Removal capacity of water plant alum sludge for phosphorus in aqueous solutions

Abstract

Alum sludge, which is a waste product from a potable water treatment process, was tested as an inexpensive alternate adsorbent for phosphorus in wastewater. The sludge was composed dominantly of sand size aggregates, and could remain stable in aqueous media. The majority of reactive Al in alum sludge was present as an amorphous phase, and seemed to be the major absorbent for P. The batch sorption test showed that the removal of P was influenced by the solubility of Al, Fe and organic carbon depending on pH condition. The acidic condition favored the removal of P, and there was a side effect in the P removal process such as dissolution of Al and organic C at acidic (pH < 4) and alkaline (pH < 8) conditions. The pH range from 4 to 6 was effective for all inorganic and organic phosphates with a low solubility of Al and organic C. The maximum adsorption capacity of alum sludge was calculated as 25,000 mg/kg for orthophosphate, and followed the order: orthophosphate > pyrophosphate > triphosphate > organic phosphate (adenosin). From the column test with a 30 mg/L orthophosphate solution at a flow rate of 3.0 ml/min, the alum sludge removed P to less than 1.0 mg/L over 250 pore volumes at initial pH 4, and 200 pore volumes at initial pH 5, respectively.     

Biological Uptake of Phosphorus by Activated Sludge

The ability of activated sludge to remove phosphates was studied by adding carrier-free (32)P to raw sewage and measuring incorporation of the radioactivity into the cells over a period of time. Radioisotope determinations indicated that 48% of the (32)P radioactivity was removed by 12 hr. However, chemical methods indicated that only 30% of the orthophosphate apparently disappeared from the sewage during this period. Experiments with sludge prelabeled with (32)P indicated that considerable phosphate turnover occurred. The cells released large amounts of radioactivity as they were incorporating fresh phosphates. Starvation in isotonic saline for 18 hr caused the sludge to dump phosphate. When introduced into fresh sewage containing (32)P, the starved sludge removed about 60% of the radioactivity in 6 hr with little phosphate turnover. The ability of sludge to remove (32)P was inhibited approximately 83% by 10(-3)m 2,4-dinitrophenol. This inhibition was at the expense of the cell fraction that contained ribonucleic acid and deoxyribonucleic acid. The sludge cells released orthophosphate when exposed to the chemical agent. Experiments using (45)Ca indicated that calcium phosphate precipitation plays a minor role in phosphate removal under our experimental conditions.     

Culture of Scenedesmus sp. LX1 in the modified effluent of a wastewater treatment plant of an electric factory by photo-membrane bioreactor

To investigate the coupled technology for advanced wastewater treatment and microalgal biomass production, a photo-membrane bioreactor was constructed. The microalga Scenedesmus sp. LX1 was cultured in the bioreactor using liquor prepared from the effluent of an electronic device factory. The algal cell growth, nitrate nitrogen removal, orthophosphate phosphorus removal were investigated. When cultured with batch operation, the average specific growth rate was about 0.09 d⁻¹, and low nitrogen (N), phosphorus (P) concentrations in the liquor were achieved. However, under continuous operation with an inflow of 60 L h⁻¹, the average specific growth rate was only 0.02 d⁻¹, and removal rates of 100% for orthophosphate P and 46% for nitrate N were achieved. With the inflow of 120 L h⁻¹, the accumulated metal ions in the bioreactor adversely affected the algal cells. The algal cells were much easier to settle, and the removal efficiency for N and P decreased.     

Effect of aspartate and glutamate on the fate of enhanced biological phosphorus removal process and microbial community structure

This study investigated the fate of enhanced biological phosphorus removal (EBPR) and changes in microbial speciation in a sequencing batch reactor (SBR) fed with aspartate and glutamate. It involved SBR operation for 288 days, batch tests for observation of metabolic functions together with microscopic and phylogenetic analyses. Polyphosphate accumulating organisms (PAOs) were observed in abundance with complete removal of phosphorus. Fluorescence in situ hybridization (FISH) combined with 4′,6-dia-midino-2-phenylindole (DAPI) staining confirmed the accumulation of polyphosphate by Rhodocyclus-related and Actinobacterial PAOs. Aspartate seemed to favor the competitive growth of Rhodocyclus-related PAOs since EBPR population used the common biochemical pathways followed by Rhodocyclus-related PAOs in the aspartate fed batch tests. In the glutamate fed batch reactors, however, Actinobacterial PAOs appeared to be competitively selected which explains the lower levels of PHA generation. Even though operational conditions did not change, effective EBPR could not be maintained during the latter part of the study.     

