Methanol vapor biofiltration coupled with continuous production of recombinant endochitinase Ech42 by Pichia Pastoris

Methanol biofiltration using methylotrophic microorganisms has been previously reported by various authors. In a previous study, a modified strain of Pichia pastoris was tested for the ability to produce endochitinase (Ech42) when coupled with methanol vapor biodegradation in batch tests. The next challenge was to validate the process in a continuous system. Thus, in the present study, a biofilter packed with perlite and inoculated with P. pastoris transformed with the plasmid pPIC-ech42 was used for methanol vapor biofiltration and the continuous production of recombinant endochitinase (Ech42) for 60 days. The maximum elimination capacity (EC) of methanol obtained was 1320 g m^?3 h^?1 at a loading rate of 1465 g m^?3 h^?1. The extracellular protein production rate in the leachate was 2360 μg h^?1 with a chitinase enzymatic activity of 123 U L^?1. The protein content on the biofilm samples was negligible, indicating the effectiveness of the overall process and of P. pastoris to excrete proteins. The carbon balance indicated that 81% of the consumed methanol was mineralized and 5.8% was incorporated into biomass. The results of this study and the economic balance underscore the promising application of linking methanol vapor biofiltration to the continuous production of recombinant proteins.     

Biotrickling air filtration of 2-chlorophenol at high loading rates

The degradation of 2-chlorophenol vapours in air was performed in a trickling biofilter packed with ceramic material seeded with the bacterium Pseudomonas pickettii, strain LD1. The system performance was evaluated under varying operating conditions (inlet 2-chlorophenol air concentrations from 0.10 to 3.50 g m^?3, and superficial air velocities of 30.0, 60.0, and 120.0 m h^?1). For all air velocity the maximum degradation rate was obtained for loading rates of 40 g m^?2 h^?1. Higher loading conditions resulted in strong inhibition of microbial activity, particularly severe at high air velocity. Process analysis, performed using data on pollutant concentration profiles along the filter packing obtained under different conditions of inlet concentration and air velocity, proves that best performance (i.e. maximum degradation efficiency and capacity) can be obtained for a narrow range of operating conditions, which can be ensured by proper design of biofilter size (i.e. diameter and height). Kinetic analysis of experimental data confirms that 2-CP inhibits microbial activity in the biofilter bed. Experimental data are satisfactorily fitted by the Haldane kinetic equation up to a critical value of loading rate, beyond which the experimental degradation rate is overestimated by the kinetic model. The inhibition appears to be affected by the loading rate, and the estimated inhibition constant linearly increases with increasing empty bed residence time.     

Proposed kinetic mechanism of the production of biodiesel from palm oil using lipase

Experimental determination of the separate effects of palm oil and methanol concentrations on the rate of their enzymatic transesterification was used to propose suitable mechanismic steps and to test the generated kinetic model. The reaction took place in n-hexane organic medium and the lipase used was from Mucor miehei. At a constant methanol concentration of 300 mol m⁻³, it was found that, initially as the palm oil concentration increased, the initial reaction rate increased. However, the initial rate dropped sharply at substrate concentrations larger than 1250 mol m⁻³. Similar behaviour was observed for methanol concentration effect, where at a constant substrate concentration of 1000 mol m⁻³, the initial rate of reaction dropped at methanol concentrations larger than 3000 mol m⁻³. Ping Pong Bi Bi mechanism with inhibition by both reactants was adopted as it best explains the experimental findings. A mathematical model was developed from a proposed kinetic mechanism and was used to identify the regions where the effect of inhibition by both substrates arised. The proposed model equation is essential for predicting the rate of methanolysis of palm oil in a batch or a continuous reactor and for determining the optimal conditions for biodiesel production.     

