High-Rate Anaerobic Digestion of Waste Activated Sludge by Integration of Electro-Fenton Process.

Anaerobic digestion (AD), being the most effective treatment method of waste activated sludge (WAS), allows for safe disposal. The present study deals with the electro-Fenton (EF) pretreatment for enhancing the WAS biogas potential with low-cost iron electrodes. The effect of pretreatment on the physicochemical characteristics of sludge was assessed. Following EF pretreatment, the pH, conductivity, soluble chemical oxygen demand (SCOD), and volatile fatty acids (VFA) increased to 7.5, 13.72 mS/cm, 4.1 g/L, and 925 mg/L, respectively. Capillary suction time (CST) analysis highlighted the dewaterability effect of EF on WAS, as demonstrated by the decrease in CST from 429 to 180 s following 30 min of pretreatment. Batch digestion assays presented an increase in the biogas yield to 0.135 L/g volatile solids (VS) after 60 min of EF pretreatment in comparison to raw sludge (0.08 L/g VS). Production of biogas was also found to improve during semi-continuous fermentation of EF-pretreated sludge conducted in a lab-scale reactor. In comparison to raw sludge, EF-pretreated sludge produced the highest biogas yield (0.81 L biogas/g VS) with a high COD removal rate, reaching 96.6% at an organic loading rate (OLR) of 2.5 g VS/L. d. Results revealed that the EF process could be an effective WAS disintegration method with maximum recovery of bioenergy during AD.

Protective effect of Dunaliella sp., lipid extract rich in polyunsaturated fatty acids, on hepatic and renal toxicity induced by nickel in rats.

AIMS: The present study was undertaken to investigate the protective effect of lipid extract of Dunaliella sp. (LE) rich in polyunsaturated fatty acids (PUFA), against oxidative stress induced by nickel in experimental rats. METHODS: Our investigation evaluated the antioxidant activity of LE using both DPPH and NBT assays. Twenty female albino Wistar rats, randomly allocated into four experimental groups, namely (C): control, (Nit): nickel-treated rats with 5 mg/kg/d of NiCl2 during 30 days, (LEa): lipid extract-administered rats with 5 mg/kg BW/d during 30 days and (Nit + LEa): rats treated with Ni and LE-administered during 30 days. RESULTS: The in vitro antioxidant activity demonstrated that LE presents an important antioxidant potential. In vivo, the (Nit + LEa) cotreatment decreased the level of malondialdehyde and restored the antioxidant enzyme activities (superoxide dismutase, catalase and glutathione peroxidase) in livers and kidneys in comparison with those treated with Ni only. LE administration to rats treated with Ni also ameliorated biochemical and histological parameters as compared to only Ni-treated group. LE of Dunaliella sp., rich in polyunsaturated fatty acids showed a significant hepato- and reno-protective effect against metal-induced toxicity. CONCLUSION: LE of Dunaliella sp., rich in PUFA has been proven to be effective in protection against Ni-induced toxicity.

Characterization of Amphora sp., a newly isolated diatom wild strain, potentially usable for biodiesel production.

Microalgae as feedstock for biofuel production have attracted serious consideration as an important sustainable source of energy. For biodiesel production with microalgae, a series of consecutive processes should be performed as selection of adequate microalgal strains, mass culture, cell harvesting, oil extraction and transesterification. The aim of this study was to investigate the growth and lipid accumulation of a new isolated marine microalgal strain by optimizing culture medium composition and applying different stressful culture conditions. Microalga CTM 20023 was isolated from the evaporating salt-ponds at Sfax, Tunisia, using serial-dilution technique from enriched cultures. Phylogenetic analysis based on SSU rDNA and rbcL-3P sequences attributed this isolate to a new species of the Amphora genus. This wild strain possesses rapid gravity sedimentation of 2.91 m h(-1), suitable for an easy and low-cost biomass harvest. The optimization of the composition of the culture medium through statistical experimental designs improved the specific growth rate of Amphora sp. from 0.149 to 0.262 day(-1) and increased its 15-day culture biomass production from 465 to 2200 mg L(-1) (dw) and its lipid content from 140 to 370 mg g(-1) (dw). Highest biomass productivity of 178 mg L(-1) day(-1) was achieved at the 10th day of culture. Highest lipid content of 530 mg g(-1) (dw) was obtained under phosphorus starvation and 64.34% of these lipids were saturated fatty acids. A first growth stage, in optimized condition, would thus offer the maximum productivity for an algal biomass feed stream, followed by second stressful stage for lipid accumulation, thus suitable for biodiesel production.

