Rapid and direct estimation of active biomass on granular activated carbon through adenosine tri-phosphate (ATP) determination

Granular activated carbon (GAC) filtration is used during drinking water treatment for the removal of micropollutants such as taste and odour compounds, halogenated hydrocarbons, pesticides and pharmaceuticals. In addition, the active microbial biomass established on GAC is responsible for the removal of biodegradable dissolved organic carbon compounds present in water or formed during oxidation (e.g., ozonation and chlorination) processes. In order to conduct correct kinetic evaluations of DOC removal during drinking water treatment, and to assess the state and performance of full-scale GAC filter installations, an accurate and sensitive method for active biomass determination on GAC is required. We have developed a straight-forward method based on direct measurement of the total adenosine tri-phosphate (ATP) content of a GAC sample and other support media. In this method, we have combined flow-cytometric absolute cell counting and ATP analysis to derive case-specific ATP/cell conversion values. In this study, we present the detailed standardisation of the ATP method. An uncertainty assessment has shown that heterogeneous colonisation of the GAC particles makes the largest contribution to the combined standard uncertainty of the method. The method was applied for the investigation of biofilm formation during the start-up period of a GAC pilot-scale plant treating Lake Zurich water. A rapid increase in the biomass of up to 1.1 x 10(10)cells/g GAC dry weight (DW) within the first 33 days was observed, followed by a slight decrease to an average steady-state concentration of 7.9 x 10(9)cells/g GAC DW. It was shown that the method can be used to determine the biomass attached to the GAC for both stable and developing biofilms.     

Development of biomass in a drinking water granular active carbon (GAC) filter

Indigenous bacteria are essential for the performance of drinking water biofilters, yet this biological component remains poorly characterized. In the present study we followed biofilm formation and development in a granular activated carbon (GAC) filter on pilot-scale during the first six months of operation. GAC particles were sampled from four different depths (10, 45, 80 and 115 cm) and attached biomass was measured with adenosine tri-phosphate (ATP) analysis. The attached biomass accumulated rapidly on the GAC particles throughout all levels in the filter during the first 90 days of operation and maintained a steady state afterward. Vertical gradients of biomass density and growth rates were observed during start-up and also in steady state. During steady state, biomass concentrations ranged between 0.8-1.83 x 10⁻⁶ g ATP/g GAC in the filter, and 22% of the influent dissolved organic carbon (DOC) was removed. Concomitant biomass production was about 1.8 × 10¹² cells/m²h, which represents a yield of 1.26 × 10⁶ cells/μg. The bacteria assimilated only about 3% of the removed carbon as biomass. At one point during the operational period, a natural 5-fold increase in the influent phytoplankton concentration occurred. As a result, influent assimilable organic carbon concentrations increased and suspended bacteria in the filter effluent increased 3-fold as the direct consequence of increased growth in the biofilter. This study shows that the combination of different analytical methods allows detailed quantification of the microbiological activity in drinking water biofilters.     

Nutrient gradients in a granular activated carbon biofilter drives bacterial community organization and dynamics

The quality of drinking water is ensured by hygienic barriers and filtration steps, such as ozonation and granular activated carbon (GAC) filtration. Apart from adsorption, GAC filtration involves microbial processes that remove biodegradable organic carbon from the ozonated ground or surface water and ensures biological stability of the treated water. In this study, microbial community dynamics in were monitored during the start-up and maturation of an undisturbed pilot-scale GAC filter at 4 depths (10, 45, 80 and 115 cm) over a period of 6 months. New ecological tools, based on 16S rRNA gene-DGGE, were correlated to filter performance and microbial activity and showed that the microbial gradients developing in the filter was of importance. At 10 cm from the top, receiving the freshly ozonated water with the highest concentration of nutrients, the microbial community dynamics were minimal and the species richness remained low. However, the GAC samples at 80-115 cm showed a 2-3 times higher species richness than the 10-45 cm samples. The highest biomass densities were observed at 45-80 cm, which corresponded with maximum removal of dissolved and assimilable organic carbon. Furthermore, the start-up period was clearly distinguishable using the Lorenz analysis, as after 80 days, the microbial community shifted to an apparent steady-state condition with increased evenness. This study showed that GAC biofilter performance is not necessarily correlated to biomass concentration, but rather that an elevated functionality can be the result of increased microbial community richness, evenness and dynamics.     

