Pore‐network modeling of biofilm evolution in porous media

The influence of bacterial biomass on hydraulic properties of porous media (bioclogging) has been explored as a viable means for optimizing subsurface bioremediation and microbial enhanced oil recovery. In this study, we present a pore network simulator for modeling biofilm evolution in porous media including hydrodynamics and nutrient transport based on coupling of advection transport with Fickian diffusion and a reaction term to account for nutrient consumption. Biofilm has non‐zero permeability permitting liquid flow and transport through the biofilm itself. To handle simultaneous mass transfer in both liquid and biofilm in a pore element, a dual‐diffusion mass transfer model is introduced. The influence of nutrient limitation on predicted results is explored. Nutrient concentration in the network is affected by diffusion coefficient for nutrient transfer across biofilm (compared to water/water diffusion coefficient) under advection dominated transport, represented by mass transport Péclet number >1. The model correctly predicts a dependence of rate of biomass accumulation on inlet concentration. Poor network connectivity shows a significantly large reduction of permeability, for a small biomass pore volume. Biotechnol. Bioeng. 2011;108: 2413–2423. © 2011 Wiley Periodicals, Inc.     

Performance evaluation of a newly developed flow diverted bed system for stream restoration

The flow diverted bed (FDB) system was developed to improve the stream water quality. This system (Run 2) had two wetlands where the flow from the upper layer (aerobic) in the first wetland could be redirected to the lower layer (anaerobic) in the second wetland by the flow diverter, and vice versa. In addition, the yellow-soil media placed at the bottom layer allowed a larger influx in the lower layer and had a sorption capacity of phosphate ions. The removal efficiencies of COD and TN at Run 2 [hydraulic loading rate (HLR) = 177 cm/d] was 1.7–3.6 times higher than that at Run 1 (HLR = 192 cm/d) without the diverter. The values of void fraction, dimensionless variance, dispersion number, Peclet number and velocity for this system were found to be 0.6637, 0.2847, 0.1717, 5.8241, and 53.2 m/d, respectively. The reducing trend of each pollutant was exponentially decreased along the overall length of this system. Also, the transport was dominated by convection (plug flow) rather than dispersion (complete mix). The volumetric removal rate constants for COD, SS, TKN, TN, and TP were found to be 4.854, 7.988, 1.388, 1.625, and 1.767 d⁻¹ via the first-order rate model. Consequently, the flow diverter and yellow-soil media in this system played an important role in enhancing the treatability of pollutants and operating at a higher HLR.     

Constructed wetlands for wastewater treatment

The first experiments using wetland macrophytes for wastewater treatment were carried by out by Käthe Seidel in Germany in early 1950s. The horizontal sub-surface flow constructed wetlands (HF CWs) were initiated by Seidel in the early 1960s and improved by Reinhold Kickuth under the name Root Zone Method in late 1960s and early 1970s and spread throughout Europe in 1980s and 1990s. However, cohesive soils proposed by Kickuth got clogged very quickly because of low hydraulic permeability and were replaced by more porous media such as gravel in late 1980s in the United Kingdom and this design feature is still used. In fact, the use of porous media with high hydraulic conductivity was originally proposed by Seidel. HF CWs provide high removal of organics and suspended solids but removal of nutrients is low. Removal of nitrogen is limited by anoxic/anaerobic conditions in filtration beds which do not allow for ammonia nitrification. Phosphorus removal is restricted by the use of filter materials (pea gravel, crushed rock) with low sorption capacity. Various types of constructed wetlands may be combined in order to achieve higher treatment effect, especially for nitrogen. However, hybrid systems are comprised most frequently of vertical flow (VF) and HF systems arranged in a staged manner. HF systems cannot provide nitrification because of their limited oxygen transfer capacity. VF systems, on the other hand, do provide a good conditions for nitrification but no denitrification occurs in these systems. In hybrid systems (also sometimes called combined systems) the advantages of the HF and VF systems can be combined to complement processes in each system to produce an effluent low in BOD, which is fully nitrified and partly denitrified and hence has a much lower total-N outflow concentrations.     