Nutrient uptake and release in ponds under long-term and short-term lotus (Nelumbo nucifera) cultivation: Influence of compost application

Uptake and release of nutrients from ponds used for lotus cultivation were measured in ponds under short-term (1 yr) cultivation with compost application (pond I) and under long-term (20 yr) cultivation without compost application (pond II). Total inflow loads of TN (irrigation water, rainfall and compost) during lotus cultivation period in ponds I and II were 72.3 and 34.3 kg ha^?1 182 day^?1, respectively. TN removal rates in ponds I and II were 77.3 and 49.8% of total inflow load, respectively. Major removal mechanisms of TN were attributed to microbial processes and uptake by lotus. The total outflow loads (infiltration and runoff) of TN during the lotus cultivation period were 13.9 kg ha^?1 182 day^?1 (19.2% of total inflow TN load) for pond I, and 11.3 kg ha^?1 182 day^?1 (32.9% of total inflow TN load) for pond II. For TP the total inflow loads (irrigation water, rainfall and compost) during lotus cultivation in ponds I and II were 80.8 and 1.9 kg ha^?1 182 day^?1, respectively. TP removal rates in ponds I and II were 84.9 and ?274.1% of total input, respectively. Phosphorus removal was attributed to lotus uptake and soil adsorption. The total outflow loads (infiltration and runoff) of TP during lotus cultivation period were 10.1 kg ha^?1 182 day^?1 (12.5% of total inflow TP load) for pond I, and 6.6 kg ha^?1 182 day^?1 (355.6% of total inflow TP load) for pond II. TN and TP in runoff from pond I (with compost) was higher than that in pond II (without compost), showing that TN and TP in runoff were strongly influenced by compost addition. Therefore, in order to satisfy established water-quality standards, the amount of compost used in lotus cultivation should be evaluated.     

Biological treatment of n-hexane and methanol in trickle bed air biofilters under acidic conditions

This study focuses on evaluating the degradation of n-hexane/methanol mixture in trickle-bed-air-biofilters (TBABs). Two different concentration ratios of methanol:n-hexane were evaluated (3:1) for TBAB “A” and (5:1) for TBAB “B”. Both TBABs were run and fed with nutrients buffered at pH 4 for encouraging the growth of fungi. The TBABs were loaded with pelletized diatomaceous earth support media and were run at an empty bed residence time of 120 s. n-Hexane loading rates (LRs) ranged from 0.9 to 13.2 g/m³ h for both TBABs. The corresponding methanol LRs varied from 2.3 to 37.7 g/m³ h and from 4.6 to 64.5 g/m³ h for TBABs “A” and “B”, respectively. Experimental results have shown that the degradation of n-hexane in presence of methanol is enhanced for n-hexane LRs less than 10.6 g/m³ h as compared to previous study for sole-fed n-hexane, but for n-hexane LRs of 13.2 g/m³ h, the performance of TBABs in eliminating n-hexane depended on the methanol to n-hexane ratios. The impact was less severe for TBAB “A” (RE 85%) as compared to TBAB “B” (RE 72%). This is attributed to the high LRs of methanol in TBAB “B”. n-Hexane performance stability was another advantage attained.     

Nitrification rates of algal–bacterial biofilms in wastewater stabilization ponds under light and dark conditions

The objective of this study was to investigate nitrification rates in algal–bacterial biofilms of waste stabilization ponds (WSP) under different conditions of light, oxygen and pH. Biofilms were grown on wooden plates of 6.0 cm by 8.0 cm by 0.4 cm in a PVC tray continuously fed with synthetic wastewater with initial NH₄-N and Chemical Oxygen Demand (COD) concentrations of 40 mg l^?1 and 100 mg l^?1, respectively, under light intensity of 85–95 μE m^?2 s^?1. Batch activity tests were carried out by exposure of the plates to light conditions as above (to simulate day time), dim light of 1.8–2.2 μE m^?2 s^?1 (to simulate reduced light as in deeper locations in WSP) and dark conditions (to simulate night time). Dissolved oxygen (DO) concentration and pH were controlled. At some experiments, both parameters were kept constant, and at others they were left to vary as in WSP. Results show biofilm nitrification rates of 945–1817 mg-N m^?2 d^?1 and 1124–1615 mg-N m^?2 d^?1 for light and dark experiments. When the minimum DO was 4.1 mg l^?1, the biofilm nitrification rates under light and dark conditions did not differ significantly at 95% confidence. When the minimum DO in the dim light experiment was 3.2 mg l^?1, the nitrification rates under light and dim light conditions were 945 mg-N m^?2 d^?1 and 563 mg-N m^?2 d^?1 and these significantly differed. Further decrease of DO to 1.1 mg l^?1 under dark conditions resulted in more decrease of the nitrification rates to 156 mg-N m^?2 d^?1. It therefore seems that under these experimental conditions, biofilm nitrification rates are significantly reduced at a certain point when bulk water DO is between 3.2 mg l^?1 and 4.1 mg l^?1. As long as bulk water DO under dark is high, light is not important in influencing the process of nitrification.     

