The removal of nutrients from non-point source wastewater by a hybrid bioreactor

The aim of this project was to establish an economical and environmentally benign biotechnology for removing nutrients from non-point source wastewater. The proposal involves a hybrid bioreactor comprised of sequential anaerobic, anoxic and aerobic (A²/O) processes and an eco-ditch being constructed and applied in a suburban area, Kunming, south-western China, where wastewater was discharged from an industrial park and suburban communities. The results show that the hybrid bioreactor fosters heterotrophic and autotrophic microorganisms. When the hydraulic load is 200 m³ per day with the running mode in 12 h cycles, the removal efficiencies of the nutrients were 81% for TP, 74% for TDP, 82% for TN, 79% for NO₃-N and 86% for NH₄-N. The improved bacterial community structure and bacterial habitats further implied enhanced water quality and indicates that the easily-deployed, affordable and environmentally-friendly hybrid bioreactor is a promising bio-measure for removing high loadings of nutrients from non-point source wastewater.     

A multi-level bioreactor to remove organic matter and metals, together with its associated bacterial diversity

The purpose of this study was to treat complex wastewater consisting of domestic wastewater, tobacco processing and building materials washings. The proposed multi-level bioreactor consists of a biopond-biofilter, anoxic/aerobic (A/O) fluidized beds and a photoautotrophic system. The results show that when the hydraulic load of the bioreactor was 200m3/d, it successfully and simultaneously removed the organic matter and metals. When the bioreactor was in a relatively steady-state condition, the overall average organic matter and metals removal efficiencies are as follows, COD (89%), UV₂₄₅ nm-matter (91%), Cu (78%), Zn (79%) and Fe (84%). The growth conditions of the native bacterial habitat were improved, which resulted from the increase of the in bacterial diversity under the rejuvenated conditions induced by the bioreactor. The results demonstrate that the multi-level bioreactor, without a sludge treatment system, can remove heterogeneous organic matter and metals from wastewater.     

Analysis of bacteria communities in an up-flow fixed-bed (UFB) bioreactor for treating sulfide in hydrocarbon wastewater

An up-flow fixed-bed (UFB) bioreactor with patented functional polyurethane foam (FPUF) carriers was used to treat sulfide in hydrocarbon wastewater. Community compositions of autotrophic and heterotrophic bacteria were analyzed by polymerase chain reaction–denaturing gradient gel electrophoresis (PCR–DGGE). DGGE results showed that a relatively stable bacterial community composed of heterotrophic and autotrophic bacteria formed in the bioreactor by the end of experiment, which ensured 92–100% sulfide removal efficiencies. Furthermore, autotrophic genera of Thiobacillus and Thiomonas, as well as those of the heterotrophic genus of Acinetobacter survived and exhibited high sulfide oxidation activity under all three operational conditions. Different special genera were also observed under each operational condition, such as the halophilic genus of Nesterenkonia. In addition, a new genus of sulfide oxidation bacteria was found in the bioreactor, which had the ability to synthesize cytoplasm from organic compounds. These genera have wide applications for the treatment of sulfide in hydrocarbon wastewater.     

Accumulation of nickel ion (Ni2+) by immobilized cells ofEnterobacter species

Summary AnEnterobacter species, isolated from electroplating effluent, could remove significant amount of nickel ion (Ni²⁺) from growth medium and sewage effluent. In order to construct a bioreactor to remove Ni²⁺ from electroplating effluent, bacterial cells were immobilized in polyacrylamide beads. The highest removal capacity (RC, mg of Ni²⁺/g of dry cells) and removal efficiency (RE, % of added Ni²⁺ removed by bacterial cells) of Ni²⁺ of immobilized bacterial cells were obtained by optimizing the growth conditions for the bacterial cells such as the composition of the growth medium, incubation time and incubation temperature; and the operational parameters of the bioreactor such as retention time and pH of the Ni²⁺ containing solution, respectively.     