Root architecture and hydraulic conductance in nutrient deprived Pistacia lentiscus L. seedlings

Plants respond to low nutrient availability by modifying root morphology and root system topology. Root responses to nitrogen (N) and phosphorus (P) limitation may affect plant capacity to withstand water stress. But studies on the effect of nutrient availability on plant ability to uptake and transport water are scarce. In this study, we assess the effect of nitrogen and phosphorus limitation on root morphology and root system topology in Pistacia lentiscus L seedlings, a common Mediterranean shrub, and relate these changes to hydraulic conductivity of the whole root system. Nitrogen and phosphorus deprivation had no effect on root biomass, but root systems were more branched in nutrient limited seedlings. Total root length was higher in seedlings subjected to phosphorus deprivation. Root hydraulic conductance decreased in nutrient-deprived seedlings, and was related to the number of root junctions but not to other architectural traits. Our study shows that changes in nutrient availability affect seedling water use by modifying root architecture. Changes in nutrient availability should be taken into account when evaluating seedling response to drought.     

Preliminary media screening for application in the removal of clofibric acid,carbamazepine and ibuprofen by SSF-constructed wetlands

The aim of the present work was to evaluate the sorption capacity of light expanded clay aggregates (LECA) to remove mixtures of ibuprofen, carbamazepine and clofibric acid in water and wastewater. High removal efficiencies were attained for carbamazepine and ibuprofen while a less satisfactory performance was observed for clofibric acid. In a mixture of the three compounds in water a slight decrease in the sorbed amounts is observed in comparison with solutions of the single compounds, indicating some competitive sorption. In wastewater, the pharmaceuticals mixture also undergoes a slight reduction in the sorbed amounts of carbamazepine and ibuprofen, probably due to the presence of dissolved organic matter which increases their solubility. These compounds were removed in the following order of efficiencies in all the tested conditions: carbamazepine > ibuprofen > clofibric acid. Two other clay materials – sepiolite and vermiculite – were tested for the removal of the more recalcitrant clofibric acid, and vermiculite exhibited higher removal efficiency than LECA. The sorption is characterized by an initial fast step, with most pharmaceuticals being removed within the first 24 h. The results of this study are a first step in the process of selecting an appropriate material or combination of materials to be used as media in SSF-CWs designed for the removal of pharmaceuticals from wastewaters.     

Phosphorus retention in lab and field-scale subsurface-flow wetlands treating plant nursery runoff

Constructed wetland systems built to handle nutrient contaminants are often efficient at removing nitrogen, but ineffective at reducing phosphorus (P) loads. Incorporating a clay-based substrate can enhance P removal in subsurface-flow constructed wetland systems. We evaluated the potential of crushed brick, a recycled building product, and two particle sizes of a palygorskite–bentonite industrial mineral aggregate (calcined clay) to sorb P from simulated nutrient-rich plant nursery effluent. The three substrates were screened for P sorbing behavior using sorption, desorption, and equilibration experiments. We selected one substrate to evaluate in an 8-month field trial to compare field sorption capacity with laboratory sorption capacity. In the laboratory, coarse calcined clay average sorption capacity was 497 mg kg⁻¹ and it sorbed the highest percentage of P supplied (76%), except at exposure concentrations >100 mg L⁻¹ where the increased surface area of fine calcined clay augmented its P sorption capacity. Subsurface-flow mesocosms were filled with coarse calcined clay and exposed to a four and seven day hydraulic retention time treatment. Phosphorus export was reduced by 60 to 74% for both treatments until substrate P-binding sites began to saturate during month seven. During the eight month experiment, the four and seven day treatments fixed 1273 ± 22 mg kg⁻¹ P and 937 ± 16 mg kg⁻¹ P, respectively. Sequential extractions of the P saturated clay indicated that P could desorb slowly over time from various pools within the calcined clay; thus, if the calcined clay were recycled as a soil amendment, most P released would be slowly available for plant uptake and use. This study demonstrated the viability of using coarse calcined clay as a root bed substrate in subsurface-flow treatment wetlands remediating phosphorus from plant nursery runoff.     

