Development of aerobic granular sludge under tropical climate conditions: The key role of inoculum adaptation under reduced sludge washout for stable granulation.

Aerobic granular sludge (AGS) is a promising technology for wastewater treatment. However, the success of the process depends on the formation of stable granular biomass, which is associated with the microbiological aspects of the sludge and reactor operating conditions. In this study, the development of AGS from a poor nitrifying flocculent sludge obtained in a sewage treatment plant designed only for organic matter removal was assessed in a sequencing batch reactor (SBR) under tropical climate conditions (temperatures of 28 ±â€¯4 °C). The results showed that, despite the alternating anaerobic-aerobic conditions during the granules selection phase under high sludge washout rates (low settling time), readily biodegradable organic matter was mainly removed aerobically. The formed granules were unstable, exhibiting a substantial amount of filaments and pasty consistency. The biomass characteristics (e.g., sludge volume index, density, diameter and settling velocity) were negatively impacted as complete granulation was reached, while biomass loss and degranulation became inevitable. Poor nitrification and no enhanced biological phosphate removal (EBPR) were observed. Implementation of a new operational strategy incorporating an adaptation of the seed sludge under reduced washout conditions (high settling time) prior to the granules selection stage enabled most of the influent organics to be removed anaerobically. Besides allowing a feast-famine regime to be established in the reactor, the sludge acclimation phase favoured the development of slow-growing organisms and suppressed the appearance of filamentous-like structures. Fast-settling granules with regular shape remained stable in the long-term, while high ammonium (>95%) and total nitrogen removal (>90%) was obtained. However, EBPR activity was very unstable, most likely due to the high temperatures. The findings of this study are important for the spreading of the AGS technology worldwide, especially in developing countries where the conditions are different in all aspects.

Interpreting the effect of increasing COD loading rates on the performance of a pre-anoxic MBBR system: implications on the attached and suspended biomass dynamics and nitrification-denitrification activity.

A pre-anoxic MBBR system was subjected to increasing organic loading rates up to 18 gCOD/(m(2) day). At 3 gCOD/(m(2) day), most of the incoming organic matter was removed via denitrification. However, at higher loads, anoxic COD removal became limited by the nitrite/nitrate supply from the aerobic reactor, which assumed an important role in this conversion. Despite the application of low dissolved oxygen (DO) levels (<2 mg/L) in this tank, nitrification was observed to be nearly complete until 8 gCOD/(m(2) day). As the organic input was increased, the maximum specific nitrifying activity gradually declined. Activity tests suggested that an oxygen-limited environment was established in the biofilm. At lower loads [3-8 gCOD/(m(2) day)], the nitrification product obtained was affected by the DO concentration, whereas from 16 to 21 gCOD/(m(2) day), nitrite/nitrate profiles were likely associated with microbial stratification in the biofilm. The results also indicated that the role of the suspended biomass in the overall nitrification and denitrification can be very significant in high loaded MBBRs and should not be neglected, even at low HRTs.

Integration of biofiltration and advanced oxidation processes for tertiary treatment of an oil refinery wastewater aiming at water reuse.

The combination of biological and chemical oxidation processes is an interesting approach to remove ready, poor, and non-biodegradable compounds from complex industrial wastewaters. In this study, biofiltration followed by H2O2/UV oxidation (or microfiltration) and final reverse osmosis (RO) step was employed for tertiary treatment of an oil refinery wastewater. Biofiltration alone allowed obtaining total organic carbon (TOC), chemical oxygen demand (COD), UV absorbance at 254 nm (UV254), ammonium, and turbidity removal of around 46, 46, 23, 50, and 61 %, respectively. After the combined biological-chemical oxidation treatment, TOC and UV254 removal amounted to 88 and 79 %, respectively. Whereas, the treatment performance achieved with different UV lamp powers (55 and 95 W) and therefore distinct irradiance levels (26.8 and 46.3 mW/cm(2), respectively) were very similar and TOC and UV254 removal rates were highly affected by the applied C/H2O2 ratio. Silt density index (SDI) was effectively reduced by H2O2/UV oxidation, favoring further RO application. C/H2O2 ratio of 1:4, 55 W UV lamp, and 20-min oxidation reaction corresponded to the experimental condition which provided the best cost/benefit ratio for TOC, UV254, and SDI reduction from the biofilter effluent. The array of treatment processes proposed in this study has shown to be adequate for tertiary treatment of the oil refinery wastewater, ensuring the mitigation of membrane fouling problems and producing a final effluent which is suitable for reuse applications.