Removal of benzene in a hybrid bioreactor

A novel hybrid bioreactor was designed to remove volatile organic compounds from wastewater and its performance was investigated. The bioreactor was composed of a biofilter section and a bubble column bioreactor section. Benzene was used as a model compound and the influent benzene was removed by immobilized cells in a bubble column bioreactor. Gas phase benzene stripped by air injection was removed in a biofilter. When the superficial air flow rate was 21.1 m h⁻¹ (0.76 min of residence time in a biofilter), up to 2.2 ppm of benzene in gas phase was removed completely in a biofilter and the maximum removal rate was 4.71 mg day⁻¹ cm⁻³. The concentration profile of benzene along the biofilter column was dependent on the superficial air flow rate and the degree of microbial adaptation. Air flow rate and residence time were found to be the most important operation parameters for the hybrid bioreactor. By manipulating these operational parameters, the removal efficiency and capacity of the hybrid bioreactor could be enhanced. The organic load on the hybrid bioreactor could be shared by the biofilter and bubble column bioreactors and the fluctuation of load on the hybrid bioreactor could be absorbed by changing the distribution of benzene between biofilter and bubble column bioreactors. The maximum removal capacity of the hybrid bioreactor in the experimental range was obtained when the biofilter took 50.3% of influent benzene while 100% of removal efficiency was achieved when the biofilter took 72.3% of influent benzene.     

Nitrogen-removal bioreactor capable of simultaneous nitrification and denitrification for application to industrial wastewater treatment

A bioreactor system with 30 packed gel envelopes was installed in a thermal power plant for the removal of nitrogen from ammonia-containing desulfurization wastewater. Each envelope consisted of double-sided plate gels containing Nitrosomonas europaea and Paracoccus denitrificans cells with an internal space in between for injecting an electron donor. The envelope can remove ammonia from wastewater in a single step. When the wastewater was continuously treated with the bioreactor system, it removed 95.0% of the total nitrogen in the inlet, and the total nitrogen concentration in the outlet was below 9.0 mg L⁻¹. The maximum nitrogen removal rate was 6.0 g day⁻¹ per square meter of the gel area. The maximum utilization efficiency of the injected ethanol for denitrification was 98.4%, and the total organic carbon concentration in the outflow was maintained at a low level. Since the bioreactor system could use the electron donor effectively, it was not necessary to use an additional aerobic tank to remove the electron donor and a settling tank to segregate the surplus sludge containing bacteria from wastewater. Our concept of using packed gel envelopes would be highly effective for constructing a simple and efficient nitrogen removal system capable of simultaneous nitrification and denitrification.     

Reactive black-5 azo dye treatment in suspended and attach growth sequencing batch bioreactor using different co-substrates

Treatment of textile wastewater is a big challenge because of diverse chemical composition, high chemical strength and color of the wastewater. In the present study, treatment of wastewater containing reactive black-5 azo dye was studied in anaerobic sequencing batch bioreactor (SBBR) using mixed liquor suspended solids (MLSS) from suspended and attach growth bioreactors. MLSS at concentration of 1000 mg/L and reactive black-5 azo dye at 100 mg/L were used. A culture (10⁸–10⁹ CFU/ml) of pre-isolated bacterial strains (Psychrobacter alimentarius KS23 and Staphylococcus equorum KS26)) capable of degrading azo dyes in mineral salt medium was used to accelerate the treatment process in bioreactor. Different combinations of sludge, culture and dye were used for treatment using different co-substrates. About 85% COD removal was achieved by consortium (MLSS + KS23 + KS26) after 24 h in attach growth bioreactor. Similarly, 92% color removal was observed with consortium in attach growth bioreactor compared to 85% color removal in suspended bioreactor. Addition of bacterial culture (20%, v/v) to the bioreactor could enhance the rate of color removal. This study suggests that biotreatment of wastewater containing textile dyes can be achieved more efficiently in the attach growth bioreactor using yeast extract as a co-substrate and MLSS augmented with dye-degrading bacterial strains.     

Comparison of the Heavy Metal Biosorption Capacity of Active,Heat‐Inactivated and NaOH‐Treated Phanerochaete chrysosporium Biosorbents

