Hypo-osmotic Stress Increases Permeability of Individual Barriers in Escherichia coli Cell Envelope, Enabling Rapid Drug Transport.

Survival of foodborne Gram-negative bacteria during osmotic stress often leads to multidrug resistance development. However, despite the concern, how osmoadaptation alters drug penetration across the Gram-negative bacterial cell envelope has remained inconclusive for years. Here, we have investigated drug permeation and accumulation inside hypo-osmotically shocked Escherichia coli. Three different quaternary ammonium compounds (QACs) are used as cationic amine-containing drug representatives; they also serve as envelope permeability indicators in different assays. Propidium iodide fluorescence reveals cytoplasmic accumulation and overall envelope permeability, while crystal violet sorption and second harmonic generation (SHG) spectroscopy reveal periplasmic accumulation and outer membrane permeability. Malachite green sorption and SHG results reveal transport across both the outer and inner membranes and accumulation in the periplasm as well as cytoplasm. The findings are found to be complementary to one another, collectively revealing enhanced permeabilities of both membranes and the periplasmic space in response to hypo-osmotic stress in E. coli. Enhanced permeability leads to faster QACs transport and higher accumulation in subcellular compartments, whereas transport and accumulation both are negligible under isosmotic conditions. The QACs' transport rates are found to be highly influenced by the osmolytes used, where phosphate ion emerges as a key facilitator of transport across the periplasm into the cytoplasm. E. coli is found viable, with morphology unchanged under extreme hypo-osmotic stress; i.e., it adapts to the situation. The outcome shows that the hypo-osmotic shock to E. coli, specifically using phosphate as an osmolyte, can be beneficial for drug delivery.

Deep eutectic solvent-based liquid-phase microextraction coupled with reversed-phase high-performance liquid chromatography for determination of α-, ß-, γ-, and δ-tocopherol in edible oils.

For simultaneous analysis of four fat-soluble tocopherols (α-, ß-, γ-, and δ-) in edible oils, an efficient and green method using deep eutectic solvent-based liquid-phase microextraction (DES-LPME) coupled with reversed-phase high-performance liquid chromatography (RP-HPLC) was developed. The DESs formed by different quaternary ammonium salts and ethanol were used as the extractants. Tetrabutylammonium chloride (TBAC)-ethanol DES at a molar ratio of 1:2 achieved the best extraction efficiency. Under the optimized conditions, the detection limits were in the range of 2.1-3.0 ng mL-1. The intra-day and inter-day repeatability were in the ranges of 3.9-5.3% and 4.8-7.1%, respectively, and the recoveries for the real samples varied from 80.7% to 105.4%. The developed method was successfully employed for the determination of all four tocopherol homologues with an RP-HPLC system containing a COSMOSIL π-NAP column in five edible oils collected locally. Graphical abstract.

Fluorometric assay of iron(II) lactate hydrate and ammonium ferric citrate in food and medicine based on poly(sodium-p-styrenesulfonate)-enhanced Ag nanoclusters.

Poly(sodium-p-styrenesulfonate)-enhanced and D-penicillamine stabilized Ag nanoclusters (PSS-DPA-AgNCs) were prepared using one-step ultraviolet irradiation combined with microwave heating method, and the effects of the AgNCs photo-luminescence performance based on different types of polyelectrolytes and energy suppliers were studied detailedly. The as-prepared AgNCs can be used as a viable fluorescent probe for monitoring indirectly iron(II) lactate hydrate (ILH) and ammonium ferric citrate (AFC), respectively. The fluorescence (FL) quenching of PSS-DPA-AgNCs by Fe3+ (it is obtained from oxidized ILH/ionized AFC) mainly derives from a dynamic quenching process. Excellent linear relationships exist between the FL quenching degree of the AgNCs and the concentrations of ILH/AFC in the range of 0.17-6.00/0.067-3.33 µmol·L-1, and corresponding limit of detection (at 3σ/slope) is 12.4/6.04 nmol·L-1. Moreover, the AgNCs probe was extended to the assays of ILH in tablets, solid beverage or ILH additive and AFC in two kinds of edible salts or syrup with satisfactory results compared with the standard 1, 10-phenanthroline method. In addition, the AgNCs probe reveals a good temperature sensing capability.

Multistep approach to control microbial fouling of historic building materials by aerial phototrophs.