A comparison between the oxidation with laccase and horseradish peroxidase for triclosan conversion.

Triclosan is a broad-spectrum biocide used in personal-care products that is suspected to be linked to the emergence of antibiotic-resistant bacteria. In the present work, the enzymes horseradish peroxidase and laccase from Trametes versicolor were evaluated for the conversion of triclosan in an aqueous matrix. The removal of antibacterial activity by the enzymatic processes was evaluated by an assay based on the growth inhibition of Escherichia coli K12. The horseradish peroxidase (HRP) process appears more advantageous than the laccase process in removing triclosan from an aqueous matrix, considering the reaction parameters pH, temperature, catalytic efficiency, and enzyme concentration. The highest conversion of triclosan catalysed by laccase was observed at pH 5.0, that is, lower than the typical pH range (6.5-7.5) of sewage treatment plants' effluents. The efficiency of laccase process was much more impacted by variations in the temperature in the range of 10-40°C. Kinetic studies showed that triclosan is a substrate more specific for HRP than for laccase. The protein content for the HRP-catalysed process was 14 times lower than that for the laccase process. Decay kinetics suggest that reaction mechanisms depend on enzyme concentration and its concentration. Both processes were able to reduce the antibacterial activity, and the residual activity of the treated solution is probably due to non-converted triclosan and not due to the reaction products. The laccase-catalysed conversion of triclosan in an environmental relevant concentration required a higher amount of enzyme than that required in the HRP process.

Combined organic matter and nitrogen removal from a chemical industry wastewater in a two-stage MBBR system.

Pesticide-producing factories generate highly polluting wastewaters containing toxic and hazardous compounds which should be reduced to acceptable levels before discharge. In this study, a chemical industry wastewater was treated in a pre-denitrification moving-bed biofilm reactor system subjected to an increasing internal mixed liquor recycle ratio from 2 to 4. Although the influent wastewater characteristics substantially varied over time, the removal of chemical oxygen demand (COD) and dissolved organic carbon was quite stable and mostly higher than 90%. The highest fraction of the incoming organic matter was removed anoxically, favouring a low COD/N environment in the subsequent aerobic nitrifying tank and thus ensuring stable ammonium removal (90-95%). However, during pH and salt shock periods, nitrifiers were severely inhibited but gradually restored their full nitrifying capability as non-stressing conditions were reestablished. Besides promoting an increase in the maximum nitrification potential of the aerobic attached biomass from 0.34 to 0.63 mg [Formula: see text], the increase in the internal recycle ratio was accompanied by an increase in nitrogen removal (60-78%) and maximum specific denitrification rate (2.7-3.3 mg NOx(-)--N). Total polysaccharides (PS) and protein (PT) concentrations of attached biomass were observed to be directly influenced by the influent organic loading rate, while the PS/PT ratio mainly ranged from 0.3 to 0.5. Results of Microtox tests showed that no toxicity was found in the effluent of both the anoxic and aerobic reactors, indicating that the biological process was effective in removing residual substances which might adversely affect the receiving waters' ecosystem.

Tracking the dynamics of heterotrophs and nitrifiers in moving-bed biofilm reactors operated at different COD/N ratios.

In this study, the impact of COD/N ratio and feeding regime on the dynamics of heterotrophs and nitrifiers in moving-bed biofilm reactors was addressed. Based on DGGE analysis of 16S rRNA genes, the influent COD was found to be the main factor determining the overall bacterial diversity. The amoA-gene-based analysis suggested that the dynamic behavior of the substrate in continuous and pulse-feeding reactors influenced the selection of specific ammonium-oxidizing bacteria (AOB) strains. Furthermore, AOB diversity was directly related to the applied COD/N ratio and ammonium-nitrogen load. Maximum specific ammonium oxidation rates observed under non-substrate-limiting conditions were observed to be proportional to the fraction of nitrifiers within the bacterial community. FISH analysis revealed that Nitrosomonas genus dominated the AOB community in all reactors. Moreover, Nitrospira was found to be the only nitrite-oxidizing bacteria (NOB) in the fully autotrophic system, whereas Nitrobacter represented the dominant NOB genus in the organic carbon-fed reactors.

