Assessment of a UASB reactor with high ammonia concentrations: Effect of zeolite addition on process performance

The UASB process for wastewater treatment has been extensively studied, but the use of zeolite to improve UASB reactor performance has rarely been explored. In this study, a UASB reactor modified with natural zeolite operating at high nitrogen concentrations (0.5, 0.7 and 1 g/L) was evaluated. Two laboratory bioreactors, one with zeolite and one without, were operated at ambient temperatures ranging between 18 °C and 21 °C. The experimental phase had a start-up period of 21 days. In the reactor with zeolite, the pH was found to be between 7.9 and 9.1, with a COD removal efficiency of about 60% after 80 days of operation at ammonia concentrations of between 0.229 and 0.429 g/L in the effluent. In the reactor without zeolite, the pH was between 8.3 and 9.3, and the COD removal efficiency was about 40% at ammonia concentrations between 0.244 and 0.535 g/L in the effluent. The addition of zeolite also decreased the volatile suspended solids (VSS) concentration in the effluent, generating a biomass with larger granules and higher settling rates as compared to a UASB reactor without zeolite. Taking the lower ammonia concentration, the higher COD removal and the improved granulation into account, it can be concluded that natural zeolite positively influenced the behavior and performance of the UASB reactor operating with high nitrogen concentrations.     

A novel membrane-surface liquid co-culture to improve the production of laccase from Ganoderma lucidum

In this work, a laccase producer, Ganoderma lucidum, was separated and identified according to its morphological characteristics and phylogenetic data. A 4000 U/l and 8500 U/l of laccase activity was obtained in 500 ml flask by submerged culture and biomembrane-surface liquid culture (BSLC), respectively. Furthermore, the novel biomembrane-surface liquid co-culture (BSLCc) was developed by adding Saccharomyces cerevisiae to reactor in order to shorten the fermentation period and improve laccase production. Laccase activity obtained by BSLCc, 23 000 U/l, is 5.8 and 2.7 times of that obtained by submerged culture and BSLC, respectively. In addition, laccase production by BSLCc was successfully scaled-up to 100 l reactor, and 38 000 U/l of laccase activity was obtained on day 8. The mechanism of overproducing laccase by BSLCc was investigated by metabolism pathway analysis of glucose. The results show glucose limitation in fermentation broth induces the secretion of laccase. The addition of S. cerevisiae, on one hand, leads to an earlier occurrence of glucose limitation state, and thus shortens the fermentation time; on the other hand, it also results in the appearance of a series of metabolites of the yeast including organic acids, ethanol, glycerol and so forth in fermentation broth, and both polyacrylamide gel electrophoresis analysis and enzyme activity detection of laccase show that these metabolites contribute to the improvement of laccase activity.     

Improving biogas production from protein-rich distillery wastewater by decreasing ammonia inhibition

A large quantity of protein-rich distillery wastewater is produced during the process of bio-ethanol production from kitchen waste. It is difficult, however, to treat protein-rich distillery wastewater by anaerobic digestion due to ammonia inhibition. In this study, a novel method was investigated to reduce ammonia inhibition during thermophilic anaerobic digestion through the recirculation of water-washed biogas into the headspace (R1 system) or liquid phase (R2 system) of the reactors. The results show that the method greatly improved biogas production from distillery wastewater. R2 system achieved stable biogas production at a higher organic loading rate (OLR) of 4.0 g VTS/L/d than R1 system at 3.0 g VTS/L/d. At the same OLR, we observed a higher biogas production rate but lower accumulation of NH₄⁺ and volatile fatty acids in the reactor, and higher ammonia absorption rate in the water tank of R2 system than R1 system. The better performance of R2 system could be attributed to the more efficient removal of ammonia from liquid phase. In addition, adjusting the C/N ratio of distillery wastewater from 9.0 to 11.4 significantly enhanced the maximum OLR from 3.0 to 7.0 g VTS/L/d in R1 system.     

Involvement and interaction of microbial communities in the transformation and stabilization of chromium during the composting of tannery effluent treated biomass of Vallisneria spiralis L.