The scientific multistep approach described herein is a result of two years of research into a control method against microbial fouling and biodeterioration of historic building materials by phototrophs. A series of tests were conducted to select the best antifouling agent for eliminating 'green' coatings and protecting surfaces against biofouling. Of the seven active compounds, two with the best penetration abilities were subjected to a photosynthetic activity inhibition test using confocal microscopy. Of the two, a quaternary ammonium salt (QAC) - didecyldimethylammonium chloride (DDAC) - was found to be the most effective. Ten biocides containing QACs at different concentrations were then tested against 'green' coatings on wood, brick and plaster, with the best four being selected for further research in model conditions. As a result, biocides containing >14% (v v-1) DDAC were found to be successful antifouling agents for protecting historical materials against biodeterioration by phototrophs.

Modeling quaternary ammonium compound inhibition of biological nutrient removal activated sludge.

Quaternary ammonium compounds (QACs) are surface-active organic compounds common in industrial cleaner formulations widely used in various sanitation applications. While acting as effective pathogenic biocides, QACs lack selective toxicity and often have poor target specificity. As a result, adverse effects on biological processes and thus the performance of biological nutrient removal (BNR) systems may be encountered when QACs enter wastewater treatment plants (WWTPs). Because of these impacts, there is motivation to screen wastewater influents for QACs and for process engineers to consider the inhibition effects of QACs on process evaluation and design of BNR plants. This paper introduces a mathematical model to describe the fate of QACs in a WWTP via biodegradation and bio-adsorption, and the inhibitory effect of QACs on nitrifiers and ordinary heterotrophic organisms. The model was incorporated as an add-on model in BioWin 5.3 and simulations of experimental systems were used for comparison of model results to measured data reported in the literature. The model was found to accurately predict the bulk phase concentration of QAC and the inhibition of nitrification with QAC concentrations ≥2 mg/L. This work provides a preliminary framework for simulation of BNR plants receiving inhibitory substances in the influent.

Quaternary ammonium compounds in roots and leaves of Capparis spinosa L. from Saudi Arabia and Italy: investigation by HPLC-MS and 1H NMR.

Capparis spinosa L. is a perennial plant typical of the Mediterranean flora and a multipurpose plant used for curing various human ailments. Quaternary ammonium compounds (QACs), as constituents of Capparaceae, play important roles in protecting against abiotic stress. Aim of this work was to determine QACs in root and leaves of caper from two proveniences. The presence of stachydrine, choline, glycine betaine and homo-stachydrine has been confirmed by high resolution MS, while 1H NMR was applied to quantify the main QACs in the aqueous extracts. Stachydrine was quantified at 20.2 mg/g and 32.3 mg/g on dry leaves from South of Italy and Saudi Arabia, respectively, while a minor content was in dry roots (from 10.4 to 12.5 mg/g). Choline was considerably lower both in leaves and roots (from 0.3 to 1.2 mg/g). To our knowledge, this is the first report on the determination of QACs both in root and leaves of C. spinosa.

Isolation of Vacuoles from the Leaves of the Medicinal Plant Catharanthus roseus.

The isolation of vacuoles is an essential step to unravel the important and complex functions of this organelle in plant physiology. Here, we describe a method for the isolation of vacuoles from Catharanthus roseus leaves involving a simple procedure for the isolation of protoplasts, and the application of a controlled osmotic/thermal shock to the naked cells, leading to the release of intact vacuoles, which are subsequently purified by density gradient centrifugation. The purity of the isolated intact vacuoles is assayed by microscopy, western blotting, and measurement of vacuolar (V)-H+-ATPase hydrolytic activity. Finally, membrane functionality and integrity is evaluated by measuring the generation of a transtonoplast pH gradient by the V-H+-ATPase and the V-H+-pyrophosphatase, also producing further information on vacuole purity.

[Effect of NH4(+) -N/NO3(-)-N ratio in applied supplementary fertilizer on nitrogen metabolism and main chemical composition of Pinellia ternata].