Accumulation and fate of green fluorescent labeled Escherichia coli in laboratory-scale drinking water biofilters

Biological filters combining microbial activity and rapid sand filtration are used in drinking water treatment plants for enhanced biodegradable organic matters (BOM) removal. Biofilms formed on filter media comprised of bacteria enclosed in a polymeric matrix are responsible for the adsorption of BOM and attachment of planktonic microorganisms. This study investigated the removal of Escherichia coli cells injected into laboratory-scale biofilters and the role of biofilm in retaining the injected E. coli. Green fluorescent protein was used as a specific marker to detect and quantify E. coli in the biofilms. About 35% of the total injected E. coli cells were observed in the filter effluents, when initial cell concentrations were measured at 7.4 x 10(6) CFU/mL and 1.6 x 10(7) CFU/mL in two separate experiments. The results from real-time PCR and plate count analysis indicated that 95% of the E. coli retained inside the filters were either non-viable or could not be recovered by colony counting techniques. Injected cells were unevenly distributed inside the filter with more than 70% located at the top 1/5 of the filter. Images obtained from an epifluorescent microscope showed that E. coli cells were embedded inside the biofilm matrix and presented mainly as microcolonies intertwined with other microorganisms, which was consistent with findings from standard plate count methods and qPCR.     

饮用水生物强化过滤工艺生物膜特性研究

以水厂沉淀池出水为原水,对生物强化过滤工艺的生物膜形成过程进行了研究,探讨了不同滤料介质组成的生物滤柱的生物膜特性,分析了膜形成过程中污染物的去除效果和滤柱生物量的变化情况,并对膜形成过程的影响因素进行了讨论.结果表明,在活性炭-石英砂滤料上生物膜形成效果要优于无烟煤-石英砂双层滤料和石英砂单层滤料;反冲洗水含氯对生物膜形成有负面影响,对无烟煤-石英砂滤柱的影响尤为显著;可以CODMn和NO-2的去除率作为生物膜成熟的评价指标.     

Indicators of biofilm development and activity in constructed wetlands microcosms

Methods to measure protein, exopolysaccharide, viable cell number and INT reduction activity were tested on biofilm growing in a wastewater batch reactor. They were shown to be meaningful indicators of biofilm growth and correlated well with each other. Protein, exopolysaccharide, viable cells and INT reduction rates increased linearly over time. Viable cell number exhibited strong linear correlations with protein (R2= 0.98) and exopolysaccharide (R2= 0.99) while INT reduction rate was somewhat less well correlated (R2= 0.90). Our results indicate production rates of 0.91 x 10(-7) microg EPS per viable cell and 1.0 x 10(-7) microg protein per viable cell. Protein and polysaccharide specific INT reduction rates decreased by approximately 50%, whereas viable cell specific INT reduction rates decreased by 65% and the protein to polysaccharide ratio stayed relatively constant at between 1.1 and 1.2 as the biofilm developed. Measurement of protein, polysaccharide, viable cells and INT reduction rate at depth within the bioreactor showed that they were concentrated in the top 1cm of the influent end of the reactor and each decreased to a base level within 4.5 cm of the inlet. Protein to polysaccharide ratios increased with depth in the reactor and the specific INT reduction rates were maximal at 4.5 cm depth. The results indicate that the biomass can take upwards of 100 days to stabilize during batch (fill and draw) operation of subsurface wetlands and that the relative ratios of biomass components remain relatively constant during biofilm growth. Also, it appears that filtration of suspended solids results in biomass concentration at the inlet to the wetland.     

Biofilm formation and multiplication of Legionella in a model warm water system with pipes of copper, stainless steel and cross-linked polyethylene

Legionella pneumophila was grown in a model warm water system with pipes of copper (Cu), stainless steel (SS) and cross-linked polyethylene (PEX) during recirculation of tap water at 25--35 degrees C. Subsequently, domestic use of warm (37 degrees C) water was simulated using tap water with a low AOC concentration (<10 microg C/L). Two times each week the temperature of the water in the electric heaters (not in the pipes) was elevated to 70 degrees C for 30 min. ATP concentrations in the water sampled from the pipes over a 2-year period were significantly different for the pipe materials, with median values of 2.1 ng/l (Cu), 2.5 ng/l (SS) and 4.5 ng/l (PEX), respectively. Median values of the biofilm concentration were similar on Cu and SS (about 630 pg ATP/cm(2)) and 1870 pg ATP/cm(2) on PEX. Legionella multiplied in these biofilms and median values of Legionella concentrations in water were 1500 CFU/l (Cu) and about 4300 CFU/l for SS and PEX. Legionella to ATP ratios in water had median values of about 0.8 CFU/pg. Hot water flushing (70 degrees C) of the pipes on day 552, followed by 2 weeks of recirculation at 37 degrees C, caused strongly increased concentrations of ATP (up to 300 ng/l) and Legionella (>10(7)CFU/l), with about 100 CFU/pg ATP. Concentrations declined to original levels within 1 week of domestic water use, etc. Legionella concentrations in water and biofilms were at the same levels for all materials after 2 years. Hence, copper temporarily limited the growth of Legionella under the applied conditions and a rapid biomass development strongly increased the Legionella to ATP ratio.     