Three different kinds of Phanerochaete chrysosporium (NaOH‐treated, heat‐inactivated and active) biosorbent were used for the removal of Cd(II) and Hg(II) ions from aquatic systems. The biosorption of Cd(II) and Hg(II) ions on three different forms of Phanerochaete chrysosporium was studied in aqueous solutions in the concentration range of 50–700 mg/L. Maximum biosorption capacities of NaOH‐treated, heat‐inactivated and active Phanerochaete chrysosporium biomass were found to be 148.37 mg/g, 78.68 mg/g and 68.56 mg/g for Cd(II) as well as 224.67 mg/g, 122.37 mg/g and 88.26 mg/g for Hg(II), respectively. For Cd(II) and Hg(II) ions, the order of affinity of the biosorbents was arranged as NaOH‐treated > heat‐inactivated > active. The order of the amount of metal ions adsorbed was established as Hg(II) > Cd(II) on a weight basis, and as Cd(II) > Hg(II) on a molar basis. Biosorption equilibriums were established in about 60 min. The effect of the pH was also investigated, and maximum rates of biosorption of metal ions on the three different forms of Phanerochaete chrysosporium were observed at pH 6.0. The reusability experiments and synthetic wastewater studies were carried out with the most effective form, i.e., the NaOH‐treated Phanerochaete chrysosporium biomass. It was observed that the biosorbent could be regenerated using 10 mM HCl solution, with a recovery of up to 98%, and it could be reused in five biosorption‐desorption cycles without any considerable loss in biosorption capacity. The alkali‐treated Phanerochaete chrysosporium removed 73% of Cd(II) and 81% of Hg(II) ions from synthetic wastewater.     

Comparison of bacterial diversity in full scale anammox bioreactors operated under different conditions

Bacterial community structure of full‐scale anammox bioreactor is still mainly unknown. It has never been analyzed whether different anammox bioreactor configurations might result in the development of different bacterial community structures among these systems. In this work, the bacterial community structure of six full‐scale autotrophic nitrogen removal bioreactors located in The Netherlands and China operating under three different technologies and with different influent wastewater characteristics was studied by the means of pyrotag sequencing evaluation of the bacterial assemblage yielded a great diversity in all systems. The most represented phyla were the Bacteroidetes and the Proteobacteria, followed by the Planctomycetes. 14 OTUs were shared by all bioreactors, but none of them belonged to the Brocadiales order. Statistical analysis at OTU level showed that differences in the microbial communities were high, and that the main driver of the bacterial assemblage composition was different for the distinct phyla identified in the six bioreactors, depending on bioreactor technology or influent wastewater characteristics. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:1464–1472, 2015     

Experimental and theoretical approaches for Cd(II) biosorption from aqueous solution using Oryza sativa biomass

Biomass of Oryza sativa (OS) was tested for the removal of Cd(II) ions from synthetic and real wastewater samples. Batch experiments were conducted to investigate the effects of operating parameters on Cd(II) biosorption. Fourier transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive x-ray spectroscopy were used to examine the surface characteristics of the Cd(II)-loaded biomass. The maximum removal efficiency of Cd(II) was 89.4% at optimum pH 6.0, biosorbent dose 10.0 g L^?1, initial Cd(II) 50 mg L^?1, and biosorbent particle size 0.5 mm. The applicability of Langmuir and Freundlich isotherms to the sorbent system implied the existence of both monolayer and heterogeneous surface conditions. Kinetic studies revealed that the adsorption process of Cd(II) followed the pseudo-second-order model (r2: 0.99). On the theoretical side, an adaptive neuro-fuzzy inference system (ANFIS) was applied to select the operating parameter that mostly influences the Cd(II) biosorption process. Results from ANFIS indicated that pH was the most influential parameter affecting Cd(II) removal efficiency, indicating that the biomass of OS was strongly pH sensitive. Finally, the biomass was confirmed to adsorb Cd(II) from real wastewater samples with removal efficiency close to 100%. However, feasibility studies of such systems on a large-scale application remain to be investigated.     

Evaluation of Lupinus Species to Accumulate Heavy Metals From W aste Waters

Several Lupinus species, for example, Lupinus albus, Lupinus luteus, Lupinus angustifolius, and Lupinus hispanicus were used to accumulate Mn(II), Cd(II), Pb(II), Cr(III), Cr(VI), Hg²⁺, and CH₃Hg⁺ from waste waters. The influence of different species concentrations (50 and 100 mg L^-1) and pH on growing behavior as well as the resulting distribution of metals in the plants were investigated. The results obtained showed that lupins were able to germinate and to grow in the presence of the metals mentioned above, even when they were present at levels as high as 50 mg L^-1. Accumulation of Pb(II), Cr(III), and Cd(II) was higher in roots than in shoots. As far as mercury is concerned, the highest CH₃Hg and Hg²⁺ accumulation was detected in roots, but fast transport toward the leaves was noticed. In contrast to mercury, the uptake of chromium seems to be influenced by the chemical form of the analyte, remaining Cr(VI) in solution. No differences in growing behavior and accumulation were observed for the four Lupinus species studied. Even though plants were exposed only a relatively short time to the metal solutions, metal concentrations of approximately 2 g/kg of dry matter were detected in the young lupins plants. The feasibility of utilizing Lupinus plants for the removal of heavy metals from wastewater was also investigated. Lupins were able to grow under extreme conditions (wastewater, pH lower than 2) and to remove 98% of the initial amount of toxic metals present in the sample.     

