Reduction of indicator and pathogenic microorganisms by psychrophilic anaerobic digestion in swine slurries

The objective of this study was to evaluate the efficiency of a low temperature anaerobic treatment to reduce viable populations of indicator microorganisms (total coliforms, Escherichia coli) and the presence of selected pathogens (Salmonella, Yersinia enterocolitica, Cryptosporidium and Giardia) in swine slurries from different sources. Experiments were carried out in 40 l Sequencing Batch Reactors (SBRs). Experimental results indicated that anaerobic digestion of swine manure slurry at 20 degrees C for 20 days in an intermittently fed SBR: (1) reduced indigenous populations of total coliforms by 97.94-100%; (2) reduced indigenous populations of E. coli by 99.67-100%; (3) resulted in undetectable levels of indigenous strains of Salmonella, Cryptosporidium, and Giardia. It can be considered as a promising method for reducing indigenous indicator and pathogenic microorganisms populations in liquid swine manure slurries.     

Challenges and innovations on biological treatment of livestock effluents

Intensification of animal production led to high amounts of manure to be managed. Biological processes can contribute to a sustainable manure management. This paper presents the biological treatments available for the treatment of animal manure, mainly focusing on swine manure, including aerobic processes (nitrification, denitrification, enhanced biological phosphorus removal) and anaerobic digestion. These processes are discussed in terms of pollution removal, ammonia and greenhouse gas emissions (methane and nitrous oxide) and pathogen removal. Application of emerging processes such as partial nitrification and anaerobic ammonium oxidation (anammox) applied to animal manure is also considered. Finally, perspectives and future challenges for the research concerning biological treatments are highlighted in this paper.     

Coliphages and indicator bacteria in birds around Boston Harbor

Droppings from feral populations of pigeons, geese and herring gulls from the urban/suburban environment around Boston Harbor, MA, USA contained up to 10⁶ somatic coliphages, 10⁸ enterococci, 10⁹ thermotolerant coliforms and 10² F-specific coliphages per gram of feces. Somatic coliphages, enterococci and thermotolerant coliforms were common in the feces of all three kinds of birds but F-specific coliphages were found in droppings from only three of 32 gulls. Thus these sources of bacterial and viral indicators should be considered when dealing with the ecology of fecal pollution indicators. Moreover, microbial indicators of fecal or sewage pollution originating from bird droppings may be mistaken for indicators that come from humans. This may cause an overestimate of the hazard from human pathogens in water and confound attempts to locate sources of fecal or sewage pollution. Received 12 May 1998/ Accepted in revised form 15 July 1998     

Comparison of composting, storage and urea treatment for sanitising of faecal matter and manure

To close the loop of nutrients in a safe way, the loop of pathogens has to be broken. By sanitising organic fertilisers derived from faeces and manure, the first step in the disease transmission chain is broken. Two alternatives for this treatment, thermophilic composting and ammonia-based treatment, were evaluated and compared to storage treatment. Thermal composting of faecal matter and food waste resulted in a treatment temperature of over 65 degrees C in a 90 L reactor. By using insulation and turning the compost three times during the high temperature period, it is possible to ensure a 5 log(10) reduction of pathogens. Small scale composting of the same material indicated less efficient reduction of faecal bacteria at temperatures around 50 degrees C. In the chemical treatment tested, an addition of 3% N-NH(3) increased the pH to above 9 within 1h and resulted in a good reduction in the indicator organisms for bacteria (Salmonella spp. and faecal coliforms D(r)<0.7 days, Enterococcus spp. D(r)<3 days). Lower addition rates resulted in a longer treatment period needed. The storage period tested resulted in slow reduction of faecal coliforms, Salmonella spp. and Enterococcus spp.     

Development of a second-generation environmentally superior technology for treatment of swine manure in the USA

New swine waste management systems in North Carolina need to meet high performance standards of an environmentally superior technology (EST) regarding nitrogen, phosphorus, heavy metals, pathogens, ammonia and odor emissions, and remain affordable and simple to operate. The objective of this study was to develop a second-generation treatment system that can achieve high EST standards at reduced costs. The system used solids separation, nitrification/denitrification and phosphorus removal/disinfection, and was demonstrated at full-scale on a 5145-head swine farm during three production cycles (15-months). Removal efficiencies were: 98% suspended solids, 97% ammonia, 95% phosphorus, 99% copper and zinc, 99.9% odors, and 99.99% pathogens. The system met EST standards at 1/3 the cost of the previous version. Animal health and productivity were enhanced; hog sales increased 32,900 kg/cycle (5.6%). These results demonstrated that: (1) significant cost reductions were achieved by on-farm implementation and continued engineering improvements, and (2) the new waste management system substantially benefited livestock productivity.     