Tannery effluent treated with aquatic macrophyte Vallisneria spiralis L. for 14 d showed significant improvement in physico-chemical properties and reduction in Cr concentration. Accumulation of Cr was found maximum in roots (358 μg g^?1dw) as compared to shoot (62 μg g^?1dw) of the plant. A laboratory scale composter was designed with the objectives to investigate the physico-chemical changes and role of microbes in stabilization and transformation of Cr in the composting material. Results revealed that the composting process was quick within 7–21 d as indicated by peak time for various physico-chemical parameters and drop in C/N ratio up to acceptable limit. The profile of microbial communities indicated that population of anaerobic, aerobic and nitrifying bacteria increased quickly at the initial phase, and reached a peak level of 4.2 × 10⁶, 9.78 × 10⁸ and 9.32 × 10⁹ CFU g^?1, respectively at 21 d; while population of actinomycetes and fungi was found maximum i.e. 3.29 × 10⁷ and 9.7 × 10⁶ CFU g^?1, respectively, after 35 d of composting. Overall bacterial population dominated over the actinomycetes and fungi during the composting process. Cr^(VI) was transformed to Cr^(III) due to the microbial activity during the process. Sequential extraction of Cr fractionation showed its stabilization via changing into organic matter-bound and residual fractions during the composting.     

Nitrogen amendment of green waste impacts microbial community,enzyme secretion and potential for lignocellulose decomposition

Microorganisms involved in biomass deconstruction are an important resource for organic waste recycling and enzymes for lignocellulose bioconversion. The goals of this study were to examine the impact of nitrogen amendment on microbial community restructuring, secretion of xylanases and endoglucanases, and potential for biomass deconstruction. Communities were cultivated aerobically at 55 °C on green waste (GW) amended with varying levels of NH₄Cl. Bacterial and fungal communities were determined using 16S rRNA and ITS region gene sequencing and PICRUSt (Phylogenetic Investigation of Communities by Reconstruction of Unobserved States) was applied to predict relative abundance of genes involved in lignocellulose hydrolysis. Nitrogen amendment significantly increased secretion of xylanases and endoglucanases, and microbial activity; enzyme activities and cumulative respiration were greatest when nitrogen level in GW was between 4.13–4.56 wt% (g/g), but decreased with higher nitrogen levels. The microbial community shifted to one with increasing potential to decompose complex polymers as nitrogen increased with peak potential occurring between 3.79–4.45 wt% (g/g) nitrogen amendment. The results will aid in informing the management of nitrogen level to foster microbial communities capable of secreting enzymes that hydrolyze recalcitrant polymers in lignocellulose and yield rapid decomposition of green waste.     

A half-submerged integrated two-phase anaerobic reactor for agricultural solid waste codigestion

Anaerobic digestion is widely used in bioenergy recovery from waste. In this study, a half-submerged, integrated, two-phase anaerobic reactor consisting of a top roller acting as an acidogenic unit and a recycling bottom reactor acting as a methanogenic unit was developed for the codigestion of wheat straw (WS) and fruit/vegetable waste (FVW). The reactor was operated for 21 batches (nearly 300 d). Anaerobic granular sludge was inoculated into the methanogenic unit. The residence time for the mixed waste was maintained as 10 d when the operation stabilized, and the temperature was kept at 35 °C. The highest organic loading rate was 1.37 kg VS/(m³ d), and the maximum daily biogas production was 328 L/d. Volatile solid removal efficiencies exceeded 85%. WS digestion could be confirmed, and efficiency was affected by both the ratio of WS to FVW and the loading rate. The dominant bacteria were Bacteroides-like species, which are involved in glycan and cellulose decomposition. Methanogenic community structures, pH levels, and volatile fatty acid concentrations in the acidogenic and methanogenic units differed, indicating successful phase separation. This novel reactor can improve the mass transfer and microbial cooperation between acidogenic and methanogenic units and can efficiently and steady codigest solid waste.     