OBJECTIVE: To study the effect of nitrogen forms on nitrogen metabolism and main chemical composition of Pinellia ternate. METHOD: Through the soilless cultivation experiment and based at the same nitrogen level and different NH4(+) -N/NO3(-) -N ratios, nitrate reductase (NR) activity, glutamine synthetase (GS) activity, the content of nitrate nitrogen and ammonium nitrogen in different parts of P. ternate were determined. The contents of total alkaloid, free total organic acids and guanosine in the tuber were determined. The yield of bulbil and tuber was calculated. RESULT: The test results showed that, with the NH4(+) -N/NO3(-) -N ratio increasing, the activity of nitrate reductase decreased, the content of nitrate nitrogen in the leaves, petioles and tuber increasing initially, then decreased, and the content of nitrate nitrogen in the root decreased. Meanwhile, with the NH4(+) -N/NO3(-) -N ratio increasing, the activity of glutamine synthetase in the leaves, petioles and root increased, the activity of glutamine synthetase in the tuber increasing initially, then decreased. The contents of ammonium nitrogen in the leaves, tuber and root increased initially, then decreased, and the contents of ammonium nitrogen in the petioles increased with the NH4(+)(-N/NO3(-)-N ratio increasing. The yield of bulbil and tuber were the highest at the NH4(+)-N/NO3(-) -N ratio of 75: 25. The content of total alkaloid and guanosine in the tuber were the highest at the NH4(+)-N/NO3(-) -N ratio of 0: 100, and the contents were 0.245% and 0.0197% respectively. With the NH4(+)-N/NO3(-) -N ratio of 50: 50, the content of free total organic acids was the highest, it reached 0.7%, however, the content of free total organic acids was the lowest at the NH4(+) -N/NO3(-) -N ratio of 0: 100. CONCLUSION: Nitrogen fertilization significant influences the nitrogen metabolism, the yield and main chemical composition of P. ternate.

Seasonal variations and aeration effects on water quality improvements and physiological responses of Nymphaea tetragona Georgi.

Seasonal variations and aeration effects on water quality improvements and the physiological responses of Nymphaea tetragona Georgi were investigated with mesocosm experiments. Plants were hydroponically cultivated in six purifying tanks (aerated, non-aerated) and the characteristics of the plants were measured. Water quality improvements in purifying tanks were evaluated by comparing to the control tanks. The results showed that continuous aeration affected the plant morphology and physiology. The lengths of the roots, petioles and leaf limbs in aeration conditions were shorter than in non-aeration conditions. Chlorophyll and soluble protein contents of the leaf limbs in aerated tanks decreased, while peroxidase and catalase activities of roots tissues increased. In spring and summer, effects of aeration on the plants were less than in autumn. Total nitrogen (TN) and ammonia nitrogen (NH4(+)-N) in aerated tanks were lower than in non-aerated tanks, while total phosphorus (TP) and dissolved phosphorus (DP) increased in spring and summer. In autumn, effects of aeration on the plants became more significant. TN, NH4(+)-N, TP and DP became higher in aerated tanks than in non-aerated tanks in autumn. This work provided evidences for regulating aeration techniques based on seasonal variations of the plant physiology in restoring polluted stagnant water.

Enhancement of phosphorus removal in a low temperature A(2)/O process by anaerobic phosphorus release of activated sludge.

An anaerobic phosphorus release tank was introduced to an anaerobic-anoxic-aerobic (A(2)/O) process treating domestic sewage to enhance the phosphorus removal at low temperature. Phosphorus release of the activated sludge from the second sedimentation tank was evaluated at 14 °C by batch cultures, and the nutrient removal in the modified low temperature A(2)/O process was further investigated at the same temperature. The results showed that the feasible sludge retention time was 14 h for sequencing batch reaction and 12 h for continuous flow operation. The ratio of raw sewage to activated sludge from the second sedimentation tank was 1:1 in volume to meet the demand of carbon resource for the growth of phosphorus release microbes. The feasible chemical oxygen demand (COD) loading rate of the activated sludge in the phosphorus release tank was 0.015-0.02 g COD/g MLSS (mixed liquor suspended solids) and the nitrate concentration should be less than 5 mg/L. The phosphorus release was doubled when the sludge was blended intermittently and gently. The anaerobic phosphorus release of the activated sludge improved the phosphate removal remarkably, as well as the removal of NH4(+)-N and total nitrogen (TN) in the modified low temperature A(2)/O process. The effluent COD, NH4(+)-N, TN and total phosphorus could meet a stricter discharge standard.

The influence of Castanhão reservoir on nutrient and suspended matter transport during rainy season in the ephemeral Jaguaribe river (CE, Brazil).