Occurrence and persistence of bacterial pathogens and indicator organisms in beach sand along the California coast

This report documents the presence of fecal indicators and bacterial pathogens in sand at 53 California marine beaches using both culture-dependent and -independent (PCR and quantitative PCR [QPCR]) methods. Fecal indicator bacteria were widespread in California beach sand, with Escherichia coli and enterococci detected at 68% and 94% of the beaches surveyed, respectively. Somatic coliphages and a Bacteroidales human-specific fecal marker were detected at 43% and 13% of the beaches, respectively. Dry sand samples from almost 30% of the beaches contained at least one of the following pathogens: Salmonella spp., Campylobacter spp., Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus (MRSA), which were detected at 15%, 13%, 14%, and 3% of tested beaches, respectively. Fecal indicators and pathogens were poorly correlated to one another and to land cover. Sands were dry at the time of collection, and those with relatively high moisture tended to have higher concentrations or a more frequent occurrence of both indicators and pathogens. Using culture-dependent assays, fecal indicators decayed faster than pathogens in microcosm experiments using unaltered beach sand seeded with sewage and assessed by culture-dependent assays. The following order of persistence was observed (listed from most to least persistent): Campylobacter > Salmonella > somatic coliphages > enterococci > E. coli > F(+) phages. In contrast, pathogens decayed faster than fecal indicators in culture-independent assays: enterococci > Bacteroidales human-specific marker > Salmonella > Campylobacter. Microcosm experiments demonstrated that both indicators and pathogens were mobilized by wetting with seawater. Decay rates measured by QPCR were lower than those measured with culture-dependent methods. Enterococcal persistence and possible growth were observed for wetted microcosms relative to unwetted controls.     

Validity of the indicator organism paradigm for pathogen reduction in reclaimed water and public health protection

The validity of using indicator organisms (total and fecal coliforms, enterococci, Clostridium perfringens, and F-specific coliphages) to predict the presence or absence of pathogens (infectious enteric viruses, Cryptosporidium, and Giardia) was tested at six wastewater reclamation facilities. Multiple samplings conducted at each facility over a 1-year period. Larger sample volumes for indicators (0.2 to 0.4 liters) and pathogens (30 to 100 liters) resulted in more sensitive detection limits than are typical of routine monitoring. Microorganisms were detected in disinfected effluent samples at the following frequencies: total coliforms, 63%; fecal coliforms, 27%; enterococci, 27%; C. perfringens, 61%; F-specific coliphages, approximately 40%; and enteric viruses, 31%. Cryptosporidium oocysts and Giardia cysts were detected in 70% and 80%, respectively, of reclaimed water samples. Viable Cryptosporidium, based on cell culture infectivity assays, was detected in 20% of the reclaimed water samples. No strong correlation was found for any indicator-pathogen combination. When data for all indicators were tested using discriminant analysis, the presence/absence patterns for Giardia cysts, Cryptosporidium oocysts, infectious Cryptosporidium, and infectious enteric viruses were predicted for over 71% of disinfected effluents. The failure of measurements of single indicator organism to correlate with pathogens suggests that public health is not adequately protected by simple monitoring schemes based on detection of a single indicator, particularly at the detection limits routinely employed. Monitoring a suite of indicator organisms in reclaimed effluent is more likely to be predictive of the presence of certain pathogens, and a need for additional pathogen monitoring in reclaimed water in order to protect public health is suggested by this study.     