Nitrifying bacterial growth inhibition in the presence of algae and cyanobacteria

Nitrifying bacteria, cyanobacteria, and algae are important microorganisms in open pond wastewater treatment systems. Nitrification involving the sequential oxidation of ammonia to nitrite and nitrate, mainly due to autotrophic nitrifying bacteria, is essential to biological nitrogen removal in wastewater and global nitrogen cycling. A continuous flow autotrophic bioreactor was initially designed for nitrifying bacterial growth only. In the presence of cyanobacteria and algae, we monitored both the microbial activity by measuring specific oxygen production rate (SOPR) for microalgae and cyanobacteria and specific oxygen uptake rate (SOUR) for nitrifying bacteria. The growth of cyanobacteria and algae inhibited the maximum nitrification rate by a factor of 4 although the ammonium nitrogen fed to the reactor was almost completely removed. Terminal restriction fragment length polymorphism (T‐RFLP) analysis indicated that the community structures of nitrifying bacteria remained unchanged, containing the dominant Nitrosospira, Nitrospira, and Nitrobacter species. PCR amplification coupled with cloning and sequencing analysis resulted in identifying Chlorella emersonii and an uncultured cyanobacterium as the dominant species in the autotrophic bioreactor. Notwithstanding their fast growth rate and their toxicity to nitrifiers, microalgae and cyanobacteria were more easily lost in effluent than nitrifying bacteria because of their poor settling characteristics. The microorganisms were able to grow together in the bioreactor with constant individual biomass fractions because of the uncoupled solids retention times for algae/cyanobacteria and nitrifiers. The results indicate that compared to conventional wastewater treatment systems, longer solids retention times (e.g., by a factor of 4) should be considered in phototrophic bioreactors for complete nitrification and nitrogen removal. Biotechnol. Bioeng. 2010;107: 1004–1011. © 2010 Wiley Periodicals, Inc.     

Phosphate uptake and release by Acinetobacter johnsonii in continuous culture and coupling of phosphate release to heavy metal accumulation

A strain of polyphosphate-synthesizing, phosphate-releasing Acinetobacter johnsonii was isolated from a wastewater treatment plant operating enhanced biological phosphate removal (EBPR) and was used to remove La³⁺ from solution via precipitation of cell-bound LaPO₄. The effect of repeated aerobic–anaerobic cycles on the carbon and phosphate metabolism of the organism was studied in attempts to promote increased phosphate flux using a three-stage, continuous bioreactor comprising aerobic, anaerobic and settling vessels. The bioreactor was operated in two modes: In flow-through mode, cells were grown aerobically with acetate as the sole carbon source, promoting excess phosphate uptake (up to 5.0 mmol/l=3.0 mmol/g protein). Cells were diluted into the anaerobic vessel where phosphate was released (up to 1.0 mmol/l=0.3 mmol/g protein), and thence to waste. The system was initially operated to steady state in flow-through mode, then switched to recycle mode. Here the anaerobic vessel output passed to a settling vessel from which settled cells were returned to the aerobic vessel. Carbon source (acetate) was supplied only to the anaerobic vessel; increased anaerobic acetate uptake was observed during recycle, which was sustained when the system was returned to flow-through mode and was related to increased cellular lipid inclusions by flow cytometry and electron microscopy. These phenomena may represent adaptation of cells to aerobic–anaerobic cycling with aerobic carbon/energy limitation. Addition of La³⁺ to the anaerobic vessel during recycle mode promoted removal of 95% of the La³⁺ from a 0.1 to 0.3 mM (14–42 ppm) solution at the expense of biogenic phosphate. Journal of Industrial Microbiology & Biotechnology (2001) 26, 333–340. Received 05 July 2000/ Accepted in revised form 12 April 2001     

Carbon source recovery from waste activated sludge by alkaline hydrolysis and gamma-ray irradiation for biological denitrification

The recovery of an organic carbon source from a waste activated sludge by using alkaline hydrolysis and radiation treatment was studied, and the feasibility of the solubilized sludge carbon source for a biological denitrification was also investigated. The effects of an alkaline treatment and gamma-ray irradiation on a biodegradability enhancement of the sludge were also studied. A modified continuous bioreactor for a denitrification (MLE reactor) was operated by using a synthetic wastewater for 47 days. Alkaline treatment of pH 10 and gamma-ray irradiation of 20 kGy were found to be the optimum carbon source recovery conditions. COD removal of 84% and T-N removal of 51% could be obtained by using the solubilized sludge carbon source through the MLE denitrification process. It can be concluded that the carbon source recovered from the waste activated sludge was successfully employed as an alternative carbon source for a biological denitrification.     