Optimisation of the critical medium components for better growth of Picochlorum sp. and the role of stressful environments for higher lipid production.

BACKGROUND: Coastal countries that suffer from a scarcity of water, such as Tunisia, have to cultivate marine microalgae on non-arable land in order to produce feedstock and overcome their demands of nutrition and energy. In this framework, a green microalga, CTM 20019, was isolated, identified as Picochlorum sp. and tested for its lipid production. RESULTS: The dry weight of Picochlorum sp. is composed of 163 g kg(-1) lipids, 225 g kg(-1) total sugars, 440 g kg(-1) proteins and 112 g kg(-1) ash rich in potassium, calcium, iron, magnesium and zinc. Gas chromatography-mass spectrometry analysis showed that the main fatty acids were palmitic acid (29%), linolenic acid (26.5%), linoleic acid (23.5%), hexadecatrienoic acid (11%) and hexadecadienoic acid (8.5%). As it is known that culture conditions greatly influence the composition of microalgae, the experiments were designed to optimise the composition of the medium in order to increase Picochlorum sp. growth from OD680nm = 0.53 to OD680nm = 2.2 and lipid accumulation from 163 g kg(-1) to 190 g kg(-1) . The highest lipid contents of 570 and 585 g kg(-1) were achieved under phosphate starvation and sodium carbonate supplementation, respectively. Under these conditions, the fatty acid profile is dominated by mono-unsaturated and polyunsaturated acids, and is therefore suitable for aqua-culture feeding. However, under high salinity, growth and lipid synthesis are inhibited, and the fatty acids are saturate, and the product is therefore suitable for biodiesel. CONCLUSION: This high lipid content rich in essential fatty acids, omega-6 and omega-3, endorses this wild strain of Picochlorum sp. as a promising feedstock for aqua-culture and human nutrition or for the production of biodiesel. © 2013 Society of Chemical Industry.

Enzymatic oxidative transformation of phenols by Trametes trogii laccases.

The removal of toxic phenolic compounds from industrial wastewater is an important issue to be addressed. Their presence in water and soil has become a great environmental concern, and effective methods for their removal need to be addressed. The feasibility of applying laccases for the degradation of phenolic compounds has received increasing attention. In the present work, the transformation of five phenolic compounds (catechol, hydroxytyrosol, tyrosol, guaiacol and p-coumaric acid), the main constituents of a typical wastewater derived from an olive oil factory, by Trametes trogii laccases was studied at concentrations ranging between 0.2 and 1.6 mM. High-performance liquid chromatography analysis showed high degradation rates of phenolic compounds by T trogii laccases. Independently of the used concentration, a complete transformation of guaiacol, p-coumaric acid, hydroxytyrosol and tyrosol occurred after 1 h of incubation. The transformation of catechol depends on its initial concentration. The liquid chromatography-mass spectrometry analysis showed that laccases catalysed transformation of p-coumaric acid and tyrosol, resulting in the formation of phenolic dimers. No reduction of enzyme activity has been observed during the oxidation of all phenolic compounds. These results suggest that the studied laccases were capable of efficiently removing phenolic compounds, as well as catalysing the production of novel phenolic dimers.

Isolation, identification and characterization of a new lipolytic pseudomonas sp., strain AHD-1, from Tunisian soil.