NOB suppression and adaptation strategies in the partial nitrification–Anammox process for a poultry manure anaerobic digester

Poultry manure contains high levels of ammonia, which result in a suboptimal bioconversion to methane in anaerobic digesters (AD). A simultaneous process of nitrification, Anammox and denitrification (SNAD) in a continuous granular bubble column reactor to treat the anaerobically digested poultry manure was implemented. Thus, two strategies to achieve high efficiencies were proposed in this study: (1) ammonia overload to suppress nitrite oxidizing bacteria (NOB) and (2) gradual adaptation of the partial nitrification–Anammox (PN–A) biomass to organic matter. During the NOB-suppression stage, microbial and physical biomass characterizations were performed and the NOB abundance decreased from 31.3% to 3.3%. During the adaptation stage, with a nitrogen loading rate of 0.34 g L⁻¹ d⁻¹, a hydraulic retention time of 1.24 d and an influent COD/N ratio of 2.63 ± 0.02, a maximum ammonia and total nitrogen removal of 100% and 91.68% were achieved, respectively. The relative abundances of the aerobic and the anaerobic ammonia-oxidizing bacteria were greater than 35% and 40% respectively, during the study. These strategies provided useful design tools for the efficient removal of nitrogen species in the presence of organic matter.     

Influence on dithiolopyrrolone antibiotic production by organic acids in Saccharothrix algeriensis NRRL B-24137

The influence of organic acids on growth and dithiolopyrrolone antibiotic production by Saccharothrix algeriensis NRRL B-24137 was studied. The production of dithiolopyrrolones depends upon the nature and concentration of the organic acids in the culture medium. Study of the nature of organic acids showed that the most effective organic acids for thiolutin specific production were maleic, 4-hydroxybenzoic, benzentetracarboxylic, pantothenic, pivalic and pyruvic acids (which yielded almost five-fold over the starting medium) and pimelic acid (more than three-fold). 4-Bromobenzoic acid showed the best production of senecioyl-pyrrothine (59 mg g⁻¹ DCW). Tiglic acid showed the best production of tigloyl-pyrrothine (22 mg g⁻¹ DCW). The highest yield of isobutyryl-pyrrothine (7.6 mg g⁻¹ DCW) was observed in the presence of crotonic acid. Sorbic acid yielded the best production of butanoyl-pyrrothine (26 mg g⁻¹ DCW). Methacrylic, butyric, pyruvic and 4-bromobenzoic acids also exhibited the best production of butanoyl-pyrrothine (27–11-fold).Study of organic acid concentration showed that among the selected organic acids, pimelic acid yielded the highest specific production of thiolutin (91 mg g⁻¹ DCW) at 7.5 mM; and senecioyl-pyrrothine (11 mg g⁻¹ DCW), tigloyl-pyrrothine (9 mg g⁻¹ DCW) and butanoyl-pyrrothine (3.5 mg g⁻¹ DCW) at 5 mM. Pyruvic acid at 1.25 mM enhanced the production of senecioyl-pyrrothine (4.3 mg g⁻¹ DCW). The maximum production of tigloyl-pyrrothine (18.6 mg g⁻¹ DCW) was observed in the presence of tiglic acid at 2.5 mM. Maximum production of isobutyryl-pyrrothine was observed in the presence of 7.5 mM tiglic acid. In addition, methacrylic acid (at 5 mM) and butyric acid (at 2.5 mM) enhanced the production of butanoyl-pyrrothine (26 and 20 times, respectively).The above results can be employed in the optimisation of the culture medium for the production of dithiolopyrrolone in higher quantities.     

Decomposition processes in soil of a healthy and a declining Phragmites australis stand