Culturing Selenastrum capricornutum (Chlorophyta) in a synthetic algal nutrient medium with defined mineral particulates

Algal nutrient studies in chemically-defined media typically employ a synthetic chelator to prevent iron hydroxide precipitation. Micronutrient-particulate interactions may, however, significantly affect chemical speciation and hence biovailability of these nutrients in natural waters. A technique is described by which Selenastrum capricornutum Printz (Chlorophyta) may be cultured in a medium where trace metal speciation (except iron) is controlled, not by organic chelation, but by sorption onto titanium dioxide. Application of this culturing protocol in conjunction with results from sorption studies of nutrient ions on mineral particles provides a means of studying biological impacts of sorptive processes in aquatic environments.     

Effects of aluminium and micro-nutrients on the sorption of phosphorus byTrifolium repens L. cv. ‘Grasslands Huia’ from nutrient solutions during plant induced pH changes

Summary The sorption of phosphorus from nutrient solution and the pH change in the nutrient solution were monitored over a 24 hour period forTrifolium repens L. cv. ‘Grasslands Huia’ plants. Two different concentration levels of micro-nutrients (B, Cu, Fe, Mn and Zn) and Al formed the factors of a fractional replicate of a 2⁶ factorial design. Measurements were made at four time intervals (30 minutes after the plants were placed on the pots, 3 hours, 6 hours and 24 hours later). In addition to phosphorus, fourteen other nutrients (including nitrate and ammonium) were monitored throughout the experiment. The sorption of phosphorus was significantly influenced by both aluminium and iron. The effect of aluminium and iron on phosphorus sorption is attributed to physico-chemical sorption processes involving the root surface. However the effect on the removal of phosphorus by boron, copper, manganese and zinc was observed only as first order interaction effects —B−Zn, Cu−Zn, Mn−Zn. Thus these three elements (B, Cu and Mn) only affect phosphorus removal in conjunction with zinc. Aluminium and iron together had a separate but very significant effect on the removal of phosphorus at most periods throughout the experiment. In contrast, pH was affected only by aluminium, iron (the pH drop was enhanced) and manganese (the pH drop was decreased) as main effects independent of the other treatment elements.     

Leaf litter inputs decrease phosphate sorption in a strongly weathered tropical soil over two time scales

In strongly weathered soils, leaf litter not only returns phosphorus (P) to the soil environment, it may also modify soil properties and soil solution chemistry, with the potential to decrease phosphate sorption and increase plant available P. Using a radioactive phosphate tracer (³²P) and 1 h laboratory incubations we investigated the effect of litter inputs on phosphate sorption over two time scales: (1) long-term field litter manipulations (litter addition, control and litter removal) and (2) pulses of litter leachate (i.e. water extracts of leaf litter) from five species. Leachate pulse effects were compared to a simulated throughfall, which served as a control solution. Soil receiving long-term doubling of leaf litter maintained five-fold more phosphate in solution than the litter removal soil. In addition to the quantity of phosphate sorbed, the field litter addition treatment decreased the strength of phosphate sorption, as evaluated through extraction of sorbed ³²P using a weakly acidic ammonium fluoride solution (Bray 1). In litter removal soil, leachate pulses significantly reduced phosphate sorption in comparison to the throughfall control for all five species evaluated. However, the ability of leachate pulses to reduce phosphate sorption decreased when soil had received field litter inputs. Across soils the effect of leachate pulses on phosphate sorption increased with net sorption of dissolved organic C, with the exception of leachate from one species that had a higher index of aromatic C concentration. These results demonstrate that litter inputs, as both long-term inputs and short-term leachate pulses, can decrease the quantity and strength of phosphate sorption, which may increase the biological availability of this key nutrient.     

Effect of condensate of food waste (CFW) on nutrient removal and behaviours of intercellular materials in a vertical submerged membrane bioreactor (VSMBR)

The main objective of this study was to investigate the effect of condensate of food waste (CFW) on nutrient removal in a pilot-scale vertical submerged membrane bioreactor (VSMBR) treating municipal wastewater having total-chemical oxygen demand to total-nitrogen ratio (T-COD/T-N) of 5.5. In this reactor, the average removal efficiencies of T-COD, T-N, and T-P (total-phosphorus) were 96%, 74%, and 78%, respectively at 8-h hydraulic retention time (HRT), 60-day sludge retention time (SRT), and internal recycle rate of 400%. As the CFW was supplemented with 0.86% of the influent flow rate, the T-N and T-P removal efficiencies increased to 81% and 91%, respectively. Accordingly, in batch tests, it was concluded that the supply of CFW improved enhanced biological phosphorus removal (EBPR) activity of microorganisms resulting in improvement of nutrient removal efficiency. Under this condition, several kinds of poly-hydroxyalkanoates (PHAs) were detected inside the cells.     