Involvement of glycolate metabolism in acclimation of Chlorella vulgaris cultures to low phosphate supply

Chlorella vulgaris (Beijer.) was grown for 8 d under air in cultures with complete (Control) or with phosphorus-deficient (–P) medium limiting culture growth. The cells assimilated only 5–17 % of orthophosphate supplied from the complete medium, whereas from medium of –P cultures, orthophosphate was almost totally exhausted. Despite limited phosphorus availability, cells in the oldest –P cultures contained the same amount of inorganic orthophosphate as the control cells and only slightly less organic phosphates. The –P cells showed normal chlorophyll concentration and increased V_max and 1/K_0.5 dissolved inorganic carbon (DIC) of photosynthetic O₂ evolution. Phosphorus deficiency enhanced production, excretion and metabolism of glycolate during the whole investigated period. In the initial phase of –P culture growth, medium acidification and low DIC concentration were conducive to glycolate production. With subsequent medium alkalization, DIC content and cell carbonic anhydrase activity increased the photosynthetic O₂ evolution of –P cells two-fold. At that period, the elevated intrachloroplast O₂ concentration might be the main reason of enhancement of glycolate metabolism. The results support the suggestion that involvement of glycolate metabolism in acclimation to low phosphorus supply improves regeneration of inorganic orthophosphate and protects chloroplasts against photoinhibitory damage by consumption of excess of absorbed light energy.     

Contaminant removal efficiency depending on primary treatment and operational strategy in horizontal subsurface flow treatment wetlands

This study aimed to evaluate the contaminant removal efficiency of shallow horizontal subsurface flow treatment wetlands (SSF TWs) as a function of (1) primary treatment (hydrolytic upflow sludge blanket (HUSB) reactor vs. conventional settling) and (2) operation strategy (alternation of saturated/unsaturated phases vs. permanently saturated). An experimental plant was constructed, operated and surveyed for the main water quality parameters over a period of 2.5 years. The plant had 3 treatment lines: a control line (settler-wetland permanently saturated), a batch line (settler-wetland operated with saturated/unsaturated phases) and an anaerobic line (HUSB reactor-wetland permanently saturated). In each line wetlands had a surface area of 2.80 m², a water depth of 25 cm and a granular medium D₆₀ = 7.3 mm, and were planted with common reed. During the study period the wetlands were operated at a hydraulic and organic load of 28.5 mm/d and about 4.7 g BOD/m² d, respectively. Effluent average redox potential was lower for the anaerobic line (−45 ± 78 mV) than for the other two lines (3 ± 92.7 and −5 ± 71 mV for control and batch, respectively). Overall, chemical oxygen demand (COD), biochemical oxygen demand (BOD₅) and ammonium mass removal efficiencies were slightly greater for the batch line (88%, 96% and 87%, respectively) than for the control line (83%, 94% and 80%) and the anaerobic line (80%, 87% and 73%). During cold seasons, COD and ammonium removal in the batch line was around 30% and 50% higher than in the control line, respectively. The results of this study indicate that the implementation of a HUSB reactor as primary treatment did not enhance the treatment capacity of the system (in comparison with a conventional settler). The efficiency of treatment wetland systems with horizontal subsurface flow can be improved using a batch operation strategy.     

Performance of laboratory-scale constructed wetlands coupled with micro-electric field for heavy metal-contaminating wastewater treatment

This study reports the performance of laboratory-scale constructed wetlands coupled with micro-electric field (CWMEF) planted cannas (Canna generalis) for heavy metal-contaminating wastewater treatment. The CWMEF had a better performance for heavy metal (HM) removal from wastewater than did the ordinary constructed wetlands (CWs). Owing to the stimulation of the suitable voltage and electrical exposure time, cannas may grew better and in fact assimilated more metallic ions in CWMEF than in CWs. The environmental conditions in CWMEF, such as the higher pH by electrolysis of water, the presence of aluminum ions by anodizing of aluminum, caused chemical precipitation, physical adsorption and flocculation of metallic ions.     