A novel, lipid-degrading bacterium (strain AHD-1) was isolated from soil regularly contaminated with washing-machine wastewater in Sfax, Tunisia. When this strain was grown in a medium containing 2% triacylglycerol, the hydrolysis products were found to be diacylglycerols, monoacylglycerols and free fatty acids. This strain was an aerobic, mesophilic, Gram-negative, motile, non-sporulating bacterium, capable of growing optimally at pH 7 and 27 degrees C. The predominant fatty acids were found to be C16:1omega7c (31%), C16:0 (28.1%), C18:1 omega7c (16.3%) and C17:0 (5.8%). Phylogenetic analysis of the 16S rRNA gene showed that this isolate is a new strain belonging to the genus Pseudomonas. Strain AHD-1 was found to be closely related to Pseudomonas azotoformans IAM 1603T, Pseudomonas gessardii CIP 105469T and Pseudomonas libanensis CIP 105460T with 99.7%, 99.56% and 99.54% of similarity, respectively.

Comparison between Thermo-Alkaline and Electro-Fenton Disintegration Effect on Waste Activated Sludge Anaerobic Digestion.

Disintegration of municipal waste activated sludge (WAS) using thermo-alkaline (TA) and electro-Fenton (EF) methods was investigated and compared in terms of the efficiency of sludge solubilisation and enhancement of anaerobic biodegradability. Performance of organic matter solubilisation (soluble COD, proteins, polysaccharides) of sludge pretreated with EF was proved to be better than that with TA pretreatment, which resulted in the enhancement of anaerobic biodegradability. Comparison of results indicated that percentages of PN and PS release obtained after EF pretreatment (68.95 and 65.22%) were higher than those obtained by TA method (45.25 and 35.22%) respectively. An improvement of biogas potential about 2 and 1.6 times was achieved respectively by EF and TA pretreatment in comparison to raw sludge. During semi-continuous fermentation study in continuous stirred tank reactor, EF pretreated sludge gave the best biogas yield (0.6 L biogas/g COD) at an OLR of 2.5 g COD/L. d in comparison to TA pretreated sludge (0.3 L biogas/g COD), where low biogas yield about 0.1 L biogas/g COD was registered by raw sludge in the same CSTR. Therefore, the integration of EF process to anaerobic digestion might be a promising process for sludge reduction and biogas recovery.

Nitrogen or phosphorus repletion strategies for enhancing lipid or carotenoid production from Tetraselmis marina.

The objective of this study was to investigate the accumulation of lipid and photosynthetic pigments from Tetraselmis marina. When the cells were grown in F/2-medium for seven days in the first stage, the carotenoid and lipid contents, and productivity were 44g/kg (DW), 27% and 31mg/L/d, respectively. After second stage of cultivation of T. marina for further 3-days under N-replete condition (4.41mM NaNO3) increased biomass concentration of 1900mg/L and lipid content of 50% were observed, with an enhanced lipid productivity of 86.36mg/L/d and SFA and MUFA fractions of 70.76 and 13.14%, respectively. However, under P-repletion (2.08mM NaH2PO4), its carotenoid content increased to 89.23g/kg and its PUFA for 65% of total lipids. Results showed that N and P-replete conditions decreased SOD activity and increased H2O2 and TBARS levels of T. marina. Thus, this native microalga strain could be a potent candidate for feed,  food or biofuel production.

Changes in microbial and soil properties following amendment with treated and untreated olive mill wastewater.