Survival of faecal indicator bacteria in bovine manure incorporated into soil

AIMS: Survival of Escherichia coli and enterococci was evaluated in bovine manure incorporated into two Wisconsin soils. METHODS AND RESULTS: Silty clay loam (SCL) and loamy sand (LS) were mixed with fresh bovine manure, exposed daily to 10 h at 22 degrees C/14 h at 9 degrees C, and watered weekly for 12 weeks. Escherichia coli numbers increased 1-2 log cfu g(-1), then decreased < 1 and about 2 log cfu g(-1) in SCL and LS, respectively. Enterococci numbers rose less and then declined faster than those of E. coli. Watering intervals of 3, 7 and 14 days were evaluated in weeks 13-19, but did not affect the slow decline in numbers of E. coli or enterococci. CONCLUSION: Escherichia coli and enterococci may survive at least 19 weeks at 9-21 degrees C in bovine manure/soil, with E. coli surviving better. SIGNIFICANCE AND IMPACT OF THE STUDY: Quantification of E. coli or enterococci in late spring/early summer soil may be useful in indicating recent application of bovine manure.     

Development of environmentally superior technologies in the US and policy

“Environmentally superior technology” (EST) represents a research initiative in North Carolina (NC) to develop alternatives to lagoon treatment and land application of swine manure. EST performance standards included impacts of animal waste to surface and groundwater, emission of ammonia and odor, release of disease-transmitting pathogens, and heavy metal contamination of soil and groundwater. Five technologies were shown to meet these standards: a solids separation/nitrification–denitrification/soluble phosphorus removal system; a thermophilic anaerobic digester system; a centralized composting system; a gasification system; and a fluidized bed combustion system. Economic data compiled for all EST showed annualized (10-year) costs of retrofitting existing swine farms with the technologies ranged between $90 and over $400 per 1000 lbs. steady state live-weight. Value-engineering to reduce the costs of targeted EST has been successful. Policy providing institutional incentives to incorporate EST has been enacted as a result of this study.     

Sunlight inactivation of fecal bacteriophages and bacteria in sewage-polluted seawater.

Sunlight inactivation rates of somatic coliphages, F-specific RNA bacteriophages (F-RNA phages), and fecal coliforms were compared in seven summer and three winter survival experiments. Experiments were conducted outdoors, using 300-liter 2% (vol/vol) sewage-seawater mixtures held in open-top chambers. Dark inactivation rates (k(D)s), measured from exponential survival curves in enclosed (control) chambers, were higher in summer (temperature range: 14 to 20 degrees C) than in winter (temperature range: 8 to 10 degrees C). Winter k(D)s were highest for fecal coliforms and lowest for F-RNA phages but were the same or similar for all three indicators in summer. Sunlight inactivation rates (k(S)), as a function of cumulative global solar radiation (insolation), were all higher than the k(D)s with a consistent k(S) ranking (from greatest to least) as follows: fecal coliforms, F-RNA phages, and somatic coliphages. Phage inactivation was exponential, but bacterial curves typically exhibited a shoulder. Phages from raw sewage exhibited k(S)s similar to those from waste stabilization pond effluent, but raw sewage fecal coliforms were inactivated faster than pond effluent fecal coliforms. In an experiment which included F-DNA phages and Bacteroides fragilis phages, the k(S) ranking (from greatest to least) was as follows: fecal coliforms, F-RNA phages, B. fragilis phages, F-DNA phages, and somatic coliphages. In a 2-day experiment which included enterococci, the initial concentration ranking (from greatest to least: fecal coliforms, enterococci, F-RNA phages, and somatic coliphages) was reversed during sunlight exposure, with only the phages remaining detectable by the end of day 2. Inactivation rates under different optical filters decreased with the increase in spectral cutoff wavelength (50% light transmission) and indicated that F-RNA phages and fecal coliforms are more susceptible than somatic coliphages to longer solar wavelengths, which predominate in seawater. The consistently superior survival of somatic coliphages in our experiments suggests that they warrant further consideration as fecal, and possibly viral, indicators in marine waters.     

Characteristics of solids, BOD5 and VFAs in liquid swine manure treated by short-term low-intensity aeration for long-term storage