Purification and Characterization of a Phosphoenolpyruvate Phosphatase from Brassica nigra Suspension Cells

Phosphoenolpyruvate phosphatase from Brassica nigra leaf petiole suspension cells has been purified 1700-fold to apparent homogeneity and a final specific activity of 380 micromole pyruvate produced per minute per milligram protein. Purification steps included: ammonium sulfate fractionation, S-Sepharose, chelating Sepharose, concanavalin A Sepharose, and Superose 12 chromatography. The native protein was monomeric with a molecular mass of 56 kilodaltons as estimated by analytical gel filtration. The enzyme displayed a broad pH optimum of about pH 5.6 and was relatively heat stable. Western blots of microgram quantities of the final preparation showed no cross-reactivity when probed with rabbit polyclonal antibodies prepared against either castor bean endosperm cytosolic pyruvate kinase, or sorghum leaf phosphoenolpyruvate carboxylase. The final preparation exhibited a broad substrate selectivity, showing high activity toward p-nitrophenyl phosphate, adenosine diphosphate, adenosine triphosphate, gluconate 6-phosphate, and phosphoenolpyruvate, and moderate activity toward several other organic phosphates. Phosphoenolpyruvate phosphatase possessed at least a fivefold and sixfold greater affinity and specificity constant, respectively, for phosphoenolpyruvate (apparent Michaelis constant = 50 micromolar) than for any other nonartificial substrate. The enzyme was activated 1.7-fold by 4 millimolar magnesium, but was strongly inhibited by molybdate, fluoride, zinc, copper, iron, and lead ions, as well as by orthophosphate, ascorbate, glutamate, aspartate, and various organic phosphate compounds. It is postulated that phosphoenolpyruvate phosphatase functions to bypass the adenosine diphosphate dependent pyruvate kinase reaction during extended periods of orthophosphate starvation.     

Activation of sea urchin eggs by inositol phosphates is independent of external calcium.

We investigated the contribution of external calcium ions to inositol phosphate-induced exocytosis in sea urchin eggs. We show that: (a) inositol phosphates activate eggs of the sea urchin species Lytechinus pictus and Lytechinus variegatus independently of external calcium ions; (b) the magnitude and duration of the inositol phosphate induced calcium changes are independent of external calcium; (c) in calcium-free seawater, increasing the volume of inositol trisphosphate solution injected decreased the extent of egg activation; (d) eggs in calcium-free sea water are more easily damaged by microinjection; microinjection of larger volumes increased leakage from eggs pre-loaded with fluorescent dye. We conclude that inositol phosphates do not require external calcium ions to activate sea urchin eggs. This is entirely consistent with their role as internal messengers at fertilization. The increased damage caused to eggs in calcium-free seawater injected with large volumes may allow the EGTA present in the seawater to enter the egg and chelate any calcium released by the inositol phosphates. This may explain the discrepancy between this and earlier reports.     

The layer effect in nutrient removal by two indigenous plant species in horizontal flow constructed wetlands

A layer effect study was conducted to investigate the transformation of nitrogenous pollutants in two batch subsurface horizontal constructed wetlands. Artificial drainage water containing a low concentration of chemical oxygen demand (COD_Cr), but high concentration of ammonia and nitrate, was treated in two batch wetland cuboids. The nitrogen removal rates were found to be significantly affected by the characters of the layer as well as the biomass and roots of different plant species (P < 0.05). Correlations between pH, oxidation-reduction potential, and retention time indicated that nitrogen removal rates under study conditions mainly depended on the location of the layer and the plant species in the constructed wetland.     

A three-stage experimental constructed wetland for treatment of domestic sewage: First 2 years of operation

Hybrid constructed wetland systems have recently been used to treat wastewaters where high demand for removal of ammonia is required. However, these systems have not been used too often for small on-site treatment systems. This is because in many countries ammonia is not limited in the discharge from small systems. Hybrid systems have a great potential to reduce both ammonia and nitrate concentrations at the same time. In our study we employed a three-stage constructed wetland system consisting of saturated vertical-flow (VF) bed (2.5 m², planted with Phragmites australis), free-drained VF bed (1.5 m², planted with P. australis) and horizontal-flow (HF) bed (6 m², planted with Phalaris arundinacea) in series. All wetlands were originally filled with crushed rock (4-8 mm). However, nitrification was achieved only after the crushed rock was replaced with sand (0-4 mm) in the free-drain wetland. Also, original size of crushed rock proved to be too vulnerable to clogging and therefore, in the first wetlands the upper 40 cm was replaced by coarser fraction of crushed rock (16-32 mm) before the second year of operation started. The system was fed with mechanically pretreated municipal wastewater and the total daily flow was divided into two batches 12 h apart. The evaluation of the results from the period 2007 to 2008 indicated that such a system has a great potential for oxidation of ammonia and reduction of nitrate. The ammonia was substantially reduced in the free-drained VF bed and nitrate was effectively reduced in the final HF bed. The inflow mean NH₄-N concentration of 29.9 mg/l was reduced to 6.5 mg/l with the average removal efficiency of 78.3%. At the same time the average nitrate-N concentration rose from 0.5 to only 2.7 mg/l at the outflow. Removal of BOD₅ and COD amounted to 94.5% and 84.4%, respectively, with respective average outflow concentrations of 10 and 50 mg/l. Phosphorus was removed efficiently despite the fact that the system was not aimed at P removal and therefore no special media were used. Phosphorus removal amounted in 2008 to 65.4%, but the average outflow concentration of 1.8 mg/l is still high. The results of the present study indicate very efficient performance of the hybrid constructed wetlands, but optimal loading parameters still need to be adjusted. The capital cost of the experimental system is comparable to the conventional on-site treatment plant but the operations and maintenance costs are about one third of the conventional plant.     