We investigated the effect of untreated and biologically treated olive mill wastewater (OMW) spreading on the soil characteristics and the microbial communities. The water holding capacity, the salinity and the content of total organic carbon, humus, total nitrogen, phosphate and potassium increased when the spread amounts of the treated or untreated OMW increased. The OMW treated soil exhibited significantly higher respiration compared to the control soil. However, the C-CO2/C(tot) ratio decreased from 1.7 in the control soil to 0.5 in the soil amended with 100 m3 ha(-1) of untreated OMW. However, it slightly decreased to 1.15 in the soil amended with 400 m3 ha(-1) of treated OMW. The treated OMW increased the total mesophylic number while the number of fungi and nitrifiers decreased. Actinomycetes and spore-forming bacteria were neither sensitive to treated nor to untreated OMW. The total coliforms increased with higher doses of treated and untreated OMW. A toxic effect of the untreated OMW appeared from 100 m3 ha(-1). This toxicity was more significant with 200 m3 ha(-1), where microflora of total mesophilic, yeasts and moulds, actinomycetes, and nitrifiers were seriously inhibited except for total coliforms and spore-forming bacteria.

Salinity stress increases lipid, secondary metabolites and enzyme activity in Amphora subtropica and Dunaliella sp. for biodiesel production.

Amphora subtropica and Dunaliella sp. isolated from Tunisian biotopes were retained for their high lipid contents. Respective optimized parameters for rapid growth were: pH 9 and 10, light period 21 and 24h and temperature 31 and 34°C, respectively. After optimization, Amphora subtropica growth rate increased from 0.2 to 0.5day(-1) and Dunaliella sp. growth rate increased from 0.38 to 0.7day(-1). Amphora subtropica biomass production, productivity and lipid content increased from 0.3 to 0.7gL(-1)(dw), 69-100mgL(-1)d(-1)(dw) and 150-190gkg(-1)(dw), respectively, and Dunaliella sp. from 0.5 to 1.4gL(-1)(dw), 124-200mgL(-1)d(-1) (dw) and 190-280gkg(-1)(dw), respectively. Often to overcome trade-off between microalgae rapid growth and high lipid content which are often conflicting and very difficult to obtain at the same time, separation in a growth stage and a lipid accumulation stage is obvious. Salinity stress in a single stage of culture was studied. Compared to the optimal concentration of growth, excess or deficiency of NaCl engendered the same cellular responses by implication of oxidative stress systems and reactivation of defense and storage systems. Indeed, increasing salinity from 1M to 2M for Amphora subtropica or decreasing salinity from 3M to 2M for Dunaliella sp. have both increased lipids content from (220 and 280) to (350 and 430)gkg(-1), carotenoids from (1.8 and 2.4) to (2.3 and 3.7)pgcell(-1), TBARS amount from (10.4 and 5.3) to (12.1 and 10.7)nmolmg(-1) proteins and SOD activity from of (46.6 and 61.8) to (71.6 and 79.4)Umg(-1) proteins, respectively. With further improved fatty acids profile, the microalgae strains could be potent candidates for biofuel production.

Biochemical and molecular characterization of Pseudomonas aeruginosa CTM50182 organic solvent-stable elastase.

An extracellular alkaline elastase was produced from Pseudomonas aeruginosa CTM50182. It was chromatographically purified using HPLC and Mono Q Sepharose column. Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis revealed that the purified enzyme (called AMPP) was a monomer with a molecular mass of 33,015.18 Da. The N-terminal 29 amino acid sequence of AMPP showed high homology with those of Pseudomonas elastases. It showed optimal activity at pH 12 and 80 °C and was stable at a pH range of 9-12 after 120 h of incubation. Its thermoactivity and thermostability were upgraded in the presence of 5 mM Co(2+). Its half-life times at 70 and 80 °C were 16 and 10 h, respectively. It was completely inhibited by ethylene glycol-bis (ß-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), and 1,10-phenanthroline, suggesting that it belongs to the metalloprotease family. AMPP also exhibited high catalytic efficiency, organic solvent-tolerance, and hydrolysis. The lasB gene encoding AMPP was cloned, sequenced, and expressed in Escherichia coli. The biochemical properties of the extracellular purified recombinant enzyme (rAMPP) were similar to those of native AMPP. This organic solvent-stable protease could be considered a potential candidate for application as a biocatalyst in the synthesis of enzymatic peptides.