Performance of a composite membrane bioreactor treating toluene vapors: Inocula selection,reactor performance and behavior under transient conditions

In this study, a membrane biofilm reactor performance for toluene as a model pollutant is presented. A composite membrane consisting of a porous polyacrylonitrile (PAN) support layer coated with a very thin (0.3 μm) dense polydimethylsiloxane (PDMS) top layer was used. Batch experiments were performed to select an appropriate inocula (slaughterhouse wastewater treatment sludge with a specific toluene consumption rate of 118 ± 23 μg g^?1 VSS L^?1) among the three available sources of inoculums. The maximum elimination capacity gas-side reactor volume based (EC)_v and membrane based (EC)_{m, max} obtained were 609 g m^?3 h^?1 and 1.2 g m^?2 h^?1 respectively, which is much higher than other membrane bioreactors. Further experiments involved the study of the membrane biofilm reactor flexibility when operational parameters as temperature, loading rate etc. were modified. In all cases, the membrane biofilm reactor showed a rapid adaptation and new steady-states were obtained within hours. Overall, the results illustrate that membrane bioreactors can potentially be a good option for treatment of air pollutants such as toluene.     

The seed bank in a subtropical freshwater marsh: implications for wetland restoration

Seed bank samples were collected from Huli Marsh, a subtropical shallow water mountainous marsh in Hunan Province, South China. Core samples were divided into upper and lower layers (each 5 cm in depth) and allowed to germinate in three water levels (0, 5 and 10 cm) over a 4-month period. A total of 51 species germinated and the mean density was 9211 ± 7188 seedlings m⁻². In the top 5 cm 41 species and 5747 ± 5111 seedlings m⁻² germinated, whereas 40 species and 3464 ± 3363 seedlings m⁻² did so from 5–10 cm. Germinated seedling density was significantly higher in the upper layer, largely due to differences in eight species. With increasing experimental water depth, less seedlings germinated: respectively, 9788 ± 7157 m⁻², 2050 ± 2412 m⁻² and 1978 ± 2616 m⁻², of 44, 21 and 19 species, submerged under 0, 5 or 10 cm. Seven species could emerge only in 0 water level. Vallisneria natans occurred only in 5 cm water, whereas Ottelia alismoides occurred in 10 cm water. In the vegetation survey of the marsh, 25 species were recorded, which was less than half of the species recorded in the seed bank. The top 10 dominants in the standing vegetation, accounting for 89% of vegetation abundance, represented only 10% in the seed bank. Twenty germinated species that also occurred in the standing vegetation accounted for 56% of the total seed bank. Our observed number of species germinating from a Chinese wetland seed bank is within the range observed elsewhere in the northern hemisphere (15–113 species).     

Colonization dynamics of biofilm-associated ciliate morphotypes at different flow velocities

The impact of flow velocity on initial ciliate colonization dynamics on surfaces were studied in the third order Ilm stream (Thuringia, Germany) at a slow flowing site (0.09 m s^?1) and two faster flowing sites (0.31 m s^?1) and in flow channels at 0.05, 0.4, and 0.8 m s^?1. At the slow flowing stream site, surfaces were rapidly colonized by ciliates with up to 60 cells cm^?2 after 24 h. In flow channels, the majority of suspended ciliates and inorganic matter accumulated at the surface within 4.5 h at 0.05 m s^?1. At 0.4 m s^?1 the increase in ciliate abundance in the biofilm was highest between 72 and 168 h at about 3 cells cm^?2 h^?1. Faster flow velocities were tolerated by vagile flattened ciliates that live in close contact to the surface. Vagile flattened and round filter feeders preferred biofilms at slow flow velocities. Addition of inorganic particles (0, 0.6, and 7.3 mg cm^?2) did not affect ciliate abundance in flow channel biofilms, but small ciliate species dominated and number of species was lowest (16 species cm^?2) in biofilms at high sediment content. Although different morphotypes dominated the communities at contrasting flow velocities, all functional groups contributed to initial biofilm communities implementing all trophic links within the microbial loop.     