Treatment of petroleum refinery wastewater containing heavily polluting substances in an aerobic submerged fixed-bed reactor.

Petroleum refineries produce large amount of wastewaters, which often contain a wide range of different compounds. Some of these constituents may be recalcitrant and therefore difficult to be treated biologically. This study evaluated the capability of an aerobic submerged fixed-bed reactor (ASFBR) containing a corrugated PVC support material for biofilm attachment to treat a complex and high-strength organic wastewater coming from a petroleum refinery. The reactor operation was divided into five experimental runs which lasted more than 250 days. During the reactor operation, the applied volumetric organic load was varied within the range of 0.5-2.4 kgCOD.m(-3).d(-1). Despite the inherent fluctuations on the characteristics of the complex wastewater and the slight decrease in the reactor performance when the influent organic load was increased, the ASFBR showed good stability and allowed to reach chemical oxygen demand, dissolved organic carbon and total suspended solids removals up to 91%, 90% and 92%, respectively. Appreciable ammonium removal was obtained (around 90%). Some challenging aspects of reactor operation such as biofilm quantification and important biofilm constituents (e.g. polysaccharides (PS) and proteins (PT)) were also addressed in this work. Average PS/volatile attached solids (VAS) and PT/VAS ratios were around 6% and 50%, respectively. The support material promoted biofilm attachment without appreciable loss of solids and allowed long-term operation without clogging. Microscopic observations of the microbial community revealed great diversity of higher organisms, such as protozoa and rotifers, suggesting that toxic compounds found in the wastewater were possibly removed in the biofilm.

Oestrogenicity assessment of s-triazines by-products during ozonation.

The triazines are a group of herbicides with a wide range of uses. Atrazine is, in fact, one of the most used agricultural pesticides in the world. The terbuthylazine is applied as a substitute of atrazine in some countries of Europe since 2004, when the European Union announced a ban of atrazine because of ubiquitous water contamination. In this study, both atrazine and terbuthylazine were degraded by the ozone process to estimate the efficiency on pesticide removal in water, the intermediates formed and their potential oestrogenic activity using the yeast oestrogen screen (YES) test. Both pesticides were rapidly eliminated from the medium during ozonation (applied ozone dose 0.083 and 0.02 mmol O3 L(-1), respectively). The results show that both compounds generated similar by-products from ozone degradation. Moreover, significant oestrogenic activity was detected for both atrazine and terbuthylazine intermediates, during the first minutes of ozonation. The YES assay used in this study proved to be a sensitive tool in assessing trace amounts of oestrogenic chemicals, which can represent critical issues influencing the experimental results in environmental applications.

Measuring biomass specific ammonium, nitrite and phosphate uptake rates in aerobic granular sludge.

Aerobic granular sludge (AGS) technology offers the possibility to remove organic carbon, nitrogen and phosphorus in a single reactor system. The granular structure is stratified in such a way that both aerobic and anaerobic/anoxic layers are present. Since most of the biological processes in AGS systems occur simultaneously, the measurement and estimation of the capacity of specific conversions is complicated compared to suspended biomass. The determination of the activities of different functional groups in aerobic granular sludge allows for identification of the potential metabolic capacity of the sludge and aids to analyze bioreactor performance. It allows for comparison of different sludges and enables improved understanding of the interaction and competition between different metabolic groups of microorganisms. The most appropriate experimental conditions and methods to determine specific ammonium, nitrite and phosphate uptake rates under normal operation of AGS reactors were evaluated and described in this study. Extra biomass characterization experiments determining the maximum uptake rate of these compounds on optimized conditions were performed as well to see how much spare capacity was available. The methodologies proposed may serve as an experimental frame of reference for investigating the metabolic capacities of microbial functional groups in biofilm processes.

Simultaneous nitrogen and phosphate removal in aerobic granular sludge reactors operated at different temperatures.

