OMW is recommended.     

Microbiological and physicochemical characterization of olive mill wastewaters from a continuous olive mill in Northeastern Portugal

The microbiological and physicochemical characterization of samples from the different wastewaters generated during oil extraction in a continuous olive mill was performed. The main aim was to determine which of the physicochemical parameters were the best fitted to correctly characterize these residual waters. High correlations were obtained for COD, DOC, K, P and N contents with the sampling points, allowing the distinction of olive washing waters (OWW) from olive centrifuge waters (OCW) and olive mill wastewaters (OMW). These parameters were sufficient for a rapid and less costly chemical characterization of these waters. Phenols and oil and grease contents, together with low pH and dissolved oxygen contents, and high organic loads, were the most toxic for microbial populations. Microbial characterization showed that fungi were well adapted to these stressing environmental characteristics and the reuse of OMW after aerobic treatment with microbial species isolated from the effluent is considered.     

Lipase production by Aspergillus ibericus using olive mill wastewater

Olive mill wastewater (OMW) characteristics make it a suitable resource to be used as a microbial culture media to produce value-added compounds, such as enzymes. In this work, the ability of the novel species Aspergillus ibericus to discolor OMW and produce lipase was studied. An initial screening on plates containing an OMW-based agar medium and an emulsified olive oil/rhodamine-B agar medium was employed to select the strain A. ibericus MUM 03.49. Then, experiments in conical flasks with liquid OMW-based media showed that the fungus could growth on undiluted OMW, with a chemical oxygen demand (COD) of 97 ± 2 g/L, and to produce up to 2,927 ± 54 U/L of lipase. When pure OMW was used in the media, the maximum COD and color reduction achieved were 45 and 97 %, respectively. When OMW diluted to 10 % was used, A. ibericus was able to reduce phenolic and aromatic compounds by 37 and 39 %, respectively. Additionally, lipase production was found to be promoted by the addition of mineral nutrients. When the fermentations were scaled up to a 2-L bioreactor, A. ibericus produced up to 8,319 ± 33 U/L of lipase, and the maximum COD and color reduction were 57 and 24 %, respectively.     

Effect of bioaugmentation of activated sludge with white-rot fungi on olive mill wastewater detoxification

AIMS: To test the potential use of Phanerochaete chrysosporium and other white-rot fungi to detoxify olive mill wastewaters (OMW) in the presence of a complex activated sludge. To combine the aerobic with anaerobic treatment to optimize the conversion of OMW in biogas. METHODS AND RESULTS: A 25-l air lift reactor was used to pretreat OMW by white-rot fungi. Detoxification of the OMW was monitored by size exclusion HPLC analysis, chemical oxygen demand (COD)/biological oxygen demand (BOD(5)) ratio evolution, and bioluminescence toxicity test. Anaerobic treatment of OMW was performed in a 12-l anaerobic filter reactor. Efficiency of the treatment was evaluated by organic matter removal, and biogas production. By comparison with the pretreatment by activated sludge only, the bioaugmentation with Phanerochaete chrysosporium or Trametes versicolor led to high removal of organic matter, decreased the COD/BOD(5) ratio and the toxicity. The subsequent anaerobic digestion of the OMW pretreated with activated sludge-white-rot fungi showed higher biomethanization yields than that pretreated with activated sludge only. Higher loading rates (7 g COD l(-1) day(-1)) were reached without any acidification or inhibition of biomethanization. CONCLUSIONS: The use of white-rot fungi, even in the presence of complex biological consortia to detoxify OMW, proved to be possible and made the anaerobic digestion of OMW for methane production feasible. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of fungi for OMW reuse and energy production could be adapted to industrial applications.     

Integrated biological process for olive mill wastewater treatment

The biological process for OMW treatment is based on an aerobic detoxification step followed by methanization step and aerobic post-treatment.The first aerobic detoxification step of OMW supplemented with sulfate and ammonium was carried out by the growth of Aspergillus niger in a bubble column. This step decreased OMW toxicity and increased its biodegradability because of phenolic compounds degradation. Growth of A. niger resulted in 58% COD removal, with production of biomass containing 30% proteins (w/w). Filtration of OMW was enhanced by this fermentation because the suspended solids were trapped in the mycelium. The filtrate liquid was then methanized using an anaerobic filter packed with flocoor. This reactor showed a short start up and a good stability. COD removal was around 60% and the methane yield (1 CH₄/g COD removed) was close to the theoretical yield.The anaerobic filter effluent was treated in an activated sludge fluidized reactor containing olive husk as a packing material. Husks were maintained in fluidization state by the aeration. This step induces COD removal at 45% and sludge (up to 2 g/dm³).The entire process allowed a global COD reduction up to 90%; however, the black colour due to polyphenolic compounds with high molecular weight persisted.     