A laboratory-scale experiment presents data that reveal the temporal characteristics of solids, biochemical oxygen demand (BOD5) and volatile fatty acids (VFAs) in the aerated liquid swine manure for minimizing odor generation potential during 190-day storage. The performance of 15-day aeration of liquid manure with initial total solids (TS) content from 0.5% to 4.0% was examined at low-intensity aeration rates, i.e., +35 mV oxidation-reduction potential (ORP), 1.0 mg O2/l and 3.0 mg O2/l dissolved oxygen (DO). Odor generation potential was evaluated using VFAs. The aeration process contributed remarkably to the decomposition of TS, total volatile solids (TVS), BOD5 and VFAs. Moreover, the stabilization of manure due to aeration could last up to 190 days. The TS reduction on day 190 ranging from 6.3% to 32.7%, 20.2% to 39.1%, 19.0% to 41.0% were realized under the intensities of +35 mV ORP, 1.0 and 3.0 mg O2/l, respectively. At the same time, the reduction of BOD5 and VFAs reached around 7.8% to 69.5%, 17.2% to 79.9% and 21.9% to 91.1%; 0.4% to 91.0%, 60.4% to 95.0% and 70.4% to 94.1%. The liquid manure with low solids (e.g., TS of 0.5% and 1.0%) offered an advantageous condition for aeration treatment, particularly for biodegradation of BOD5 and VFAs. The odor generation potential could also be evaluated by the levels of solids and BOD5 in the manure. Increasing aeration intensity would significantly diminish the odor generation potential for given levels of solids and/or BOD5. Fifteen-day aeration with intensity of 1.0 mg O2/l may be recommended at farm level for both odor control and energy savings.     

Potential Regrowth and Recolonization of Salmonellae and Indicators in Biosolids and Biosolid-Amended Soil

This study evaluated the potential for conversion of Class B to Class A biosolids with respect to salmonellae and fecal coliforms during solar drying in concrete lined drying beds. Anaerobically (8% solids) and aerobically (2% solids) digested Class B biosolids were pumped into field-scale drying beds, and microbial populations and environmental conditions were monitored. Numbers of fecal coliforms and salmonellae decreased as temperature and rate of desiccation increased. After 3 to 4 weeks, Class A requirements were achieved in both biosolids for the pathogens and the indicators. However, following rainfall events, significant increase in numbers was observed for both fecal coliforms and salmonellae. In laboratory studies, regrowth of fecal coliforms was observed in both biosolids and biosolid-amended soil, but the regrowth of salmonellae observed in the concrete-lined drying beds did not occur. These laboratory studies demonstrated that pathogens decreased in numbers when soil was amended with biosolids. Based on serotyping, the increased numbers of salmonellae seen in the concrete lined drying beds following rainfall events was most likely due to recolonization due to contamination from fecal matter introduced by animals and not from regrowth of salmonellae indigenous to biosolids. Overall, we conclude that the use of concrete-lined beds created a situation in which moisture added as rainfall accumulated in the beds, promoting the growth of fecal coliforms and salmonellae added from external sources.     

Potential regrowth and recolonization of salmonellae and indicators in biosolids and biosolid-amended soil

This study evaluated the potential for conversion of Class B to Class A biosolids with respect to salmonellae and fecal coliforms during solar drying in concrete lined drying beds. Anaerobically (8% solids) and aerobically (2% solids) digested Class B biosolids were pumped into field-scale drying beds, and microbial populations and environmental conditions were monitored. Numbers of fecal coliforms and salmonellae decreased as temperature and rate of desiccation increased. After 3 to 4 weeks, Class A requirements were achieved in both biosolids for the pathogens and the indicators. However, following rainfall events, significant increase in numbers was observed for both fecal coliforms and salmonellae. In laboratory studies, regrowth of fecal coliforms was observed in both biosolids and biosolid-amended soil, but the regrowth of salmonellae observed in the concrete-lined drying beds did not occur. These laboratory studies demonstrated that pathogens decreased in numbers when soil was amended with biosolids. Based on serotyping, the increased numbers of salmonellae seen in the concrete lined drying beds following rainfall events was most likely due to recolonization due to contamination from fecal matter introduced by animals and not from regrowth of salmonellae indigenous to biosolids. Overall, we conclude that the use of concrete-lined beds created a situation in which moisture added as rainfall accumulated in the beds, promoting the growth of fecal coliforms and salmonellae added from external sources.     