Decomposition processes were investigated in the soil of a declining, more eutrophic and a healthy, less eutrophic freshwater reed (Phragmites australis (Cav.) Trin. ex Steudel) stand in the littoral zone of Rožmberk fishpond, Czech Republic. Soil and pore water were sampled five times from April to October 1998. Chemical properties, CO₂ production in oxic and anoxic conditions, CH₄ production, denitrifying enzyme activity (DEA) and bacterial biomass were measured under laboratory conditions in suspensions prepared from homogenised soil samples. The more eutrophic West stand was more anaerobic than the East stand, with lower redox potential, lower pH and with a higher amount of organic acids, mainly acetic and lactic acid. Mean seasonal concentrations of total nitrogen in pore water, nitrogen of amino acids and proteins, and reducing sugars were all higher in the soil at the more eutrophic stand. Higher nutrient status and more reduced conditions at the more eutrophic stand were accompanied by (i) a limitation of aerobic microbial activities (CO₂ production in oxic conditions: 0.35 versus 0.54 μmol CO₂ cm⁻³ h⁻¹); lower DEA (4.0 versus 20.2 nmol N₂O cm⁻³ h⁻¹) and a lower proportion of bacteria that were active in aerobic conditions; (ii) by a prevalence of anaerobic over aerobic microbial processes; (iii) by a higher rate of methanogenesis (15.0 versus 11.5 nmol CH₄ cm⁻³ h⁻¹) and (iv) by an overall lower rate of microbial processes as compared to less eutrophied stand. The shift from aerobic to anaerobic microbial metabolism, and a coinciding restriction of metabolic activities at the more eutrophic stand are indicative of an elevated oxygen stress in the soil, associated with accumulation of metabolites toxic to both the micro-organisms and the reed. Possible links between eutrophication, decomposition processes in the soil and reed decline are discussed.     

Improvement in natamycin production by Streptomyces natalensis with the addition of short-chain carboxylic acids

Natamycin is an important tetraene (polyene) antibiotic produced in submerged culture by different strains of Streptomyces sp. In the present work, the effects of the addition of short-chain carboxylic acids (acetic, propionic and butyric) on cell growth and the kinetics of natamycin production were investigated during submerged cultivation of Streptomyces natalensis. The addition of acetic and propionic acids showed stimulatory effects on natamycin production when added to the fermentation medium at concentrations below 2 g L^?1 at the beginning of cultivation. In addition, when acetic and propionic acids were added in a mixture (7:1) at a total concentration of 2 g L^?1, antibiotic production increased significantly, reaching 3.0 g L^?1 (approximately 223% and 250% increases in volumetric and specific antibiotic production, respectively, compared with the control culture). Moreover, the addition of carboxylic acids not only increased the antibiotic yield but also decreased the production time from 96 h to only 84 h in shake-flask cultures. A further enhancement in natamycin production was achieved by cultivation in a 2-L stirred-tank bioreactor under controlled pH conditions. The maximum volumetric production of 3.98 g L^?1 was achieved after 84 h in carboxylic acid-supplemented culture (acetate and propionate in a ratio of 7:1).     

Operation of a fixed-bed bioreactor in batch and fed-batch modes for production of inulinase by solid-state fermentation

This work is focused on the inulinase production by solid-state fermentation (SSF) in a fixed-bed reactor (34 cm diameter and 50 cm height) with working capacity of 2-kg of dry substrate operated in batch and fed-batch modes. It was investigated different strategies for feeding the inlet air in the bioreactor (saturated and unsaturated air) as alternative to remove the metabolic heat generated during the microbial growth by evaporative cooling. The kinetic evaluation of the process carried out in batch mode using unsaturated air showed that the evaporative cooling decreasing the mean temperature of the solid-bed, although the enzyme production was lower than that obtained using saturated air. Results showed that maximum enzyme activity (586 ± 63 U gds⁻¹) was obtained in the fed-batch mode using saturated air after 24 h of fermentation. The enzymatic extract obtained by fed-batch mode was characterized and presented optimum temperature and pH in the range of 52–57 °C and 4.8–5.2, respectively. For a temperature range from 40 to 70 °C the enzyme presented decimal reduction time, D-value, ranging from 5748 to 47 h, respectively. For a pH range from 3.5 to 5.5 the enzyme showed good stability, presenting D-values higher than 2622 h. In terms of Michaelis–Mentem parameters were demonstrated that the crude inulinase activity presented higher affinity for substrate sucrose compared to inulin.     