Effect of natural mediators on the stability of Trametes trogii laccase during the decolourization of textile wastewaters.

The purpose of the present study was to determine the effect of natural mediators on the stability of the Trametes trogii crude laccase in the process of decolourization of textile effluents. Acetosyringone allowed the highest wastewaters decolourization rate of 25%. At higher concentrations of acetosyringone, the relative activity of laccase decreased approximately by between 38% and 88% after 5 days of incubation. T. trogii laccase was strongly inactivated at 3 mM syringaldehyde, after 3 days of incubation. However, laccase activity is more stable in the presence of the vanillin and m-coumarate. The T. trogii growth on solid effluent-based-medium was examined and evaluated by measuring the colony diameter in cm. T. trogii was completely inhibited on 100:0 and 80:20 effluent:water solid medium, however, colony diameter reached 5 cm on 60:40 effluent:water solid medium after 13-14 days incubation. When the textile effluent was pre-treated with laccase and laccase-acetosyringone system, the colony diameter of 2 cm of T. trogii on 80:20 effluent:water solid medium was reached after 14 and 10 days of incubation respectively.

Decolourization and detoxification of textile industry wastewater by the laccase-mediator system.

Decolourization and detoxification of a textile industry effluent by laccase from Trametes trogii in the presence and the absence of laccase mediators was investigated. Laccase alone was not able to decolourize the effluent efficiently even at the highest enzyme concentration tested: less than 10% decolourization was obtained with 9 U/mL reaction mixture. To enhance effluent decolourization, several potential laccase mediators were tested at concentrations ranging from 0 to 1mM. Most potential mediators enhanced decolourization of the effluent, with 1-hydroxybenzotriazol (HBT) being the most effective. The effect of several physico-chemical parameters that could influence enzyme activity, such as pH, temperature and dye concentration was tested. Optimal decolourization occurred with 20% effluent at pH 5, a temperature of 50 degrees C, and in the presence of 1mM HBT. The toxicities of crude, laccase-HBT treated and laccase-acetosyringone treated effluent were evaluated using the Microtox assay. Only laccase-acetosyringone treated effluent was not toxic; crude and laccase-HBT treated effluent retained toxicity.

Assessment of the impact of excessive chemical additions to municipal wastewaters and comparison of three technologies in the removal performance of pathogens and toxicity.

The removal efficiencies of pathogens such as Salmonella (S), helminths ova (H), protozoan cysts (P), total coliforms (TC), faecal coliforms (FC) and faecal streptococci (FS) by three treatment processes: aerated lagoon (AL), activated sludge (AS) and anaerobic membrane bioreactor (MBR) were evaluated by means of standard microbiological numeration methods. The micro-toxicity and phyto-toxicity of wastewaters were monitored by LUMIStox and germination index (GI) of Lepidium sativum tests. The results of municipal wastewaters receiving industrial effluents such as Sfax and Mahres were compared with other municipal wastewaters receiving mainly domestic effluents such as Ksour-Essaf. The anaerobic MBR allowed an effective removal of 100% for all the microorganisms tested. The average removal of TC, FC, FS, S, H and P was 1.65log(10), 1.42log(10), 1.23log(10), 0.91log(10), 52.23% and 76.15% in AL system and 0.62-0.84log(10), 0.87-0.93log(10), 0.71-0.78log(10), 0.81-2.71log(10), 59-74.1% and 59.84-72.2% in AS processes, respectively. LUMIStox and GI of L. sativum tests demonstrated that Ksour-Essaf wastewater (KW) was fairly toxic, while Sfax wastewater (SW) exhibited a high toxicity. This toxicity caused the inhibition of the anaerobic consortia when the MBR was fed with the SW. Moreover, the COD and the micro-toxicity increased during the day, parallel to the industrial and domestic activities resulting in the toxic character of SW during the day. SW treated in the AL remained toxic whereas the toxicity of KW treated in the AS or in anaerobic MBR decreased considerably. However, the anaerobic MBR showed a high efficiency in removing toxicity for both SW and KW. The quality of the anaerobic MBR permeate largely conforms with the microbiological WHO guidelines for unrestricted irrigation.