Measurements of nutrient and suspended matter concentrations and loads entering and leaving the Castanhão reservoir during the rainy season were carried out to assess the influence of this large reservoir on land-sea fluvial transport in the ephemeral Jaguaribe river basin. Spatial variation indicated statistically significant attenuation of concentrations only for total phosphorous and suspended matter across the reservoir. Strong retention of nutrients and suspended matter loads by the reservoir was observed with average trapping efficiency of 89% for dissolved silicon, 98% of soluble reactive phosphorus, 71% for ammonium, 87% for total nitrogen, 98% for total phosphorus and 97% for suspended matter compared to the reservoir inflow. The dam operational procedure defined by the ephemeral conditions of the river reduced water releases compared to reservoir inflow and induced strong retention of nutrient and suspended matter loads within the reservoir when fluvial transfer occurs in this semiarid watershed.

In situ analysis of corrosion inhibitors using a portable mass spectrometer with paper spray ionization.

Paper spray (PS) ambient ionization is implemented using a portable mass spectrometer and applied to the detection of alkyl quaternary ammonium salts in a complex oil matrix. These salts are commonly used as active components in the formulation of corrosion inhibitors. They were identified in oil and confirmed by their fragmentation patterns recorded using tandem mass spectrometry (MS/MS). The cations of alkyl and benzyl-substituted quaternary ammonium salts showed characteristic neutral losses of CnH2n (n carbon number of the longest chain) and C7H8, respectively. Individual quaternary ammonium compounds were detected at low concentrations (<1 ng µL(-1)) and over a dynamic range of ∼5 pg µL(-1) to 500 pg µL(-1) (ppb). Direct detection of these compounds in complex oil samples without prior sample preparation or pre-concentration was also demonstrated using a home-built miniature mass spectrometer at levels below 1 ng µL(-1).

Caragana fabr. promotes revegetation and soil rehabilitation in saline-alkali wasteland.

To determine how plantations of Caragana microphylla shrubs affect saline-alkali soil amelioration and revegetation, we investigated the vegetation and sampled soils from saline-alkali wasteland (SAW), perennial Caragana forestland (PCF), Caragana forest after fire disturbance (CFF). Results showed that with the development of Caragana Fabr., highly dominant species of Poaceae family, including Elymus dahuricus, Thermopsis lanceolata, Stipa tianschanica, died out in PCF. Moreover, Papilionaceaefamily, including Lespedeza indica, Oxytropis psammocharis, and Astragalus scaberrimus, was established both in PCF and CFF. Phytoremediation of saline-alkali wasteland (SAW) was achieved by plantation, resulting in the reduced soil pH, sodium adsorption ratio, exchangeable sodium percentage, salinity, and Na+ concentration around Caragana shrubs. Greater amounts of soil organic, total nitrogen, ammonium nitrogen, available phosphorus, and available potassium were observed in PCF topsoil than in SAW topsoil The concentration of mineralized N in PCF soil was significantly lower than that in SAW soil at all sampled depths, indicating that Caragana shrubs were just using N and therefore less measured in soils. Fire disturbance resulted in decreased soil pH and salinity, but increased organic content, total nitrogen, and ammonium nitrogen. The improved soil parameters and self-recovery of shrubs indicated that Caragana shrubs were well established after burning event.

Interaction between nitrogen and phosphorus cycles in mining-affected aquatic systems-experiences from field and laboratory measurements.

The main objectives of this study were to (a) study the interaction between N and P cycles in mining-affected aquatic systems and (b) to quantify release rates of sedimentary soluble reactive phosphorus (SRP) that may be related to this interaction. Sediment cores and water from Lake Bruträsket (Boliden, northern Sweden) were collected and a time series of water sampling and flow measurements was conducted in the Brubäcken stream connected to the lake. Factors affecting SRP release were studied in a sediment incubation experiment and water column experiments. Field and laboratory measurements indicated that pH and dissolved oxygen are two important factors for SRP release. At the end of the low-oxygen incubation, an SRP concentration of 56 µg L(-1) resulted in a sedimentary flux of 1.1 mg SRP m(-2) day(-1). This is ~10 times higher than the flux of 0.12 mg SRP m(-2) day(-1) obtained from depth integration of vertical SRP profiles measured in the lake, and ~100 times higher than the external flux of 0.014 mg SRP m(-2) d(-1) into the lake (based on catchment area). Field measurements indicated that oxidation of organic matter and mining-related chemicals (ammonium and thiosulphates) may result in increased internal SRP flux from the sediment. Increased P loading in the lake as a result of low-oxygen conditions could change water column total nitrogen/total phosphorus ratios from 27 to 17, consequently changing the lake from being P-limited to be co-limited by N and P. The obtained findings point to possible interaction between the cycles of nitrogen (oxygen consumption) and P (flux from sediment) that may be important for nutrient regulation in mine water recipients.