Nitrate-nitrogen retention in wetlands in the Mississippi River Basin

Nitrate-nitrogen retention as a result of river water diversions is compared in experimental wetland basins in Ohio for 18 wetland-years (9 years × 2 wetland basins) and a large wetland complex in Louisiana (1 wetland basin × 4 years). The Ohio wetlands had an average nitrate-nitrogen retention of 39 g-N m⁻² year⁻¹, while the Louisiana wetland had a slightly higher retention of 46 g-N m⁻² year⁻¹ for a similar loading rate area. When annual nitrate retention data from these sites are combined with 26 additional wetland-years of data from other wetland sites in the Basin Mississippi River (Ohio, Illinois, and Louisiana), a robust regression model of nitrate retention versus nitrate loading is developed. The model provides an estimate of 22,000 km² of wetland creation and restoration needed in the Mississippi River Basin to remove 40% of the nitrogen estimated to discharge into the Gulf of Mexico from the river basin. This estimated wetland restoration is 65 times the published net gain of wetlands in the entire USA over the past 10 years as enforced by the Clean Water Act and is four times the cumulative total of the USDA Wetland Reserve Program wetland protection and restoration activity for the entire USA.     

Influence of urea and calcium dosage on the effectiveness of bacterially induced carbonate precipitation on limestone

Bacterially induced carbonate precipitation has been explored for the protection and consolidation of ornamental stone. Attempts to improve the efficiency of this biodeposition process were primarily focused on the microbial aspects, i.e. type of microorganism and metabolic pathway. In this study, the influence of the chemical parameters, i.e. concentration of calcium salts and urea, on the effectiveness of the biodeposition treatment has been examined. The amount of calcium carbonate that can be precipitated in the stone is conditioned both by the amount of cells retained in the stone and the concentration of urea and calcium used. From sonication experiments, a good consolidation was observed for limestone prisms treated with a calcium dosage of 17 g Ca²⁺ m^?2 with no improvement at higher concentrations. For limestone prisms of 4 cm × 2 cm × 1 cm, the biodeposition treatment resulted in a 63% lower weight loss upon sonication compared to untreated specimens. The waterproofing effect was observed to increase with increasing calcium dosages. While for a calcium dosage of 17 g Ca²⁺ m^?2 the water absorption was similar to that of untreated specimens, concentrations of 67 g Ca²⁺ m^?2 resulted in a 50% decrease of the rate of water absorption. For calcium dosages higher than 34 g Ca²⁺ m^?2 a significant change in the visual aspect (ΔE > 6) of the treated stones could be observed. Overall, the urea/calcium chloride-based biodeposition treatment attained a protective performance comparable with that of the commonly used ethylsilicates.     

The effect of internal capacitance on power quality and energy efficiency in a tubular microbial fuel cell

The pseudo-capacitive behaviour of a high surface area carbon veil electrode in a tubular microbial fuel cell (MFC) was investigated as a mechanism to enhance power quality and energy efficiency. Accumulated charge and energy from the anodic biofilm after prolonged open circuit times (1–120 min) were compared against equivalent periods of steady state loading (R = 100–3000 Ω). A significant difference in the amount of accumulated charge with different loads was observed, resulting in 1.051 C (R = 100 Ω) compared to 0.006 C (R = 3 kΩ). The automated application of short open and closed circuit (0.5–10 s) cycles resulted in an increase of power/current production (closed circuit alone), but presented lower efficiency considering entire open and closed period. The cumulative charge on the carbon veil electrode with biofilm was 39,807 C m⁻² at 100 Ω. Electrochemical Impedance Spectroscopy (EIS) showed that the Helmholtz layer presented a double layer capacitance of more than ten times the biofilm on electrode. The results indicate that the capacitive behaviour could be utilized to increase the power quality, i.e. its availability/applicability with respect to the operation of low power consuming devices.     