Exploring the Cadmium Tolerance and Removal Capability of a Filamentous Fungus Fusarium solani

Abstract

Cadmium is one of the most toxic contaminant causing many problems to human health and the environment. These days the world is moving toward ecofriendly and efficient techniques to remove the pollutant from the wastewater. The present study aims to investigate the tolerance of Fusarium solani toward cadmium (Cd), nickel (Ni), and lead (Pb). Maximum tolerance was observed with Cd. Cadmium removal ability of F. solani was examined from contaminated PDB medium. pH, initial concentration and time optimization for maximum removal of Cd by F. solani was also studied. The maximum removal (92.4%) was recorded at initial concentration of 50?mg/L after 144?h of incubation. Cadmium exposure increased the level of glutathione (GSH) and oxidized glutathione (GSSG) contents and the activity of catalase (CAT) in F. solani. Fourier-transform infrared spectroscopy (FTIR) analysis indicated the involvement of the different surface functional group in biosorption of Cd while Scanning Electron Microscopy coupled with Energy Dispersive Spectroscopy (SEM-EDS) analysis revealed the presence of Cd on the surface of fungal cell. The changes observed in compositions of S, P, and Cd using EDS analysis on biomass surface indicated the precipitation of Cd as CdS and Cd₃(PO₄)₂. The XRD analysis revealed the presence of Cd₃(PO₄)₂ on mycelia surface of F. solani.     

Microbial community analysis of a full-scale DEMON bioreactor

Full-scale applications of autotrophic nitrogen removal technologies for the treatment of digested sludge liquor have proliferated during the last decade. Among these technologies, the aerobic/anoxic deammonification process (DEMON) is one of the major applied processes. This technology achieves nitrogen removal from wastewater through anammox metabolism inside a single bioreactor due to alternating cycles of aeration. To date, microbial community composition of full-scale DEMON bioreactors have never been reported. In this study, bacterial community structure of a full-scale DEMON bioreactor located at the Apeldoorn wastewater treatment plant was analyzed using pyrosequencing. This technique provided a higher-resolution study of the bacterial assemblage of the system compared to other techniques used in lab-scale DEMON bioreactors. Results showed that the DEMON bioreactor was a complex ecosystem where ammonium oxidizing bacteria, anammox bacteria and many other bacterial phylotypes coexist. The potential ecological role of all phylotypes found was discussed. Thus, metagenomic analysis through pyrosequencing offered new perspectives over the functioning of the DEMON bioreactor by exhaustive identification of microorganisms, which play a key role in the performance of bioreactors. In this way, pyrosequencing has been proven as a helpful tool for the in-depth investigation of the functioning of bioreactors at microbiological scale.     

Pyrosequence analysis of bacterial communities in aerobic bioreactors treating polycyclic aromatic hydrocarbon-contaminated soil

Two aerobic, lab-scale, slurry-phase bioreactors were used to examine the biodegradation of polycyclic aromatic hydrocarbons (PAHs) in contaminated soil and the associated bacterial communities. The two bioreactors were operated under semi-continuous (draw-and-fill) conditions at a residence time of 35 days, but one was fed weekly and the other monthly. Most of the quantified PAHs, including high-molecular-weight compounds, were removed to a greater extent in the weekly-fed bioreactor, which achieved total PAH removal of 76%. Molecular analyses, including pyrosequencing of 16S rRNA genes, revealed significant shifts in the soil bacterial communities after introduction to the bioreactors and differences in the abundance and types of bacteria in each of the bioreactors. The weekly-fed bioreactor displayed a more stable bacterial community with gradual changes over time, whereas the monthly-fed bioreactor community was less consistent and may have been more strongly influenced by the influx of untreated soil during feeding. Phylogenetic groups containing known PAH-degrading bacteria previously identified through stable-isotope probing of the untreated soil were differentially affected by bioreactor conditions. Sequences from members of the Acidovorax and Sphingomonas genera, as well as the uncultivated “Pyrene Group 2” were abundant in the bioreactors. However, the relative abundances of sequences from the Pseudomonas, Sphingobium, and Pseudoxanthomonas genera, as well as from a group of unclassified anthracene degraders, were much lower in the bioreactors compared to the untreated soil.     