Nitrogen cycle feedbacks as a control on euxinia in the mid-Proterozoic ocean.

Geochemical evidence invokes anoxic deep oceans until the terminal Neoproterozoic ~0.55 Ma, despite oxygenation of Earth's atmosphere nearly 2 Gyr earlier. Marine sediments from the intervening period suggest predominantly ferruginous (anoxic Fe(II)-rich) waters, interspersed with euxinia (anoxic H(2)S-rich conditions) along productive continental margins. Today, sustained biotic H(2)S production requires NO(3)(-) depletion because denitrifiers outcompete sulphate reducers. Thus, euxinia is rare, only occurring concurrently with (steady state) organic carbon availability when N(2)-fixers dominate the production in the photic zone. Here we use a simple box model of a generic Proterozoic coastal upwelling zone to show how these feedbacks caused the mid-Proterozoic ocean to exhibit a spatial/temporal separation between two states: photic zone NO(3)(-) with denitrification in lower anoxic waters, and N(2)-fixation-driven production overlying euxinia. Interchange between these states likely explains the varying H(2)S concentration implied by existing data, which persisted until the Neoproterozoic oxygenation event gave rise to modern marine biogeochemistry.

The influence of castanhão reservoir on nutrient and suspended matter transport during rainy season in the ephemeral Jaguaribe river (CE, Brazil) / Influência do açude Castanhão no transporte de nutrientes e materiais em suspensão durante a estação chuvosa no intermitente rio Jaguaribe (CE, Brasil)

Measurements of nutrient and suspended matter concentrations and loads entering and leaving the Castanhão reservoir during the rainy season were carried out to assess the influence of this large reservoir on land-sea fluvial transport in the ephemeral Jaguaribe river basin. Spatial variation indicated statistically significant attenuation of concentrations only for total phosphorous and suspended matter across the reservoir. Strong retention of nutrients and suspended matter loads by the reservoir was observed with average trapping efficiency of 89% for dissolved silicon, 98% of soluble reactive phosphorus, 71% for ammonium, 87% for total nitrogen, 98% for total phosphorus and 97% for suspended matter compared to the reservoir inflow. The dam operational procedure defined by the ephemeral conditions of the river reduced water releases compared to reservoir inflow and induced strong retention of nutrient and suspended matter loads within the reservoir when fluvial transfer occurs in this semiarid watershed.

Medidas de concentrações e fluxos de nutrientes e material particulado em suspensão de entrada e saída do Açude Castanhão foram efetuadas para determinar a influência de um grande reservatório no transporte fluvial entre o continente e o oceano na bacia hidrográfica intermitente do Rio Jaguaribe. A variação espacial indicou uma atenuação estatisticamente significativa ao longo do reservatório somente para as concentrações de fósforo total e material particulado em suspensão. Os resultados indicam uma forte retenção de nutrientes e do material particulado em suspensão com eficiências de retenção de 89% para sílica dissolvida, 98% para fósforo solúvel reativo, 71% para amônia, 87% para nitrogênio total, 98% para fósforo total e 97% para o material particulado em suspensão em relação aos fluxos de entrada no reservatório. A rotina de operação da barragem definida pela condição intermitente do Rio Jaguaribe reduz a descarga de água comparada com a entrada no reservatório e induz forte retenção do fluxo fluvial de nutrientes e material particulado em suspensão pelo reservatório.

Reclamation of cadmium-contaminated soil using dissolved organic matter solution originating from wine-processing waste sludge.