Evaluating the prevalence and genetic diversity of adenovirus and polyomavirus in bovine waste for microbial source tracking

This study evaluated and compared the occurrence, concentrations, and genetic diversity of bovine polyomavirus (BPyV) and bovine adenovirus (BAdV) in manure and feces samples for the purpose of determining which of these two viruses is more suitable for bovine fecal indication and microbial source tracking. The comparability and correlation between concentrations/prevalence of these viruses and bacterial fecal indicators (cow-associated Bacteroidetes, Escherichia coli and enterococci) in manure and fecal samples was also determined. A total of 26 dairy manure and 18 individual dairy cow feces samples were tested. The results showed the mean concentration of BAdV in all of dairy manure samples was at least 1 log lower than BPyV (p ≤ 0.005). All of the dairy manure samples tested positive for BPyV but not for BAdV. After combining dairy manure measurements, bacterial indicators had 0.3–0.7 log (p ≤ 0.05) and 1.8–2.2 log (p ≤ 0.005) higher concentrations than BPyV and BAdV, respectively. The concentration of BPyV had a significant positive correlation with the concentration of E. coli and enterococci in the manure samples. The partial VP1 genetic sequences of BPyV isolated from three different farms had a 100% homology to each other and to VP1 sequence (D13942.1) reported in previous study. Based on the occurrence, quantitative and genetic diversity results, BPyV may be a better indicator than BAdV for microbial source tracking at manure application sites.     

Effects of storage on characteristics and hygienic quality of digestates from four co-digestion concepts of manure and biowaste

This study evaluated the effects of storage in northern winter conditions (5 degrees C) on the characteristics and nutrients separation of digestates from co-digestion of manure and biowaste as well as the hygienic quality of the digestates after digestion and storage. During 3-11 months' storage average nitrogen losses and reductions of total solids (TS) and volatile solids (VS) were 0-15%. With some exceptions, soluble chemical oxygen demand (SCOD) had increased slightly (from approximately 6.5 to approximately 7.5g/l) after 3 months' storage, while after 9-11 months' it had decreased from 8.3-11 to 5.6-8.4g/l. The concentrations of P(tot) and PO4-P in the separated liquid fractions decreased 40-57% after 3 months' storage and 71-91% after 9 months' storage compared to the initial concentrations. The methane potential losses during 9-11 months' storage corresponded 0-10% of the total methane potential without storage. The hygienic quality of the digestates from the 55 degrees C reactor and during storage fulfilled the Animal By-Products Regulation (ABPR) demands while the 35 degrees C digestate contained 0-105cfu/g of indicator bacteria (faecal coliforms, enterobacteria, enterococcus) and >10cfu/g of spiked salmonella, which amounts decreased slowly during storage. Sulphite reducing clostridia was not affected by either digestion or storage.     

Automated electrical impedance technique for rapid enumeration of fecal coliforms in effluents from sewage treatment plants.

Fecal coliforms growing in a selective lactose-based broth medium at 44.5 degrees C generate a change in the electrical impedance of the culture relative to a sterile control when populations reach 10(6) to 10(7) per ml. The ratio of these changes was measured automatically, and the data were processed by computer. A linear relation was found between the log10 of the number of fecal coliforms in an inoculum and the time required for an electrical impedance ratio signal to be detected. Pure culture inocula consisting of 100 fecal coliforms in log phase or stationary phase were detected in 6.5 and 7.7 h, respectively. Standard curves of log10 fecal coliforms in wastewater inocula versus detection time, based on samples collected at a sewage treatment plant over a 4-month period, were found to vary from one another with time. Nevertheless, detection times were rapid and ranged from 5.8 to 7.9 h for 200 fecal coliforms to 8.7 to 11.4 h for 1 fecal coliform. Variations in detection times for a given number of fecal coliforms were also found among sewage treatment plants. A strategy is proposed which takes these variations into account and allows for rapid, automated enumeration of fecal coliforms in wastewater by the electrical impedance ratio technique.     

Automated electrical impedance technique for rapid enumeration of fecal coliforms in effluents from sewage treatment plants.

Fecal coliforms growing in a selective lactose-based broth medium at 44.5 degrees C generate a change in the electrical impedance of the culture relative to a sterile control when populations reach 10(6) to 10(7) per ml. The ratio of these changes was measured automatically, and the data were processed by computer. A linear relation was found between the log10 of the number of fecal coliforms in an inoculum and the time required for an electrical impedance ratio signal to be detected. Pure culture inocula consisting of 100 fecal coliforms in log phase or stationary phase were detected in 6.5 and 7.7 h, respectively. Standard curves of log10 fecal coliforms in wastewater inocula versus detection time, based on samples collected at a sewage treatment plant over a 4-month period, were found to vary from one another with time. Nevertheless, detection times were rapid and ranged from 5.8 to 7.9 h for 200 fecal coliforms to 8.7 to 11.4 h for 1 fecal coliform. Variations in detection times for a given number of fecal coliforms were also found among sewage treatment plants. A strategy is proposed which takes these variations into account and allows for rapid, automated enumeration of fecal coliforms in wastewater by the electrical impedance ratio technique.     