Roles of oyster shells in an integrated constructed wetland system designed for P removal

This research aimed to investigate the applicability of an integrated constructed wetland system for P removal from low-strength wastewaters. The integrated system consisted of a constructed wetland and a post-filter unit, in series; both units were packed with oyster shells (OS) as adsorption and filtration media. Based on 1 year of operation under the overall hydraulic retention time of 3.5 days, the integrated system was found to be highly effective in removing BOD₅ (92.3%), N (85.7%), P (98.3%) and total suspended solids (TSS) (94.4%) compounds, in which the constructed wetland unit was responsible for most of the treatment performance, while the post-filter unit served as a polishing unit, especially in the removal of the remaining N, P and TSS. To simulate heavy rainfall conditions, the integrated system was tested under hydraulic shock loading at the overall hydraulic retention time of 0.7 day for 14 days that represented the extreme period of high drainage flows. There were some increases of P concentrations in the post-filter effluent during the 14 days of operation up to about 5 mg/l, but these P concentrations were later decreased to about 1 mg/L after the shock loading period. These experimental results suggested the applicability of the integrated constructed wetland system which used oyster shells as adsorption and filtration media for P removal which should help to minimize eutrophication problems in receiving waters.     

Reliability, repeatability and accuracy of the falling head method for hydraulic conductivity measurements under laboratory conditions

The aim of this study was to verify under lab conditions the reliability, repeatability and accuracy of the falling head method (FHM) for hydraulic conductivity measurements. The FHM is a reliable procedure that has slight variations (less than 10%) in repeated measurements and turns out to be a reliable technique to record the hydraulic conductivities typically described for clogged and unclogged subsurface-flow constructed wetlands (from 4 to ca. 360 m/day). The accuracy of the method is acceptable considering difficulties in the measurement of hydraulic conductivity in highly conductive media. Accordingly, results show measurement deviations of 20% when compared with a laboratory constant head method for highly conductive media (higher than 250 m/day), and 80% for media with low hydraulic conductivity (lower than 50 m/day). The main conclusion of the present paper is that of the FHM is a reliable and repeatable technique for hydraulic conductivity measurements and it is accurate enough for on-site clogging assessment in full-scale constructed wetlands.     

Variation in hydraulic conductivity of mangroves: influence of species, salinity, and nitrogen and phosphorus availability

We investigated how species identity and variation in salinity and nutrient availability influence the hydraulic conductivity of mangroves. Using a fertilization study of two species in Florida, we found that stem hydraulic conductivity expressed on a leaf area basis ( K _leaf) was significantly different among species of differing salinity tolerance, but was not significantly altered by enrichment with limiting nutrients. Reviewing data from two additional sites (Panamá and Belize), we found an overall pattern of declining leaf‐specific hydraulic conductivity ( K _leaf) with increasing salinity. Over three sites, a general pattern emerges, indicating that native stem hydraulic conductivity ( K _h) and K _leaf are less sensitive to nitrogen (N) fertilization when N limits growth, but more sensitive to phosphorus (P) fertilization when P limits growth. Processes leading to growth enhancement with N fertilization are probably associated with changes in allocation to leaf area and photosynthetic processes, whereas water uptake and transport processes could be more limiting when P limits growth. These findings suggest that whereas salinity and species identity place broad bounds on hydraulic conductivity, the effects of nutrient availability modulate hydraulic conductivity and growth in complex ways.     

Primary isolation of Geotrichum klebahnii polygalacturonase by capturing with glass fiber microfilters

This study reports the use of borosilicate glass fiber microfilters (GFMs) for capturing and concentrating Geotrichum klebahnii polygalacturonase (GkPG) from the culture media through a simple coupled filtration–sorption process. GFM was characterized by scanning electron microscopy and intrusion mercury porosimetry. Adsorption of GkPG was almost instantaneous with maximum effectiveness at pH 3.0. Using the parameters of Langmuir model and a material balance, a filter bed consisting of a stack of GFM was designed for recovering GkPG from a certain volume of culture media. After adsorption and washing, the enzyme was efficiently eluted with a buffer solution at pH 5.0 containing 0.5 M NaCl. The filter bed was reused in several sorption cycles and enzyme recovery could be also done without prior cell removal with the benefits of using microfiltration for enzyme cell separation in a single step. Under the process conditions employed, GkPG recovery was about 80% with a concentration factor of about 4-fold.     