Soil washing using an acid solution is a common practice for removing heavy metals from contaminated soil in Taiwan. However, serious loss of nutrients from soil is a major drawback of the washing. Distillery sludge can be used to prepare a dissolved organic matter (DOM) solution by extracting its organic constituents with alkaline solutions. This study employed DOM solutions to remediate Cd-contaminated soil (with concentrations up to 21.5 mg kg(-1)) and determine the factors affecting removal of Cd, such as pH, initial concentration of DOM solution, temperature, and washing frequency. When washing with pH 3.0 and 1250 mg L(-1) DOM solution, about 80% and 81% of Cd were removed from the topsoil at 27 °C and subsoil at 40 °C, respectively. To summarize the changes in fertility during DOM washing with various pH solutions: the increase in organic matter content ranged from 7.7% to 23.7%; cation exchange capacity (CEC) ranged from 4.6% to 13.9%; available ammonium (NNH(4)) content ranged from 39.4% to 2175%; and available phosphorus content ranged from 34.5% to 182%. Exchangeable K, Ca, and Mg remained in the topsoil after DOM washing, with concentrations of 1.1, 2.4, and 1.5 times higher than those treated with HCl solution at the same pH, respectively.

Performance evaluation of different horizontal subsurface flow wetland types by characterization of flow behavior, mass removal and depth-dependent contaminant load.

For several pilot-scale constructed wetlands (CWs: a planted and unplanted gravel filter) and a hydroponic plant root mat (operating at two water levels), used for treating groundwater contaminated with BTEX, the fuel additive MTBE and ammonium, the hydrodynamic behavior was evaluated by means of temporal moment analysis of outlet tracer breakthrough curves (BTCs): hydraulic indices were related to contaminant mass removal. Detailed investigation of flow within the model gravel CWs allowed estimation of local flow rates and contaminant loads within the CWs. Best hydraulics were observed for the planted gravel filter (number of continuously stirred tank reactors N = 11.3, dispersion number = 0.04, Péclet number = 23). The hydroponic plant root mat revealed lower N and pronounced dispersion tendencies, whereby an elevated water table considerably impaired flow characteristics and treatment efficiencies. Highest mass removals were achieved by the plant root mat at low level: 98% (544 mg m⁻² d⁻¹), 78% (54 mg m⁻² d⁻¹) and 74% (893 mg m⁻² d⁻¹) for benzene, MTBE and ammonium-nitrogen, respectively. Within the CWs the flow behavior was depth-dependent, with the planting and the position of the outlet tube being key factors resulting in elevated flow rate and contaminant flux immediately below the densely rooted porous media zone in the planted CW, and fast bottom flow in the unplanted reference.

Sediment-seawater solute flux in a polluted New Zealand estuary.

We investigated the sediment-seawater solute flux at five sites in the polluted Avon-Heathcote Estuary, New Zealand, to provide a point of comparison for future studies of the effects of the closure of a major wastewater outfall into the estuary. Sediments collected in winters 2007 and 2008, and summer 2008, ranked consistently in organic matter content. Microelectrode profiling and sediment-core incubations revealed (1) a dominant role of microphytes in regulating solute flux causing significant differences in the dark and light sediment O2 consumption (R(d), R(l)), total sediment O2 utilisation (TOU(d), TOU(l)), and inorganic nutrient flux, (2) consistent ranking of sites in solute flux, and (3) a clear solute-flux signature of the wastewater effluent. Sediment near the wastewater outfall exhibited the highest absolute R and TOU(,) the lowest ratio R(l)/R(d,) the highest dark efflux of dissolved reactive phosphorus and ammonium, and the highest dark and light uptake of nitrate+nitrite.

Performance of intermittently aerated up-flow sludge bed reactor and sequencing batch reactor treating industrial estate wastewater: a comparative study.

In this study, an innovative aerobic/anoxic sludge bed bioreactor with two feeding regimes, continuous-fed (an up-flow sludge bed reactor (USBR)) and batch fed (sequencing batch reactor (SBR)), was evaluated for the treatment of an industrial estate wastewater with low BOD(5)/COD ratio. The process performance in the two regimes was compared. Two numerical independent variables (retention/react time and aeration time) were selected to analyze, model and optimize the process. Response surface methodology with central composite design (CCD) was used with five levels of hydraulic retention time (HRT)/react time (12-36h) and aeration time (40-60min/h). In order to analyze the process, ten dependent parameters as the process responses were studied. As a result, HRT/react time showed a decreasing impact on the responses measured in both hydraulic regimes, USBR and SBR. The USBR showed better performance than the SBR in removal of total COD, slowly biodegradable COD, total nitrogen and total Kjeldahl nitrogen.