Operation of a bioelectrochemical system as a polishing stage for the effluent from a two-stage biohydrogen and biomethane production process

Anaerobic bioenergy production processes including fermentative biohydrogen (BioH₂), anaerobic digestion (AD) and bioelectrochemical system have been investigated for converting municipal waste or various biomass feedstock to useful energy carriers. However, the performance of a microbial fuel cell (MFC) fed on the effluent from a two-stage biogas production process has not yet been investigated extensively in continuous reactor operation on complex substrates. In this study we have investigated the extent to which a microbial fuel cell (MFC) can reduce COD and recover further energy from the effluent of a two-stage biohydrogen and biomethane system. The performance of a four-module tubular MFC was determined at six different organic loadings (0.036–6.149 g sCOD L⁻¹ d⁻¹) in terms of power generation, COD removal efficiency, coulombic efficiency (CE) and energy conversion efficiency (ECE). A power density of 3.1 W m⁻³ was observed at the OLR = 0.572 g sCOD L⁻¹ d⁻¹, which resulted in the highest CE (60%) and ECE (0.8%), but the COD removal efficiency decreased at higher organic loading rates (35.1–4.4%). The energy recovery was 92.95 J L⁻¹ and the energy conversion efficiency, based on total influent COD was found to be 0.48–0.81% at 0.572 g sCOD L⁻¹ d⁻¹. However, the energy recovery by the MFC is only reported for a four-module reactor and improved performance can be expected with an extended module count, as chemical energy remained available for further electrogenesis.     

Influence of condensed tannins from Brazilian semi-arid legumes on ruminal degradability,microbial colonization and ruminal enzymatic activity in Saanen goats

The present study aimed at determining the influence of condensed tannins present in the Brazilian legume species Mimosa hostilis, Mimosa caesalpinifolia and Bauhinia cheilantha on ruminal degradability, microbial colonization and enzymatic activity. Polyethylene glycol (PEG) was used to reduce the astringency and concentration of soluble condensed tannins. Four ruminally-cannulated Saanen goats (60 ± 8 kg BW) were fed, in two experimental periods, with a hay diet based on the studied legumes treated or non-treated with PEG. Voluntary intake, microbial colonization, DM, CP, NDF, and ruminal degradability of PEG treated and non-treated forage leaves, as well as pH, ammonia and 1,4 β-endoglucanase activity of the rumen content were evaluated. Astringency and soluble tannin concentration of the studied legumes were reduced by approximately 70% and 50%, respectively, with PEG treatment. Average DM intake was higher for the treated diet (16.76 g DM/kg BW/day against 13.06 g DM/kg BW/day). Percentile values for degradation parameters and for potential and effective degradabilities of DM, CP and NDF were also affected by the tannins, but at different intensities. Electron microscopic observations of ruminally-incubated legume leaves showed a more effective microbial colonization of PEG-treated leaves for all legume species. A decrease in pH and an increase in ammonia concentration and in endoglucanase activity in the ruminal content was also observed for PEG-treated diets at all sampling periods. Condensed tannins of the studied legume species have influenced the adhesion conditions, colonization and enzymatic activity of the microbial ecosystem, and consequently the ruminal degradation of the different dietary fractions. For this reason, the reduction in condensed tannin would be of great importance to improve the nutrition of ruminant feeding of these species.     

Bioavailability and extraction of heavy metals from contaminated soil by Atriplex halimus