The main biological conversions taking place in two lab-scale aerobic granular sludge sequencing batch reactors were evaluated. Reactors were operated at different temperatures (20 and 30 °C) and accomplished simultaneous COD, nitrogen and phosphate removal. Nitrogen and phosphate conversions were linked to the microbial community structure as assessed by fluorescent in situ hybridization (FISH) analysis. Anoxic tests were performed to evaluate the contribution of anoxic phosphate uptake to the overall phosphate removal and to clarify the denitrification pathway. Complete nitrification/denitrification and phosphate removal were achieved in both systems. A considerable fraction of the phosphate removal was coupled to denitrification (denitrifying dephosphatation). From the results obtained in anoxic batch experiments dosing either nitrite or nitrate, denitrification was proposed to proceed mainly via the nitrate pathway. Denitrifying glycogen-accumulating organisms (DGAOs) were observed to be the main organisms responsible for the reduction of nitrate to nitrite. A significant fraction of the nitrite was further reduced to nitrogen gas while being used as electron acceptor by denitrifying polyphosphate-accumulating organisms (PAO clade II) for anoxic phosphate uptake.

Improved phosphate removal by selective sludge discharge in aerobic granular sludge reactors.

Two lab-scale aerobic granular sludge sequencing batch reactors were operated at 20 and 30°C and compared for phosphorus (P) removal efficiency and microbial community composition. P-removal efficiency was higher at 20°C (>90%) than at 30°C (60%) when the sludge retention time (SRT) was controlled at 30 days by removing excess sludge equally throughout the sludge bed. Samples analyzed by fluorescent in situ hybridization (FISH) indicated a segregation of biomass over the sludge bed: in the upper part, Candidatus Competibacter phosphatis (glycogen-accumulating organisms--GAOs) were dominant while in the bottom, Candidatus Accumulibacter phosphatis (polyphosphate-accumulating organisms--PAOs) dominated. In order to favour PAOs over GAOs and hence improve P-removal at 30°C, the SRT was controlled by discharging biomass mainly from the top of the sludge bed (80% of the excess sludge), while bottom granules were removed in minor proportions (20% of the excess sludge). With the selective sludge removal proposed, 100% P-removal efficiency was obtained in the reactor operated at 30°C. In the meantime, the biomass in the 30°C reactor changed in color from brownish-black to white. Big white granules appeared in this system and were completely dominated by PAOs (more than 90% of the microbial population), showing relatively high ash content compared to other granules. In the reactor operated at 20°C, P-removal efficiency remained stable above 90% regardless of the sludge removal procedure for SRT control. The results obtained in this study stress the importance of sludge discharge mainly from the top as well as in minor proportions from the bottom of the sludge bed to control the SRT in order to prevent significant growth of GAOs and remove enough accumulated P from the system, particularly at high temperatures (e.g., 30°C).

Effect of different operational conditions on biofilm development, nitrification, and nitrifying microbial population in moving-bed biofilm reactors.

In this study, the effect of different operational conditions on biofilm development and nitrification in three moving-bed biofilm reactors (MBBRs) was investigated: two reactors were operated in a continuously fed regime and one in sequencing-batch mode. The presence of organic carbon reduced the time required to form stable nitrifying biofilms. Subsequent stepwise reduction of influent COD caused a decrease in total polysaccharide and protein content, which was accompanied by a fragmentation of the biofilm, as shown by scanning electron microscopy, and by an enrichment of the biofilm for nitrifiers, as observed by fluorescent in situ hybridization (FISH) analysis. Polysaccharide and protein concentrations proved to be good indicators of biomass development and detachment in MBBR systems. Ammonium- and nitrite-oxidizing bacteria activities were affected when a pulse feeding of 4 g of NH(4)-N/(m(2)·day) was applied. Free nitrous acid and free ammonia were likely the inhibitors for ammonium- and nitrite-oxidizing bacteria.

Effect of elevated salt concentrations on the aerobic granular sludge process: linking microbial activity with microbial community structure.