Phenolic removal in olive oil mill wastewater using loofah-immobilized Phanerochaete chrysosporium

Summary Olive oil mill wastewater (OMW) has a high organic load, and this is a serious concern of the olive industry. Conventional biological wastewater treatments, despite their simplicity and suitable performance are ineffective for OMW treatment since phenolics possess antimicrobial activity. In order to carry out a proper treatment of OMW, use of a microorganism able to degrade the phenolics is thus necessary. In this study the ability of Phanerochaete chrysosporium to degrade the phenolic compounds of OMW and to decrease the chemical oxygen demand (COD) using cells immobilized on loofah was examined. The basal mineral salt solution along with glucose, ammonium sulfate and yeast extract was used to dilute the OMW appropriately. The fungus did not grow on the concentrated OMW. The extent of removal in this bio-treatment, of total phenols (TP) and the COD were 90 and 50%, respectively, while the color and aromaticity decreased by 60 and 95%, respectively. The kinetic behavior of the loofah-immobilized fungus was found to follow the Monod equation. The maximum growth rate μ_max was 0.045 h⁻¹ while the Monod constant based on the consumed TP and COD were (mg/l) 370 and 6900, respectively.     

Influence of salts and phenolic compounds on olive mill wastewater detoxification using superabsorbent polymers

For a selection of nine commercially available superabsorbent polymers, the absorption capacity was evaluated for the principal absorption-inhibition constituent of OMW, mineral salts and for phytotoxic-components, the phenolic compounds. A double exponential model was established for electrical conductivities ranging 4.2-25,000 microS cm(-1). For solutions of phenolic compounds ranging 0-0.5 g l(-1), a distribution coefficient near unit was achieved, while for OMW, the phenolic compounds were concentrated inside the gel as the distribution coefficient was 1.4. Correction of OMW pH towards neutrality was found to increase the absorption capacity by up to 35%. The phytotoxicity was assessed by the germination of Lepidium sativum. Inhibition in plant growth occurred for all OMW dilutions without superabsorbent polymers application. For 5% of OMW (COD 5 gl(-1) and 200 ppm of phenolic compounds) immobilised in PNa2 (1 gl(-1)), plant growth was promoted being observed a 120% growth germination, thus indicating that olive mill wastewater detoxification occurred.     

The effect of olive mill wastewaters variability on xanthan production

AIMS: Xanthan production by Xanthomonas campestris from several olive mill wastewaters (OMW) was investigated. METHODS AND RESULTS: Maximum xanthan production of 4 g l(-1) was obtained in media with 50% OMW as sole source of nutrients. OMW storage decreased effluent quality for xanthan production. The range of effluent concentration for X. campestris growth and xanthan production varied depending on OMW extraction METHOD: Wastewaters from press and two-phase extraction methods required higher dilution rates (< 10%) than those from the three-phase extraction method (50%). Nitrogen supplementation improved xanthan production in press and two-phase OMW. CONCLUSION: Factors affecting wastewaters composition, namely, waste storage, time of olive harvesting, and method for oil extraction, were found to influence xanthan production in shake-flask cultures. SIGNIFICANCE AND IMPACT OF THE STUDY: Conditions for xanthan production from OMW should be optimized in accordance with the nature of the waste material.     

Performance evaluation of a mesophilic anaerobic fluidized-bed reactor treating wastewater derived from the production of proteins from extracted sunflower flour

A study of the anaerobic digestion of wastewater derived from the production of protein isolates from extracted sunflower flour was carried out in a laboratory-scale, mesophilic (35 degrees C) fluidized-bed reactor with saponite as bacterial support. Chemical oxygen demand (COD) removal efficiencies in the range of 98.3-80.0% were achieved in the reactor at organic loading rates (OLR) of between 0.6 and 9.3 g COD/I d, hydraulic retention times (HRT) of between 20.0 and 1.1 d and average feed COD concentration of 10.6 g/l. Eighty percent of feed COD could be removed up to OLR of 9.3 g COD/l d. The yield coefficient of methane production was 0.33 l of methane (at STP) per gram of COD removed and was virtually independent of the OLR applied. Because the buffering capacity of the experimental system was maintained at favorable levels with excess total alkalinity present at all loadings, the rate of methanogenesis was not affected by loading. The experimental data indicated that a total alkalinity in the range of 2,000-2,460 mg/l as CaCO3 was sufficient to prevent the pH from dropping to below 7.0 for OLR of up to 9.3 g COD/l d. The volatile fatty acid (VFA) levels and the VFA/alkalinity ratio were lower than the suggested limits for digester failure (0.3-0.4) for OLR and HRT up to 9.3 g COD/l d and 1.1 d, respectively. For a HRT of 0.87 d (OLR of 12.1 g COD/l d) the start of acidification was observed in the reactor.     

Biological treatment of olive mill wastewater by non-conventional yeasts

The ability of lipolytic yeasts to grow on olive mill wastewater (OMW)-based medium and to produce high-value compounds while degrading this waste, was tested. OMW collected from three-phase olive mills from the North region of Portugal were characterized and used. OMW with COD ranging from 100 g L⁻¹ to 200 g L⁻¹ were supplemented with yeast extract and ammonium chloride. Studies of OMW consumption were carried out in batch cultures of Candida rugosa, Candida cylindracea and Yarrowia lipolytica. All strains were able to grow in the OMW-based media, without dilution, to consume reducing sugars and to reduce COD. C. cylindracea was the best strain concerning the lipase production and the reduction of phenolic compounds and COD. For all strains, the phenols degradation was quite difficult, mostly when more easily degradable carbon source is still present in the medium. Among the phenolic compounds tested catechol is the most inhibitory to the cells.     