Pot experiments were performed to evaluate the phytoremediation capacity of plants of Atriplex halimus grown in contaminated mine soils and to investigate the effects of organic amendments on the metal bioavailability and uptake of these metals by plants. Soil samples collected from abandoned mine sites north of Madrid (Spain) were mixed with 0, 30 and 60 Mg ha⁻¹ of two organic amendments, with different pH and nutrients content: pine-bark compost and horse- and sheep-manure compost. The increasing soil organic matter content and pH by the application of manure amendment reduced metal bioavailability in soil stabilising them. The proportion of Cu in the most bioavailable fractions (sum of the water-soluble, exchangeable, acid-soluble and Fe–Mn oxides fractions) decreased with the addition of 60 Mg ha⁻¹ of manure from 62% to 52% in one of the soils studied and from 50% to 30% in the other. This amendment also reduced Zn proportion in water-soluble and exchangeable fractions from 17% to 13% in one of the soils. Manure decreased metal concentrations in shoots of A. halimus, from 97 to 35 mg kg⁻¹ of Cu, from 211 to 98 mg kg⁻¹ of Zn and from 1.4 to 0.6 mg kg⁻¹ of Cd. In these treatments there was a higher plant growth due to the lower metal toxicity and the improvement of nutrients content in soil. This higher growth resulted in a higher total metal accumulation in plant biomass and therefore in a greater amount of metals removed from soil, so manure could be useful for phytoextraction purposes. This amendment increased metal accumulation in shoots from 37 to 138 mg pot⁻¹ of Cu, from 299 to 445 mg pot⁻¹ of Zn and from 1.8 to 3.7 mg pot⁻¹ of Cd. Pine bark amendment did not significantly alter metal availability and its uptake by plants. Plants of A. halimus managed to reduce total Zn concentration in one of the soils from 146 to 130 mg kg⁻¹, but its phytoextraction capacity was insufficient to remediate contaminated soils in the short-to-medium term. However, A. halimus could be, in combination with manure amendment, appropriate for the phytostabilization of metals in mine soils.     

Varying endophyte status and energy supplementation of fresh tall fescue in continuous culture

Eight dual-flow continuous culture vessels (700 ml) were used to compare in vitro effects of toxic, endophyte-infected (E+), endophyte-free (E−), and non-toxic, endophyte-infected (EN) Jesup tall fescue (vegetative stage) on ruminal fermentation at 4 levels (0, 150, 300, and 450 g kg⁻¹ DM) of concentrate supplementation (ground corn) for a total of 12 experimental diets in a randomized incomplete block design with 2 replicates. Each culture vessel was offered a total of 15 g DM d⁻¹. Forage was fed in four equal portions (fed at 03:00, 09:00, 15:00, and 21:00 h); and corn was fed in two equal portions (fed at 09:00 and 21:00 h). Headspace gas and liquid samples were analyzed for methane, ruminal culture pH, ammonia–N, and volatile fatty acid production. Ammonia–N output (g d⁻¹) varied by grass; EN had lower values compared to those of E+ and E−. Increasing the level of grain linearly decreased ruminal culture pH, ammonia–N, acetate production, and the acetate-to-propionate ratio, whereas propionate and butyrate production increased with higher grain supplementation. Ruminal fermentation was minimally altered by the presence of the endophyte; however, for the highest level of grain fed (450 g kg⁻¹ DM fed) the methane production pattern for all three grasses was altered. In addition to having the lowest ruminal ammonia–N accumulation, the non-toxic, endophyte-infected fescue resulted in the lowest methane production measured.     

Characteristics of nitrifying granules developed in an air pulsing SBR

Nitrifying bacteria are characterized by low biomass yields and slow activity rates, the enhancement of nitrifying biomass retention by promoting the formation of granules would allow improving nitrification process. The use of a pulsing sequencing batch reactor (SBR) was tested to produce nitrifying granules minimizing the air consumption. After 400 days of operation, 1.07 g VSS/L of granular biomass with a mean diameter of 0.9 mm was accumulated in the system. The maximal nitrate production rate reached a value of 0.3 kg N/(m³ d).Sporadic nitrite accumulations were registered and attributed to oxygen diffusion limitations which were solved improving the granule-liquid mass transfer with a liquid recirculation system. FISH technique was used in order to identify the main populations in the nitrifying granules; Nitrosomonas spp. and Nitrospira spp. were identified as the main ammonia and nitrite oxidizing population, respectively. With the introduction of liquid recirculation in the reactor the ammonia oxidation efficiencies improved and the proportion of Nitrospira spp. increased from 1% to 15%.     