Treatment of high strength organic wastewater by a mixed culture of photosynthetic microorganisms

The growth characteristics and nutrient removal fromsynthetic wastewater by Rhodobacter sphaeroides,Chlorella sorokiniana and Spirulinaplatensis were investigated under aerobic dark(heterotrophic) and aerobic light (photoheterotrophic)conditions. Both in terms of economy and efficiency,aerobic dark conditions were the best for wastewatertreatment using R. sphaeroides and C.sorokiniana, but light was necessary with S.platensis. Neither growth nor nutrient removalcharacteristics of the cells were affected insynthetic wastewater with as high as 10 000 ppmacetate, 1000 ppm propionate, 700 ppm nitrate and 100 ppmphosphate. Although R. sphaeroides and C. sorokiniana showed good growth in syntheticwastewater containing 400 ppm of ammonia, S.platensis was completely inhibited.When grown as a monoculture, none of thestrains could simultaneously remove acetate,propionate, ammonia, nitrate and phosphate from thewastewater. R. sphaeroides could remove allthe above nutrients except nitrate, but the rate of removal was relatively low. The rate of nutrientsremoval by C. sorokiniana was higher, but theorganism could not remove propionate; S.platensis could efficiently remove nitrate, ammoniaand phosphate, but none of the organic acids. A mixedculture of R. sphaeroides and C.sorokiniana was therefore used for simultaneousremoval of organic acids, nitrate, ammonia andphosphate. The optimum ratio of the cells depended onthe composition of the wastewater.     

Biodegradation of sulfamethoxazole by individual and mixed bacteria

Antibiotic compounds, like sulfamethoxazole (SMX), have become a concern in the aquatic environment due to the potential development of antibacterial resistances. Due to excretion and disposal, SMX has been frequently detected in wastewaters and surface waters. SMX removal in conventional wastewater treatment plants (WWTPs) ranges from 0% to 90%, and there are opposing results regarding its biodegradability at lab scale. The objective of this research was to determine the ability of pure cultures of individual and mixed consortia of bacteria (Bacillus subtilis, Pseudomonas aeruginosa, Pseudomonas putida, Rhodococcus equi, Rhodococcus erythropolis, Rhodococcus rhodocrous, and Rhodococcus zopfii) known to exist in WWTP activated sludge to remove SMX. Results showed that R. equi alone had the greatest ability to remove SMX leading to 29% removal (with glucose) and the formation of a metabolite. Degradation pathways and metabolite structures have been proposed based on the potential enzymes produced by R. equi. When R. equi was mixed with other microorganisms, a positive synergistic effect was not observed and the maximum SMX removal achieved was 5%. This indicates that pure culture results cannot be extrapolated to mixed culture conditions, and the methodology developed here to study the biodegradability of compounds under controlled mixed culture conditions offers an alternative to conventional studies using pure bacterial cultures or inocula from activated sludge sources consisting of unknown and variable microbial populations.     

Occurrence of Pathogenic Free‐Living Amoebae and Bacterial Indicators in a Constructed Wetland Treating Domestic Wastewater from a Single Household

Constructed wetlands are effective wastewater treatment systems because of their ability to remove large amounts of organic matter and pathogens. The goals of this study were to characterize the presence of pathogenic free‐living amoebae and bacterial indicators (total and fecal coliforms), and to ascertain the removal efficiencies of physical and chemical pollutants, in a constructed wetland treating domestic wastewater from a single household. Influent and effluent samples were collected monthly over a ten‐month period for biological, physical and chemical analyses. Thirty‐two species of free‐living amoebae were isolated from the system. The genus Acanthamoeba was the most frequently encountered (59 %) and was removed from the wastewater with the greatest efficiency (80 %). Removal of bacteria was low, the highest removal rates were found in August (4 logarithmic units) and January (3 logarithmic units). The average removal efficiencies of suspended solids, BOD₅ and ammoniacal nitrogen were 71.5 %, 50.6 % and 13.1 %, respectively. The relatively low removal efficiencies of the various bacteriological, physical and chemical parameters suggest that the hydraulic retention time was probably insufficient for optimal treatment to occur. The effluent quality was unacceptable for unrestricted irrigation of crops that are eaten uncooked.