Continuous succinic acid fermentation by Actinobacillus succinogenes

Fermentations were performed in an external recycle bioreactor using CO₂ and d-glucose at feed concentrations of 20 and 40 g L⁻¹. Severe biofilm formation prevented kinetic analysis of suspended cell (‘chemostat’) fermentation, while perlite packing enhanced the volumetric productivity by increasing the amount of immobilised cells. The highest productivity of 6.35 g L⁻¹ h⁻¹ was achieved at a dilution rate of 0.56 h⁻¹. A constant succinic acid yield of 0.69 ± 0.02 g/(g of glucose consumed) was obtained and found to be independent of the dilution rate, transient state and extent of biofilm build-up – approximately 56% of the carbon that formed phosphoenolpyruvate ended up as succinate. Byproduct analysis indicated that pyruvate oxidation proceeded solely via the formate-lyase pathway. Cell growth and corresponding biofilm formation were rapid at dilution rates higher than 0.35 h⁻¹ when the product concentrations were low (succinic acid < 10 g L⁻¹), while minimal growth was observed at succinic acid concentrations above this threshold.     

Performance evaluation and effects of hydraulic retention time and mass loading rate on treatment of woodwaste leachate in surface-flow constructed wetlands

Four surface-flow mesocosm wetlands were operated at different hydraulic retention times during two periods to treat diluted woodwaste leachate that was acidic, of very high oxygen demand, and toxic. Temperature, dissolved oxygen, and redox potential decreased with increasing water depth. However, there was no significant vertical variation in microbial biomass. No significant development in biomass of planktonic microorganisms was found over 6 weeks of initial operation. It took <1–6 weeks for maturation of the biofilm on submerged plant surfaces and the sedimentary microbial community. Mass reduction efficiencies of chemical oxygen demand, and tannin and lignin increased significantly with hydraulic retention time when 10% leachate was fed with tap water. When a more recalcitrant influent was fed, there was a slight increase of reduction efficiency with increasing hydraulic retention time. Reduction rates increased linearly with mass loading rates up to 0.4 kg m⁻³ d⁻¹ chemical oxygen demand and 0.13 kg m⁻³ d⁻¹ tannin and lignin. Precipitation and evapotranspiration had profound impacts on the overall performance and its variability. Mass balance-based operating data of wetlands with a mature microbial community are required for proper performance assessment.     

Treatment of wet process hardboard plant VOC emissions by a pilot scale biological system

A pilot scale biological treatment system for air emissions was installed and tested at a forest products plant in western Oregon, USA, which collected and treated gaseous emissions from the hardboard steam press vents on the top of the plant building. This system was installed mainly to demonstrate the effectiveness of biological treatment technologies in removing volatile organic compounds (VOC) and hazardous air pollutants (HAP) from the wet-process hardboard press emissions, and to test the efficiency of the system on fine particles and condensable organics with the presence of a pre-treatment wet dust collector. The bio-oxidation system was comprised of a particle pre-treatment unit Type W Rotoclone (wet hydrocyclone), a biotrickling filter and a biofilter with airflow capacity of up to 4.72 m³/s. This unit operated at approximately 0.71 m³/s, which is the optimal flow required for the Rotoclone's throughput, and provided an EBCT (empty bed contact time) of 45 s. Analysis of total VOC measurements from the system indicated removals down to less than 5 ppm in the effluent emissions. Evaluations of opacity reductions also met project objectives with routine outlet measurements of 0–5%, which are in compliance with state regulatory guidelines. Emissions air samples were collected at different locations in the biological system for GC–MS analysis and characterization to identify specific VOCs and their removals.     

Diurnal exchanges of CO2 and CH4 across the water–atmosphere interface in a water chestnut meadow (Trapa natans L.)