Microbially Induced Calcium Carbonate Precipitation on Surface or in the Bulk of Soil

Microbial precipitation of calcium carbonate takes place in nature by different mechanisms. One of them is microbially induced carbonate precipitation (MICP), which is performed due to bacterial hydrolysis of urea in soil in the presence of calcium ions. The MICP process can be adopted to reduce the permeability and/or increase the shear strength of soil. In this paper, a study on the use of Bacillus sp., which was isolated from tropical beach sand, to perform MICP either on the surface or in the bulk of sand is presented. If the level of calcium salt solution was below the sand surface, MICP took place in the bulk of sand. On the other hand, if the level of calcium salt solution was above the sand surface, MICP was performed on the sand surface and formed a thin layer of crust of calcium carbonate. After six sequential batch treatments with suspension of urease-producing bacteria and solutions of urea and calcium salt, the permeability of sand was reduced to 14 mm/day (or 1.6×10^?7 m/s) in both cases of bulk and surface MICP. Quantities of precipitated calcium after six treatments were 0.15 and 0.60 g of Ca per cm² of treated sand surface for the cases of bulk or surface MICP, respectively. The stiffness of the MICP treated sand also increased considerably. The modulus of rupture of the thin layer of crust was 35.9 MPa which is comparable with limestone.     

Municipal wastewater treatment with vertical flow constructed wetlands for irrigation reuse

The treatment effect of two pilot-scale vertical flow constructed wetlands (VFCWs) on municipal wastewaters and their suitability for irrigation reuse were evaluated in a 2-year (2002–2003) experiment. One VFCW was planted with Typha latifolia and the other with Phragmites australis. VFCW efficiency was evaluated in terms of both mass removal and water quality improvement, considering the following parameters: pH, electrical conductivity (EC_w), total suspended solids (TSS), chemical oxygen demand (COD) and biochemical oxygen demand (BOD), total nitrogen (TN) and nitrate (NO₃^?), total phosphorus (TP) and orthophosphate (PO₄^3?), sodium (Na), potassium (K), magnesium (Mg) and calcium (Ca). The accumulation of the elements in the plant organs and VFCW sandy surface layer and their offtake with the macrophyte harvest were also measured.In quantitative terms the established VFCWs showed higher removal efficiencies (>86%) for COD, BOD, N and K, while lower efficiencies (<47%) were observed for Na and Mg. The direct contribution (offtake) of the macrophytes in N, P and K removal processes was particularly high (>65%) due to the massive growth. The results were less favourable in terms of water quality, because the high evapotranspiration losses counteracted the depuration process by concentrating the elements in the outflow water. Higher concentrations were found in outflow than inflow, especially of Na (relative increase of 89%) and Mg (relative increase of 74%). Only parameters with high removal efficiencies fulfilled the Italian guidelines for irrigation reuse whereas parameters with lower efficiencies (e.g., TSS, TP) limited the potential water reuse. Efficient pre-cleaning systems or innovative integrated systems are thus necessary to obtain high removal efficiencies that reduce the effect of ET on water quality.     

Activation of phospholipase C is not correlated to the formation of prostaglandins and superoxide in cultured rat liver macrophages.