Enhanced COD and nitrogen removals for the treatment of swine wastewater by combining submerged membrane bioreactor (MBR) and anaerobic upflow bed filter (AUBF) reactor

In order to enhance performances of organics removal and nitrification for the treatment of swine wastewater containing high concentration of organic solids and nitrogen than conventional biological nitrogen removal process, a submerged membrane bioreactor (MBR) was followed by an anaerobic upflow bed filter (AUBF) reactor in this research (AUBF–MBR process). The AUBF reactor is a hybrid reactor, which is the combination of an anoxic filter for denitrification and upflow anaerobic sludge blanket (UASB) for acid fermentation. In the AUBF–MBR process, it showed a considerable enhancement of the effluent quality in terms of COD removal and nitrification. The submerged MBR could maintain more than 14,000 mg VSS/L of the biomass concentration. Total nitrogen (T-N) removal efficiency represented 60% when internal recycle ratio was three times of flow-rate (Q), although the nitrification occurred completely. Although the volatile fatty acids produced in AUBF reactor can enhance denitrification rate, but the AUBF–MBR process showed reduction of overall removal efficiency of the nitrogen due to the reduction of carbon source by methane production in the AUBF reactor compared to that of theoretical nitrogen removal efficiency.

Long-term operation of the submerged MBR showed that the throughputs of the submerged MBR were respectively 74, 63, and 31 days at 10, 15, and 30 L/m² h (LMH) of permeate flux. Resistance to filtration by rejected solid is the primary cause of fouling, however the priority of cake resistance (R_c) and fouling resistance (R_f) with respect to filtration phenomenon was different according to the amount of permeate flux. The submerged MBR, here, achieved a steady-state flux of 15 LMH at 0.4 atm. of trans-membrane pressure (TMP) but the flux can be enhanced in the future because shear force by tangential flow will be greater when multi-layer sheets of membrane were used.     

Removal of micro-organisms in a small-scale hydroponics wastewater treatment system

Aims: To measure the microbial removal capacity of a small-scale hydroponics wastewater treatment plant. Methods and Results: Paired samples were taken from untreated, partly-treated and treated wastewater and analysed for faecal microbial indicators, i.e. coliforms, Escherichia coli, enterococci, Clostridium perfringens spores and somatic coliphages, by culture based methods. Escherichia coli was never detected in effluent water after >5.8-log removal. Enterococci, coliforms, spores and coliphages were removed by 4.5, 4.1, 2.3 and 2.5 log respectively. Most of the removal (60-87%) took place in the latter part of the system because of settling, normal inactivation (retention time 12.7 d) and sand filtration. Time-dependent log-linear removal was shown for spores (k = -0.17 log d(-1), r(2) = 0.99). Conclusions: Hydroponics wastewater treatment removed micro-organisms satisfactorily. Significance and Impact of the Study: Investigations on the microbial removal capacity of hydroponics have only been performed for bacterial indicators. In this study it has been shown that virus and (oo)cyst process indicators were removed and that hydroponics can be an alternative to conventional wastewater treatment.     

Nutrient recovery from domestic wastewater using a UASB-duckweed ponds system

The pilot-scale wastewater treatment system used in this study comprised a 40-l UASB reactor (6-h HRT) followed by three duckweed ponds in series (total HRT 15 days). During the warm season, the treatment system achieved removal values of 93%, 96% and 91% for COD, BOD and TSS, respectively. Residual values of ammonia, TKN and total phosphorus were 0.41 mg N/l, 4.4 mg N/l and 1.11 mg P/l, with removal efficiencies of 98%, 85% and 78%, respectively. The system achieved 99.998% faecal coliform removal during the warm season with final effluent containing 4 x 10(3) cfu/100 ml. During the winter, the system was efficient in removing COD, BOD and TSS but not nutrients. The system was deficient in the removal of faecal coliforms during the winter, producing effluent with 4.7 x 10(5) cfu/100 ml. During the warm season, the N removal consisted of 80% by plant uptake, 5% by sedimentation and 15% unaccounted for. A duckweed production rate of 33 t dry matter per hectare per 8 months was achieved.