Disinfectant properties of essential oils from Salvia officinalis L. cultivated in Tunisia.

The essential oils were obtained by hydro-distillation of the aerial part of Salvia officinalis L. cultivated in Sfax gardens, Tunisia. The obtained oils were analyzed by gas chromatography-mass spectrometry (GC-MS) and 44 compounds were identified. Strong bactericidal and fungicidal effects were shown using the NCCLS broth dilution, EN 1275 and EN 1276 standard methods. The minimal cidal concentrations (MCCs) values ranged from 0.031 to 0.25 microLmL(-1). The essential oils concentrations of 0.5% and 1% (v/v) resulted in a reduction in viability higher than 5 and 4 log units per mL for the standard bacteria and fungi, respectively, within a contact time of 5 min. Using an air sampler and an aroma dispenser, vaporisation of 0.25 mLm(-3) of S. officinalis essential oils resulted in (72%, 73% and 70%) and (54%, 55% and 55%) reduction of the total microbial count and the total count of yeasts and moulds, after a residence time of 1h, 6h and 24h in a selected testing room, respectively. S. officinalis essential oils showed a potent vapour activity against a panel of bacteria, yeasts and fungi. This supported their use as a natural eco-friendly disinfectant to manage airborne microbes.

Assessment of toxicity of the untreated and treated olive mill wastewaters and soil irrigated by using microbiotests.

Hazard assessments based on two measures of toxicity were conducted for the untreated olive mill wastewaters (U), untreated olive mill wastewaters organic extract (UOE), treated olive mill wastewaters (T), treated olive mill wastewaters organic extract (TOE) and extracts of soils ferti-irrigated with untreated (SU) and with treated olive mill wastewaters (ST). The measures of toxicity were achieved by the determination of the bioluminescence inhibition percent (I(B)%) of Vibrio fischeri and by the growth inhibition (GI) of Bacillus megaterium, Pseudomonas fluorescens and Escherichia coli. A bioluminescence inhibition of V. fischeri of 100%, 100%, 65%, 47%, 46% and 30% were obtained with U, UOE, T, TOE, SU and ST respectively. Indeed, even diluted 24 times, a significant bioluminescence inhibition of 96% was obtained by U. However, only 30% bioluminescence inhibition was obtained by 24 times diluted T. Whereas, 24 times diluted, SU and ST did not show a bioluminescence inhibition (3% and 1%, respectively). The GI of B. megaterium, P. fluorescens and E. coli were, respectively, 93%, 72% and 100% by U; 100%, 80% and 100% by UOE; 70%, 60% and 89% by T; 63%, 54% and 68% by TOE; 39%, 27% and 43% by SU and 23%, 0% and 34% by ST. The incubation of U or T in the soil during four months reduced their toxicity by 54% and 35%, respectively. As it was expected, the most resistant bacterium to OMW toxicity is P. fluorescens then B. megaterium and E. coli. V. fischeri remained the most sensitive strain to the toxicity of this sewage what proves again its utilisation as standard of measure of the toxicity.

Polyphenols dynamics and phytotoxicity in a soil amended by olive mill wastewaters.

The effects of unprocessed olive mill wastewaters (OMW) on soil characteristics were investigated. Phenolic compounds levels in the treated soil were compared to those of a control soil profile. Results showed that OMW infiltration caused a modification of soil physicochemical characteristics. Phenolic compounds were detected at a depth of 1.2m four months after the last application of OMW. A moderate phytotoxic residual phenolic fraction (F) was extracted from the superficial soil layer 1 year after the OMW application. This residual F had a phytotoxic potential comparable to that of 25-fold diluted OMW.