Simultaneous decrease in ammonia and hydrogen sulfide inhibition during the thermophilic anaerobic digestion of protein-rich stillage by biogas recirculation and air supply at 60 °C

We had proposed a novel method to reduce ammonia inhibition during thermophilic anaerobic digestion via recirculation of water-washed biogas into the headspace (R1 system) or liquid phase (R2 system) of reactors. The feasibility of reducing the ratio of recirculated biogas to biogas produced (called the biogas recirculation ratio) was investigated in the present study. Thermophilic anaerobic digestion at 53 °C and 60 °C with a biogas recirculation ratio of 150 facilitated stable digestion performance and biogas production at a higher organic loading rate of 7 g/L/d in the R1 system, while the ammonia removal efficiency increased 1.23-fold when the temperature increased from 53 °C to 60 °C. At 60 °C, the biogas recirculation ratios in the R1 and R2 systems decreased to 50 and 10, and the ammonia absorption rates were 6.1 and 8.3 mmol/L/d, respectively, without decreasing the anaerobic digestion performance. The ammonia absorption rate of 8.3 mmol/L/d in the R2 system was higher than the rate of 7.8 mmol/L/d at the biogas recirculation ratio of 150 in the R1 system. The hydrogen sulfide content in the biogas was reduced to less than 50 ppm by supplying air at 3% of the amount of biogas produced into the reactor.     

Highly soluble expression and molecular characterization of an organic solvent-stable and thermotolerant lipase originating from the metagenome

A novel organic solvent-stable and thermotolerant lipase gene (designated ostl28) was cloned from a metagenomic library and overexpressed in Escherichia coli BL21 (DE3) in soluble form. OSTL28 contained 262 amino acids with relative molecular mass 30.1 kDa and isoelectric point 9.7. The optimum pH and temperature of the OSTL28 were 7.5 and 60 °C, respectively. OSTL28 was stable in the pH range of 4.5–9.5 and at temperatures below 65 °C. The enzyme could hydrolyze a wide range of ρ-nitrophenyl esters, but its best substrate is ρ-nitrophenyl laurate with the highest activity of 236 U/mg (54,000 U/L). The recombinant OSTL28 was highly resisted to organic solvents, especially glycerol and methanol. The metal ions, with the exception of Hg²⁺ and Ag⁺, did not have any influence on enzyme activity, whereas non-ionic surfactants and Al³⁺ slightly activated the enzyme. These features indicate that it is a potential biocatalyst for biodiesel production.     

Operation of an aerobic granular pilot scale SBR plant to treat swine slurry

A pilot-scale Sequencing Batch Reactor was operated during 307 days in order to treat swine slurry characterized by its high variable composition: organic and nitrogen applied loading rates and C/N ratio were 1.4–6.3 kg COD_s/(m³ d), 0.5–2.5 kg N/(m³ d) and 1.9–9.4 g COD_s/(g N), respectively. Aerobic granules successfully developed in the reactor and their physical properties remained rather stable despite the feeding composition variability. Organic and ammonia removal efficiency reached 61–73% and 56–77%, respectively, however ammonia was mainly oxidized to nitrite. The reactor had a good biomass retention capacity to select for granular biomass. However, its efficiency to retain the solids present in the feeding was low. Aerobic granulation in SBR systems appears as an interesting alternative to treat slurry in small livestock facilities where the implementation of anaerobic digestion systems is not a feasible option or the removal of nitrogenous compounds is required.     

Real-time control strategy for nitrogen removal via nitrite in a SHARON reactor using pH and ORP sensors

This paper presents a real-time control strategy for nitrogen removal via nitrite in a continuous flow SHARON reactor using on-line available and industrially feasible sensors (pH and ORP). The developed control strategy optimizes the length of aerobic and anoxic phases as well as the external carbon source addition. This strategy, implemented in a laboratory-scale SHARON reactor fed with synthetic wastewater and real dewatering sludge supernatant, was able to cope with step variations in influent flow rate and ammonium concentration. The main advantages of this control strategy over the traditional operation mode with fixed carbon source dosification and fixed length cycle operation were: better effluent quality (ammonia concentration decreased from 12 to 2 mg NH₄–N L^?1 and nitrogen removal efficiency raised from 95% to 98%) as result of the shorter cycle length: 2.9 h versus 4.0 h, and savings in external carbon addition: 1332 mg COD d^?1 versus 2100 mg COD d^?1.