Transmural flow bioreactor for vascular tissue engineering

Nutrient transport limitation remains a fundamental issue for in vitro culture of engineered tissues. In this study, perfusion bioreactor configurations were investigated to provide uniform delivery of oxygen to media equivalents (MEs) being developed as the basis for tissue‐engineered arteries. Bioreactor configurations were developed to evaluate oxygen delivery associated with complete transmural flow (through the wall of the ME), complete axial flow (through the lumen), and a combination of these flows. In addition, transport models of the different flow configurations were analyzed to determine the most uniform oxygen profile throughout the tissue, incorporating direct measurements of tissue hydraulic conductivity, cellular O₂ consumption kinetics, and cell density along with ME physical dimensions. Model results indicate that dissolved oxygen (DO) uniformity is improved when a combination of transmural and axial flow is implemented; however, detrimental effects could occur due to lumenal pressure exceeding the burst pressure or damaging interstitial shear stress imparted by excessive transmural flow rates or decreasing hydraulic conductivity due to ME compaction. The model was verified by comparing predicted with measured outlet DO concentrations. Based on these results, the combination of a controlled transmural flow coupled with axial flow presents an attractive means to increase the transport of nutrients to cells within the cultured tissue to improve growth (increased cell and extracellular matrix concentrations) as well as uniformity. Biotechnol. Bioeng. 2009; 104: 1197–1206. © 2009 Wiley Periodicals, Inc.     

Effects of variable hydroperiods and water level fluctuations on denitrification capacity, nitrate removal, and benthic-microbial community structure in constructed wetlands

The hydrologic character of wetlands is one of the attributes by which they are defined. There are, however, conflicting reports about the detrimental versus beneficial responses of wetland systems to water level fluctuations and variable hydroperiods. We conducted water level and hydroperiod fluctuation studies in full-scale experimental wetlands in order to determine the effects of hydraulic operation on wetland performance (in terms of nutrient removal), and benthic-bacterial community function (in terms of denitrification potential, DNP) and structure (via terminal restriction fragment length polymorphisms, T-RFLP). In our comparison, detention time was the controlling factor for nitrate removal at the system level. However, widely fluctuating water levels and variable hydroperiods did not diminish either the nitrate removal capacity of the experimental wetlands, or the size or composition of benthic-bacterial communities relative to the more stable water level systems. Rather, significant differences in denitrification potential rates, bacterial cell densities, and benthic community structure were a function of sampling location within the experimental wetlands regardless of hydraulic operation. The results of this study support the need for reconsidering the hydraulic criteria for wetland delineation.     

Removal of Hexane in Biofilters Packed with Perlite and a Peat–Perlite Mixture

Removal of hexane from air–hexane mixtures in biofilters packed with different solid media under nitrogen supplementation was performed for 70 days. Two columns containing Perlite or a mixture of peat and Perlite, were used. The solid media were supplemented with nitrogen source up to 1 kg/m³ per week for high nutrient supplementation and 0.2 kg/m³ per month for low nutrient supplementation. A high rate of hexane removal: 95 g/m³ h was achieved under high nutrient supplementation, high air flow rate and high hexane concentration. However, the percentage of hexane removal decreased with increasing air flow rate and hexane inlet concentration. For high nutrient supplementation the type of solid medium did not significantly affect the biodegradation capacity. With low nutrient supplementation, the highest removal rate was achieved in the column containing the peat–perlite mixture. The column containing perlite had a significantly lower pressure drop (20 Pa/m) than the 2400–2930 Pa/m observed for the column containing the mixture. Perlite offers an opportunity of running a biofiltration process at a lower and stable pressure drop if the nutrient supplementation is managed properly.     