A comparison of media types in acetate fed expanded-bed anaerobic reactors

A synthetic feed, containing acetate as the only carbon source, was used to start-up four different anaerobic expanded-bed reactors containing three different types of microbial attachment media. The media types used were low-density anthracite, granular activated carbon (GAC) and two sizes of sand. All media types were of the same average diameter, 0.7 mm, except for a smaller sand, 0.35 mm. These media types were chosen to compare surface roughness, macroscopic shear stresses due to upflow velocity and sphericity. The 0.7 mm sand required the greatest upflow velocity, 16 cm/s, while the other reactors had upflow velocities of 5.5–6.0 cm/s. Sand had the least surface roughness and GAC had the roughest surface, while anthracite had the most angular shape. At steady-state, the GAC reactor retained 3.75–10 times the attached biomass retained on the other media tested and the GAC reactor accumulated biomass at a faster rate during start-up. Shear losses reflected the biomass accumulation with the two sand and anthracite media having shear loss coefficients 6–20 times greater than that of the GAC medium. Sand induced the formation of sludge granules in both sand reactors with two species of methanogens and stability of the sludge blankets was critical to reactor performance. Scanning electron microscopy demonstrated that attached growth developed in crevices where biomass was protected from shear forces. Attached growth on the sand and anthracite media was located only in crevices, while the GAC medium is completely covered with crevices and biofilm developed on the entire GAC particle. Surface roughness was critical to biofilm development with the rougher surface providing the better attachment medium.     

Biomass development in slow sand filters

Microbial biomass development in the sand and schmutzdecke layer was determined in two full-scale slow sand filters, operated with and without a light excluding cover. A standard chloroform fumigation-extraction technique was adapted to routinely measure microbial biomass concentrations in the sand beds. Sand was sampled to a depth of 10 cm and schmutzdecke was also collected at the same random positions on the uncovered filter. Interstitial microbial biomass in the uncovered sand bed increased with time and decreased with sampling depth. There was a small accumulation of sand biomass with time in the covered filter, but no relationship was apparent between biomass concentration and depth in this filter. Schmutzdecke did not develop on the covered filter and was spatially highly variable in the uncovered condition compared to the consistent patterns observed in interstitial biomass production. It is speculated that microbial biomass in the sand of uncovered filters is largely related to carbon inputs from photosynthetic activity in the schmutzdecke and involves mechanisms that spatially distribute carbon substrate from the schmutzdecke to the sand. However, total organic carbon and dissolved organic carbon removals were similar in both filters suggesting that relatively small biomass populations in covered filters are sufficient to remove residual labile carbon during advanced water treatment and little further advantage to water purification and organic carbon removal is gained by the increased production of biomass in uncovered slow sand filter beds.     

Role of predation by zooplankton in transport and fate of protozoan (oo)cysts in granular activated carbon filtration

The significance of zooplankton in the transport and fate of pathogenic organisms in drinking water is poorly understood, although many hints of the role of predation in the persistence of microorganisms through water treatment processes can be found in literature. The objective of this study was to assess the impact of predation by natural zooplankton on the transport and fate of protozoan (oo)cysts in granular activated carbon (GAC) filtration process. UV-irradiated unlabelled Cryptosporidium parvum and Giardia lamblia (oo)cysts were seeded into two pilot-scale GAC filtration columns operated under full-scale conditions. In a two-week period after seeding, a reduction of free (oo)cysts retained in the filter bed was observed. Zooplankton was isolated from the filter bed and effluent water on a 30 μm net before and during the two-week period after seeding; it was enumerated and identified. Rotifers, which are potential predators of (oo)cysts, accounted for the major part of the isolated zooplankton. Analytical methods were developed to detect (oo)cysts internalized in natural zooplankton isolated from the filter bed and effluent water. Sample sonication was optimized to disrupt zooplankton organisms and release internalized microorganisms. (Oo)cysts released from zooplankton after sonication were isolated by IMS and stained (EasyStain™) for microscopic counting. Both Cryptosporidium and Giardia (oo)cysts were detected in association with zooplankton in the filter bed samples as well as in the effluent of GAC filters. The results of this study suggest that predation by zooplankton can play a role in the remobilization of persistent pathogens such as Cryptosporidium and Giardia (oo)cysts retained in GAC filter beds, and consequently in the transmission of these pathogens in drinking water.     