This study evaluates the role of inositol phosphates as possible mediators of the activation of phospholipase A2 and NADPH oxidase in cultured rat liver macrophages. Inositol phosphate formation was achieved by zymosan, immune complexes, latex particles and calcium ionophore while the release of arachidonic acid and the formation of prostaglandin E2 was also elicited by phorbol ester and NaF, but not by latex particles; generation of superoxide was obtained by zymosan and phorbol ester only. The kinetics of the formation of inositol phosphates revealed that within the first few minutes after zymosan addition inositol trisphosphate was formed, followed by inositol bisphosphate and inositol monophosphate. Pre-treatment of the cells with dexamethasone or removal of extracellular calcium led to an inhibition of the zymosan-induced formation of inositol phosphates and prostaglandin E2 but had no effect on the generation of superoxide; inhibition of the Na+/H+ exchanger by removal of extracellular sodium ions led to a decrease of the zymosan-induced synthesis of prostaglandin E2, but did not affect the formation of inositol phosphates and superoxide. Pre-treatment of the cells with phorbol ester decreased the zymosan-induced synthesis of prostaglandin E2 and superoxide, but even enhanced the zymosan-induced formation of inositol phosphates. These data indicate that in cultured rat liver macrophages the formation of prostaglandins and superoxide cannot be correlated to an activation of phospholipase C.     

Treatment of domestic wastewater and production of commercial flowers in vertical and horizontal subsurface-flow constructed wetlands

In developing countries, the use of non-conventional plant species as emergent plants in constructed wetlands may add economic benefits besides treating wastewater. In this work, the use of four commercial-valuable ornamental species (Zantedeschia aethiopica, Strelitzia reginae, Anturium andreanum and Agapanthus africanus) was investigated in two types of subsurface wetlands for domestic wastewater treatment. Several water quality parameters were evaluated at the inlet and outlets of a pilot-scale system. Physical measurements were used to evaluate and compare the development of the ornamental plants under two patterns of flow in subsurface wetlands.The results for pollutant removal were significantly higher in the vertical subsurface-flow constructed wetlands (VFCW) for most pollutants. The average removals were more than 80% for BOD and COD; 50.6% for Org-N; 72.2% for NH₄⁺, 50% for Total-P and 96.9% for TC. Only two pollutants were removed in statistically higher percentages in the horizontal subsurface-flow constructed wetlands (HFCW) (NO₃^?, 47.7% and TSS, 82%). The pollutant removal efficiencies were similar to the results obtained in many studies with conventional macrophytes. Most ornamental plants survived the 12-month period of experimentation and their development depended on the type of constructed wetland they were planted. Z. aethiopica looked healthier and produced around 60 flowers in the HFCW. The other three species developed better in the VFCW, although A. andreanum died during the winter. S. reginae produced healthier flowers (and more) and bigger leaves and A. Agapanthus produced more leaves and more lasting flowers. This suggests that it is possible to produce commercial flowers in constructed wetlands without reducing the efficiency of the treatment system.     

Epidermal growth factor stimulates the incorporation of phosphate into phosphatidic acid and phosphoinositides but does not affect phosphoinositide breakdown by phospholipase C in renal cortical slices

The effects of epidermal growth factor (EGF) on the metabolism of phosphatidic acid and phosphoinositides were examined using renal cortical slices labelled with either sodium [³²P]orthophosphate or myo-[³H]inositol. EGF was found to increase the incorporation of phosphate into phosphatidic acid and phosphoinositides. This effect is not dependent on external calcium and is inhibited by 12-O-tetradecanoylphorbol 13-acetate (TPA). When phospholipids were prelabelled, EGF did not decrease the level of ³²P in phosphatidic acid and phosphoinositides, and EGF did not affect the formation of inositol phosphates or the concentration of cAMP and cGMP in renal tissue. The results show that EGF stimulates the incorporation of phosphate into phosphatidic acid and phosphoinositides, but does not affect breakdown of phosphoinositides by phospholipase C in renal cortical slices.     

Bacterial Reductive Dissolution of Crystalline Fe(III) Oxide in Continuous-Flow Column Reactors