Converting corn wet-milling effluent into high-value fungal biomass in a biofilm reactor

Rhizopus microsporus was grown in an attached growth system using corn wet-milling effluent as a growth medium. This strain was chosen due to its ability to effectively degrade organic matter in corn wet-milling effluent and for its properties to produce significant levels of protein, chitin and chitosan. Fungal growth and organic removal efficiency were examined under both aseptic and non-aseptic conditions with and without nutrient supplementation. Plastic composite support (PCS) tubes, composed of 50% (w/w) polypropylene (PP) and 50% (w/w) agricultural products were used as support media. Significantly higher organic removal measured as chemical oxygen demand (COD) and biomass yield were observed in the bioreactor with PCS tubes than in two control bioreactors; that is with PP tubes alone and suspended growth (without support media). This confirmed that the PCS support medium with agricultural components enhanced fungal growth and organic removal. The results showed that supplementation of nutrients (e.g., mineral salts) under aseptic conditions enhanced the COD removal from 50% to 55% and observed biomass yield from 0.11 to 0.16 g (dry-weight)/g COD(removed) (i.e., from 0.10 to 0.14 g volatile solids (VS)/g COD(removed) approximately). Non-aseptic operation without nutrient supplementation resulted in an observed biomass yield of 0.32 g volatile suspended solids (VSS)/g COD(removed) with no significant improvement in COD removal ( approximately 53%); whereas with nutrient supplementation, the observed biomass yield increased to 0.56 g VSS/g COD(removed) and COD removal improved to 85%. The fungal system was able to degrade the organic matter with concomitant production of high-value fungal biomass. This is the first study that examined the conversion of corn milling waste stream into high value fungal protein.     

Removal of oil by walnut shell media

Studies were conducted to evaluate the oil sorption capacities of walnut shell media. Sorption capacity is the weight of oil picked up by unit weight of a sorbent. Initial oil pick-up by walnut shell media on pure oil and oil on aqueous medium was evaluated. Batch kinetic studies were conducted to evaluate the equilibrium time required by walnut shell media for sorbing oil. For pure oil medium, sorption capacities of 0.30g/g, 0.51g/g and 0.58g/g were obtained for standard mineral oil, vegetable oil and DoALL Bright-Edge oil, respectively. The results showed sorption capacities of 0.56g/g, 0.58g/g and 0.74g/g for standard mineral oil, vegetable oil and DoALL Bright-Edge oil, respectively, for oil on aqueous medium. It was found that sorbed oil could be recovered from walnut shell media by applying pressure. The study showed that walnut shell media can be used as a sorbent for oil removal.     

Biological and Geochemical Sinks for Phosphorus in Soil from a Wet Tropical Forest

In many tropical and volcanic soils, phosphorus (P) availability is strongly influenced by geochemical sorption, which binds P to soil minerals. The aim of this study was to determine whether biological demand or soil sorption strength was the primary control over phosphate availability and retention in a wet tropical soil with high sorption capacity and low P availability. We added ³²PO₄ to soil from the upper two horizons and assessed the ability of soil microbes to immobilize the added phosphate in the presence of strong sorption. We added phosphate at two concentrations, one representing background turnover that adds low concentrations of P to the soil solution, and the other representing nutrient pulses that can add fairly high fluxes of P to the soil solution. Sorption and microbial immobilization were rapid for both concentrations, consuming most added P within 30 min. Thus, little P remained in the soil solution or extractable pools, which are considered more available to plants. Although soil sorption strength was almost identical for the two horizons, immobilization of tracer P was approximately three times greater in the upper horizon, where most microbial activity was located. This result suggests that microbial demand controlled how P was partitioned into biological versus geochemical sinks. Further evidence for microbial control is suggested by the movement of tracer P from the sorbed pool into the microbial pool when demand was stimulated by the addition of carbon (C). We also explored how increased nitrogen (N) and P availability changed P dynamics in this nutrient poor soil. In contrast to the unfertilized soil, long-term N and P fertilization substantially reduced biological control over inorganic P. P fertilization saturated the soils, overwhelming biological P demand, whereas N fertilization appeared to increase available P through reduced P sorption. Where biological demand for P is high and P becomes available in the soil solution, microbes may play an important role in controlling P partitioning into biological versus geochemical sinks even in soils that have high sorption capacity.