CH₄ and CO₂ fluxes across the water–atmosphere interface were measured over a 24 h day–night cycle in a shallow oxbow lake colonized by the water chestnut (Trapa natans L.) (Lanca di Po, Northern Italy). Only exchanges mediated by macrophytes were measured, whilst gas ebullition was not considered in this study. Measurements were performed from 29 to 30 July 2005 with short incubations, when T. natans stands covered the whole basin surface with a mean dry biomass of 504 ± 91 g m⁻². Overall, the oxbow lake resulted net heterotrophic with plant and microbial respiration largely exceeding carbon fixation by photosynthesis. The water chestnut stand was a net sink of CO₂ during the day-light period (−60.5 ± 8.5 mmol m⁻² d⁻¹) but it was a net source at night (207.6 ± 6.1 mmol m⁻² d⁻¹), when the greatest CO₂ efflux rate was measured across the water surface (28.2 ± 2.4 mmol m⁻² h⁻¹). The highest CH₄ effluxes (6.6 ± 1.8 mmol m⁻² h⁻¹) were determined in the T. natans stand during day-time, whilst CH₄ emissions across the plant-free water surface were greatest at night (6.8 ± 2.1 mmol m⁻² h⁻¹). Therefore, we assumed that the water chestnut enhanced methane delivery to the atmosphere. On a daily basis, the oxbow lake was a net source to the atmosphere of both CO₂ (147.1 ± 10.8 mmol m⁻² d⁻¹) and CH₄ (116.3 ± 8.0 mmol m⁻² d⁻¹).     

Estimation of big sagebrush leaf area index with terrestrial laser scanning

Accurate monitoring and quantification of the structure and function of semiarid ecosystems is necessary to improve carbon and water flux models that help describe how these systems will respond in the future. The leaf area index (LAI, m² m⁻²) is an important indicator of energy, water, and carbon exchange between vegetation and the atmosphere. Remote sensing techniques are frequently used to estimate LAI, and can provide users with scalable measurements of vegetation structure and function. We tested terrestrial laser scanning (TLS) techniques to estimate LAI using structural variables such as height, canopy cover, and volume for 42 Wyoming big sagebrush (Artemisia tridentata subsp. wyomingensis Beetle & Young) shrubs across three study sites in the Snake River Plain, Idaho, USA. The TLS-derived variables were regressed against sagebrush LAI estimates calculated using specific leaf area measurements, and compared with point-intercept sampling, a field method of estimating LAI. Canopy cover estimated with the TLS data proved to be a good predictor of LAI (r² = 0.73). Similarly, a convex hull approach to estimate volume of the shrubs from the TLS data also strongly predicted LAI (r² = 0.76), and compared favorably to point-intercept sampling (r² = 0.78), a field-based method used in rangelands. These results, coupled with the relative ease-of-use of TLS, suggest that TLS is a promising tool for measuring LAI at the shrub-level. Further work should examine the structural measures in other similar shrublands that are relevant for upscaling LAI to the plot-level (i.e., hectare) using data from TLS and/or airborne laser scanning and to regional levels using satellite-based remote sensing.     

Effects of irradiance and water flow on formation and growth of spongy and filamentous thalli of Codium fragile

Effects of irradiance and water flow on formation and growth of filamentous and spongy thalli of Codium fragile (Suringar) Hariot growing on vinylon threads were investigated at the laboratory culture. They showed clear differences in their irradiance and water flow requirements for their formation and growth. Spongy thalli were formed from the cultured filamentous thalli only at the high water flow velocity (10 cm s⁻¹). Number of the spongy thalli remarkably increased with increasing irradiance because those at 10, 50 and 100 μmol m⁻² s⁻¹ reached 0, 2 and 76 thalli m⁻¹, respectively, by 10 weeks of culture. In contrast, filamentous thalli were formed from the cultured spongy thalli at 0 and 3 cm s⁻¹, and difference in irradiance had no effect on their formation. Growth of the spongy thalli greatly accelerated under the combination of the high irradiance and high water velocity (200 μmol m⁻² s⁻¹ and 10 cm s⁻¹) because their relative growth rate in wet weight under the condition was two–four times higher than those at the other examined irradiances and water velocities. On the other hand, difference in water velocity had no effect on growth of the filamentous thalli under flowing water, and their growth decelerated at the high irradiance (200 μmol m⁻² s⁻¹). This demonstrates that water flow is a major factor controlling the formation of the spongy and filamentous thalli. The formation and growth of the spongy thalli surely occur under the combination of the high irradiance and fast flowing water. In contrast, the formation of the filamentous thalli occurs in the calm water, and their growth is inhibited under the high irradiance.     