The long- and short-term effects of salt on biological nitrogen and phosphorus removal processes were studied in an aerobic granular sludge reactor. The microbial community structure was investigated by PCR-denaturing gradient gel electrophoresis (DGGE) on 16S rRNA and amoA genes. PCR products obtained from genomic DNA and from rRNA after reverse transcription were compared to determine the presence of bacteria as well as the metabolically active fraction of bacteria. Fluorescence in situ hybridization (FISH) was used to validate the PCR-based results and to quantify the dominant bacterial populations. The results demonstrated that ammonium removal efficiency was not affected by salt concentrations up to 33 g/liter NaCl. Conversely, a high accumulation of nitrite was observed above 22 g/liter NaCl, which coincided with the disappearance of Nitrospira sp. Phosphorus removal was severely affected by gradual salt increase. No P release or uptake was observed at steady-state operation at 33 g/liter NaCl, exactly when the polyphosphate-accumulating organisms (PAOs), "Candidatus Accumulibacter phosphatis" bacteria, were no longer detected by PCR-DGGE or FISH. Batch experiments confirmed that P removal still could occur at 30 g/liter NaCl, but the long exposure of the biomass to this salinity level was detrimental for PAOs, which were outcompeted by glycogen-accumulating organisms (GAOs) in the bioreactor. GAOs became the dominant microorganisms at increasing salt concentrations, especially at 33 g/liter NaCl. In the comparative analysis of the diversity (DNA-derived pattern) and the activity (cDNA-derived pattern) of the microbial population, the highly metabolically active microorganisms were observed to be those related to ammonia (Nitrosomonas sp.) and phosphate removal ("Candidatus Accumulibacter").

MBBR evaluation for oil refinery wastewater treatment, with post-ozonation and BAC, for wastewater reuse.

This work evaluated the performance of a Moving Bed Biofilm Reactor (MBBR) in the treatment of an oil refinery wastewater. Also, it investigated the possibility of reuse of the MBBR effluent, after ozonation in series with a biological activated carbon (BAC) column. The best performance of the MBBR was achieved with a hydraulic retention time (HRT) of 6 hours, employing a bed to bioreactor volume ratio (V(B)/V(R)) of 0.6. COD and N-NH4(+) MBBR effluent concentrations ranged from 40 to 75 mg L⁻¹ (removal efficiency of 69-89%) and 2 to 6 mg L⁻¹ (removal efficiency of 45-86%), respectively. Ozonation carried out for 15 min with an ozone concentration of 5 mg L⁻¹ was able to improve the treated wastewater biodegradability. The treatment performance of the BAC columns was practically the same for ozonated and non ozonated MBBR effluents. The dissolved organic carbon (DOC) content of the columns of the activated carbon columns (CAG) was in the range of 2.1-3.8 mg L⁻¹, and the corresponding DOC removal efficiencies were comprised between 52 and 75%. The effluent obtained at the end of the proposed treatment presented a quality, which meet the requirements for water reuse in the oil refinery.

Toxicity assessment of oil field produced water treated by evaporative processes to produce water to irrigation.

During the productive life of an oil well, a high quantity of produced water is extracted together with the oil, and it may achieve up to 99% in the end of the well's economical life. Desalination is one of mankind's earliest forms of saline water treatment, and nowadays, it is still a common process used throughout the world. A single-effect mechanical vapor compression (MVC) process was tested. This paper aims to assess the potential toxicity of produced water to be re-used in irrigation. Samples of both produced and distilled water were evaluated by 84 chemical parameters. The distilled produced water presented a reduction up to 97% for the majority of the analyzed parameters, including PAHs. Toxicity bioassays were performed with distilled produced water to evaluate the growth inhibition of Pseudokirchneriella subcapitata algae, the acute toxicity to Danio rerio fish, the germination inhibition of Lactuca sativa vegetable and the severity of toxicity, as well as behavior test with Lumbricid Earthworm Eisenia fetida. The ecotoxicological assays results showed no toxicity, indicating that the referred evaporative process can produce water to be reused in irrigation.

Application of ozonation to reduce biological sludge production in an industrial wastewater treatment plant.