Bacterial reductive dissolution of synthetic crystalline Fe(III) oxide-coated sand was studied in continuous-flow column reactors in comparison with parallel batch cultures. The cumulative amount of aqueous Fe(II) exported from the columns over a 6-month incubation period corresponded to (95.0 ± 3.7)% (n = 3) of their original Fe(III) content. Wet-chemical analysis revealed that only (6.5 ± 3.2)% of the initial Fe(III) content remained in the columns at the end of the experiment. The near-quantitative removal of Fe was visibly evidenced by extensive bleaching of color from the sand in the columns. In contrast to the column reactors, Fe(II) production quickly reached an asymptote in batch cultures, and only (13.0 ± 2.2)% (n = 3) of the Fe(III) oxide content was reduced. Sustained bacterial-cell growth occurred in the column reactors, leading to the production and export of a quantity of cells 100-fold greater than that added during inoculation. Indirect estimates of cell growth, based on the quantity of Fe(III) reduced, suggest that only an approximate doubling of initial cell abundance was likely to have occurred in the batch cultures. Our results indicate that removal of biogenic Fe(II) via aqueous-phase transport in the column reactors decreased the passivating influence of surface-bound Fe(II) on oxide reduction activity, thereby allowing a dramatic increase in the extent of Fe(III) oxide reduction and associated bacterial growth. These findings have important implications for understanding the fate of organic and inorganic contaminants whose geochemical behavior is linked to Fe(III) oxide reduction.     

Nitrogen and phosphorus removal from wastewater by subsurface wetlands planted with Iris pseudacorus

Subsurface horizontal flow constructed wetlands are being evaluated for nitrogen (N) and phosphorus (P) removal from wastewater in this study through different gravel sizes, plant densities (Iris pseudacorus), effects of retention times (1 to 10 days) on N and P removal in continuously fed gravel wetland. The inlet and outlet samples were analyzed for TKN, NH₄-N, and NO₃-N, as standard methods. The planted wetland reactor with fine (SG) and coarse (BG) gravels removed 49.4% and 31.4% TKN, respectively, while unplanted reactors removed 43.4% and 26.8% TKN. Also, the efficiencies for NH₄-N were 36.7–43% and 21.6–25.4% for SG and BG planted reactors, respectively. The efficiencies for NO₃-N were 53.5–62.5% and 21.6–25.4% for SG and BG planted reactors, respectively. Roles of plants in SG reactors for O-PO₄ were 5–12% and 3–8% in BG. Also, the roles of plants in the reactors for TP were 9% and 7.4%. The minimum effective detention time for the removal of NO₃-N was 4–5 days. The subsurface constructed wetlands planted with I. pseudacorus can be an appropriate alternative in wastewater treatment natural system in small communities.     

A STUDY ON THE MECHANISM OF INTERCELLULAR ADHESION : Effects of Neuraminidase, Calcium, and Trypsin on the Aggregation of Suspended HeLa Cells

Aggregation of suspended HeLa cells is increased on removal of cell surface sialic acid. Calcium ions promote aggregation whereas magnesium ions have no effect. The calcium effect is abolished by previous treatment of the cells with neuraminidase. Trypsinization of the HeLa cells followed by thorough washing diminishes the rate of mutual cell aggregation. Subsequent incubation with neuraminidase restores the aggregation rate to the original value before trypsin treatment. Cells which had acquired a greater tendency for aggregation after removal of peripheral sialic acid lose this property when subsequently treated with trypsin. Calcium ions have no aggregative effect on trypsinized cells. In contrast to HeLa cells, aggregation of human erythrocytes was not increased after treatment with neuraminidase or on addition of calcium. The results with HeLa cells are interpreted as follows: (a) Trypsin-releasable material confers adhesiveness upon the cells. (b) The adhesive property of this material is counteracted by the presence of cell surface sialic acids. (c) Calcium ions exert their effect by attenuating the adverse effect of sialic acid.     

Estrogen removal from treated municipal effluent in small-scale constructed wetland with different depth

The presence of estrone (E1), 17 beta-estradiol (E2) and 17 alpha-ethynylestradiol (EE2) in sewage treatment work (STW) effluent pose a potential risk to aquatic ecosystem. The objectives of this study were to evaluate the effectiveness of vertical-flow wetland as polishing step of conventional wastewater treatment in the removal of estrogens and to examine the effect of sand depth. The highest removal efficiency of 67.8 ± 28.0%, 84.0 ± 15.4% and 75.3 ± 17.6% for E1, E2 and EE2, respectively, was achieved by the shallowest wetland among three constructed wetlands with different filter layer depth (i.e. 7.5, 30 and 60 cm). Together with the result that the performance of wetlands when operating in unsaturated condition was superior to that when operating in water-saturated condition, it is suggested that maintaining sufficient aerobic circumstance in constructed wetlands was important for estrogens removal. Core sampling indicated that the highest efficiency achieved in extremely shallow wetland might be due partly to the highest root density, besides the superior condition for penetration of oxygen. The adsorbed estrogens in sand accounted for less than 12% of the removed estrogens irrespective of the depth, indicating biotic processes play a major role in the estrogens removal.