Effects of wetland construction on water quality in a semi-arid catchment degraded by intensive agricultural use

Many factors can influence the improvement of water quality in surface-flow constructed wetlands (SFW). To test if water quality was improved, especially in nutrient and salt content, after passage through SFW, 11 wetland plots of various sizes (50, 200, 800 and 5000 m²) were established within constructed wetlands on agricultural soils in the Ebro River basin (NE Spain) that had been affected by salinization. A set of 15 water quality parameters (e.g., nutrients, salts, sediments, and alkalinity) was obtained from samples collected at the inflow and outflow of the wetlands during the first 4 years after the wetlands were constructed. NO₃-N retention rates were as high as 99% in the largest (5000 m²) wetlands. After 4 years, total phosphorus was still being released from the wetlands but not salts. Over the same period, in small wetlands (50, 200, and 800 m²), retention rate relative to the input of NO₃-N increased from 40% to almost 60%. Retention of NO₃-N amounted to up to 500 g N m^?2 per year, for an average load concentration at inflow of ～20 mg l^?1. Release of Na⁺ declined from 16% to 0–2% by volume, for an average load concentration at inflow of ～70 mg l^?1. At the current retention rate of NO₃-N (76–227 g m^?2 per year), 1.5–4% of the catchment should be converted into wetlands to optimize the elimination of NO₃-N.     

Treatment of high-strength wastewater in tropical vertical flow constructed wetlands planted with Typha angustifolia and Cyperus involucratus

The ability of vertical flow (VF) constructed wetland systems to treat high-strength (ca. 300 mg L^?1 of COD and ca. 300 mg L^?1 total-nitrogen) wastewater under tropical climatic conditions was studied during a 5-month period. Nine 0.8-m diameter experimental VF units (depth 0.6 m) were used: three units were planted with Typha angustifolia L., another three units were planted with Cyperus involucratus Rottb and three units were unplanted. Each set of units were operated at hydraulic loading rates (HLRs) of 20, 50 and 80 mm d^?1. Cyperus produced more shoots and biomass than the Typha, which was probably stressed because of lack of water. The high evapotranspirative water loss from the Cyperus systems resulted in higher effluent concentrations of COD and total-P, but the mass removal of COD did not differ significantly between planted and unplanted systems. Average mass removal rates of COD, TKN and total-P at a HLR of 80 mm d^?1 were 17.8, 15.4 and 0.69 g m^?2 d^?1. The first-order removal rate constants at a HLR of 80 mm d^?1 for COD, TKN and total-P were 49.8, 30.1 and 13.5 m year^?1, respectively, which is in the higher range of k-values reported in the literature. The oxygen transfer rates were ca. 80 g m^?2 d^?1 in the planted systems as opposed to ca. 60 g m^?2 d^?1 in the unplanted systems. The number of Nitrosomonas was two to three orders of magnitude higher in the planted systems compared to the unplanted systems. Planted systems thus had significantly higher removal rates of nitrogen and phosphorus, higher oxygen transfer rates, and higher quantities of ammonia-oxidizing bacteria. None of the systems did, however, fully nitrify the wastewater, even at low loading rates. The vertical filters did not provide sufficient contact time between the wastewater and the biofilm on the gravel medium of the filters probably because of the shallow bed depth (0.6 m) and the coarse texture of the gravel. It is concluded that vertical flow constructed wetland systems have a high capacity to treat high-strength wastewater in tropical climates. The gravel and sand matrix of the vertical filter must, however, be designed in a way so that the pulse-loaded wastewater can pass through the filter medium at a speed that will allow the water to drain before the next dose arrives whilst at the same time holding the water back long enough to allow sufficient contact with the biofilm on the filter medium.