Biosolids production in the activated sludge process generates an additional cost to wastewater treatment plants due to the growing requirements for sludge treatment and disposal. This work focuses on the application of ozonation to reduce sludge production in an industrial wastewater treatment plant. The results show that ozonation was able to promote cell wall rupture, releasing intracellular matter into the liquid medium. This effect was observed by the increase in concentrations of DNA (1.14 to 7.83 mg/L) and proteins (0.5 to 45.602 mg/L) in the liquid phase, when ozonation was applied during 10 min, using 30 mg/L of ozone. Reduction of sludge production was assessed by calculating the observed sludge yield coefficient (Y) in bench-scale continuous experiments conducted with varying proportions of ozonated sludge in the recycle stream and recycle ratios. Reduction of sludge production ranged from 14 to 39%, depending on the experimental conditions. The best result in terms of sludge excess reduction was achieved when 20% of the recycle sludge was ozonated and the recycle ratio was 0.67.

Toxicity evaluation of the process effluent streams of a petrochemical industry.

The physico-chemical characteristics and the acute toxicity of several wastewater streams, generated in the industrial production of synthetic rubber, were determined. The acute toxicity was evaluated in bioassays using different organisms: Danio rerio (fish), Lactuca sativa (lettuce) and Brachionus calyciflorus (rotifer). The removal of toxicity attained in the industrial wastewater treatment plant was also determined upstream and downstream of the activated sludge process. The results obtained indicate that the critical streams in terms of acute toxicity are the effluents from the liquid polymer unit and the spent caustic butadiene washing stage. The biological treatment was able to partially remove the toxicity of the industrial wastewater. However, a residual toxicity level persisted in the biotreated wastewater. The results obtained with Lactuca sativa showed a high degree of reproducibility, using root length or germination index as evaluation parameters. The effect of volatile pollutants on the toxicity results obtained with lettuce seeds was assessed, using ethanol as a model compound. Modifications on the assay procedure were proposed. A strong correlation between the toxic responses of Lactuca sativa and Danio rerio was observed for most industrial effluent streams.

Treatment of the butadiene washing stream from a synthetic rubber industry and recovery of p-TBC.

Butadiene is marketed containing p-tertbutylcatechol (p-TBC), a polymerization inhibitor that should be removed before butadiene utilization in synthetic rubber production. p-TBC can be removed from butadiene by washing with sodium hydroxide (NaOH) solution, producing a wastewater with pH 14, which contains high amounts of p-TBC, a toxic chemical compound. The aim of this work was to develop a treatment process that could reduce the content of p-TBC from the wastewater. Since p-TBC is very soluble in basic, but not in neutral and acid solutions, acidification tests were performed with phosphoric acid (H3PO4) to precipitate p-TBC. Reaction between NaOH and H3PO4 can result in crystallization of large amounts of salt without p-TBC precipitation. Under selected acidifying conditions p-TBC precipitates and the wastewater COD is highly reduced (> 90/o). Chromatographic determinations showed that the precipitated p-TBC could be recovered with 99% purity.

Osteoblastoma of the mandible: systematic review of the literature and report of a case.

Benign osteoblastoma is a bone tumour that seldom occurs in the facial bones. The objective of this article is to add one more case of this rare lesion to the academic literature and to provide a systematic review of previously published cases. A new case of benign osteoblastoma is presented and clinical, radiographic and microscopic aspects, as well as differential diagnosis, treatment and follow-up are discussed. The importance of the correct diagnosis of this type of lesion is stressed, since it presents a clinical, radiographic and microscopic similarity to other bone lesions, including malignant tumours, which may lead the professional into conducting the case in an improper manner.

Phenol removal from high salinity effluents using Fenton's reagent and photo-Fenton reactions.

The removal of pollutants in saline medium by the Fenton's reagent needs a more detailed investigation, since the presence of chloride may inhibit or retard degradation. Phenol was used as a model pollutant and the influence of some important process variables for the removal of total organic carbon and phenol were investigated, such as FeSO4 and H2O2 concentrations, pH and salinity. The reactivity of iron cations and alternative procedures of applying UV radiation (photo-Fenton) were evaluated. Phenol was fast and completely removed by the Fenton's process even in a high saline medium (50,000mg NaCll(-1)). However, TOC was only moderately or poorly removed in saline media, depending on the salt concentration. When the photo-Fenton process was used, mineralization was improved and high TOC removals were observed in moderately saline media (NaCl concentration < or =10,000mgl(-1)). For the highest NaCl concentration tested (50,000mgl(-1)) only a moderate TOC removal was observed (50%).