Mathematical modelling of the aerobic degradation of two-phase olive mill effluents in a batch reactor

A laboratory-scale study was conducted on the aerobic degradation of two-phase olive mill effluents (TPOME) made up of the mixture of the washwaters derived from the initial cleansing of the olives and those obtained in the washing and purification of virgin olive oil. The process was carried out in a 1-l working volume stirred tank reactor operating in batch mode at room temperature (25 °C). The reactor was operated at influent substrate concentrations of 2.80 g COD/l (TPOME 25%), 5.45 g COD/l (TPOME 50%), 8.18 g COD/l (TPOME 75%) and 10.90 g COD/l (TPOME 100%). After five days of operation time, total and soluble COD removal efficiencies of 64.3% and 66.6% were achieved respectively for the most concentrated influent used (TPOME 100%). A simplified kinetic model for studying the hydrolysis of insoluble organic matter, oxidation of soluble substrate and biomass production was proposed on the basis of the experimental results obtained. The following kinetic constants with their standard deviations were obtained for the above stages in the case of the most concentrated influent used (TPOME 100%): k₁ (kinetic constant for hydrolysis of suspended organic matter): 0.11 ± 0.01 l/(g VSS day); k₂ (kinetic constant for total consumption of soluble substrate): 0.30 ± 0.02 l/(g VSS day); k₃ (endogenous metabolism constant): 0.07 ± 0.01 per day). Finally, the biomass yield coefficient was found to be 0.30 g VSS/g COD_removed. The values of non-biodegradable total and soluble CODs obtained from the model were found to be 3 and 2 g/l, respectively. The kinetic constants obtained and the proposed equations were used to simulate the aerobic degradation process of TPOME and to obtain the theoretical values of non-soluble and soluble CODs and biomass concentration. The small deviations obtained (equal or lower than 10%) between the theoretical and experimental values suggest that the parameters obtained represent and predict the activity of the microorganisms involved in the overall aerobic degradation process of this wastewater.     

Optimization of the fermentation of olive mill waste-waters by Aspergillus niger

Summary The fermentation of olive mill waste-waters (OMW) by Aspergillus niger was studied. On the basis of factorial design experiments, suspended solids and concentration of OMW, nitrogen source, sulphate and size of inocula were all found to be significant by affecting mycelium growth and chemical oxygen demand (COD) removal. Neither the absence of yeast extract, magnesium, sodium, potassium nor of calcium limited the growth of A. niger. With media lacking additional nitrogen and sulphate, the growth was limited. The optimal inoculum obtained was between 10⁶ and 10⁷ spores/g COD. The highest biomass and the greatest COD removal were obtained with removed COD to N:SO _inf4 ^sup¨- ratios averaging 100 to 3:1.5. Offprint requests to: M. Hamdi     

Use of Yarrowia lipolytica strains for the treatment of olive mill wastewater

The principal aim of this research was to evaluate the ability of different Yarrowia lipolytica strains, having different origin, to grow in olive mill wastewater (OMW) and reduce its COD level. All the strains were able to grow in undiluted OMW; the comparison between the data obtained in a semi-synthetic medium and in OMW suggests that lipases with different specificity can be produced in relation to the medium composition. Under the adopted conditions, the reduction of the OMW COD values varied from 1.47% and 41.22% of the initial value. Some strains determined a significant reduction of polyphenol content, while other ones caused its apparent increase. Moreover, some Y. lipolytica strains, isolated from chilled foods, produced the highest citric acid concentrations. These results evidenced that some Y. lipolytica strains are good candidates for the reduction of the pollution potential of OMW and for the production of enzymes and metabolites such as lipase and citric acid.     

Activity and elution profile of laccase during biological decolorization and dephenolization of olive mill wastewater

The performance and enzymatic strategy exhibited by basidiomycete Euc-1, a laccase producing strain, was investigated during the biodegradation of olive mill wastewater (OMW). This strain yielded better decolorization of solidified OMW than Phanerochaete chrysosporium and removed 90% of phenols (initial concentration=800 mg l(-1)), 73% of color (initial A465=4.4), and 45% of chemical oxygen demand in batch cultures containing OMW. Since partial phenol removal occurred before the detection of enzymatic activity, no plausible correlation could be established between them. In contrast, decolorization occurred only after the detection of laccase activity and coincided with its production over time. Two laccase fractions (Lac1 and Lac2) were separated by chromatography. OMW strongly induced Lac2 that was almost absent in defined liquid medium. Furthermore, Lac2 was the main laccase fraction in the presence of OMW. This study pointed out that basidiomycete Euc-1 and its ligninolytic system could be a useful tool for the bioremediation of wastewater generated in the process of olive oil extraction.