Effect of NOM and biofilm on the removal of Cryptosporidium parvum oocysts in rapid filters

Laboratory experiments were performed to evaluate the effects of biofilm and natural organic matter (NOM) on removal of Cryptosporidium parvum oocysts from water by filtration. The bench-scale rapid filters consisted of 2.54 cm ID x 30.5 cm polycarbonate plastic columns packed with 0.55 mm spherical glass beads to a depth of 25 cm and a porosity of 40%. Calcium chloride (0.01 M) served as the coagulant in most of the experiments. The oocyst removal efficiency decreased from 51 +/- 6% for a clean bed to 23 +/- 3% for the biofilm-coated bed and to 14 +/- 1% in the presence of 5 ppm of NOM. The oocyst removal for an experiment with a combination of biofilm-coated filter media and NOM was similar to that for the experiment with NOM alone (15 +/- 1%). The zeta potential values for the oocysts pre-equilibrated with NOM were significantly more negative than those obtained for untreated oocysts. This suggests that NOM enhanced the electrostatic repulsion between the oocysts and the negatively charged glass beads. Fortunately, use of alum as coagulant at a dosage sufficient to neutralize the surface charge of the NOM-coated oocysts resulted in a high removal efficiency (73 +/- 6%). Pre-equilibration of the oocysts with NOM also increased the hydrophobicity of the oocysts, but this was deemed to have a negligible effect on deposition onto the glass beads. The results of these experiments suggest that water treatment facilities treating source waters with moderate organic matter concentrations and/or employing biologically active filters have a greater potential for oocyst breakthrough and proper coagulation is critical for effective removal of oocysts in the filters.     

Ecological equilibrium on biological activated carbon

This paper examines the potential of a biological activity control system (BACS) for biological activated carbon (BAC) in comparison to granular activated carbon (GAC) for the treatment of potable water. The overall objective of the project is to produce drinking water of a higher quality more economically by developing a BACS for exhausted GAC that can be transformed to BAC by the development of a natural biofilm during the bio-regeneration mode. The research therefore may be interesting for water companies and the activated carbon industry. Findings show that the lifetime of a GAC filter can be significantly extended by maintaining an active biofilm that has to be controlled in order to avoid filter clogging. The most important parameters are dissolved oxygen (DO), pH and a correct balance of nutrients, which enables a natural control of the biomass. pH control was required to maintain an optimal bacteria-protozoa level. Excessive growth of filamentous bacteria can be prevented by a decrease in DO, increase in pH and the reduction of one essential nutrient, e.g. total phosphorus (P). Total organic carbon (TOC) and chemical oxygen demand (COD) values were reduced by bioactivity. DO, turbidity and suspended solids (SS) values were kept in acceptable ranges with respect to drinking water objectives. Plants without a significant population of protozoa deliver turbid low quality effluent high on SS and biochemical oxygen demand (BOD). It was possible to control the biofilm on GAC containing a natural biofilm and BAC during the bio-regeneration mode. Natural and artificial bio-regeneration lead to similar performance characteristics.     

Removal of MIB and geosmin using granular activated carbon with and without MIEX pre-treatment

This study assessed the impact of MIEX pre-treatment, followed by either coagulation or microfiltration (MF), on the effectiveness of pilot granular activated carbon (GAC) filters for the removal of the taste and odour compounds, 2-methylisoborneol (MIB) and geosmin, from a surface drinking water source over a 2-year period. Complete removal of MIB and geosmin was achieved by all GAC filters for the first 10 months, suggesting that the available adsorption capacity was sufficient to compensate for differences in dissolved organic carbon (DOC) entering the GAC filters.Reduction of empty bed contact time (EBCT), in all but one GAC filter, resulted in breakthrough of spiked MIB and geosmin, with initial results inconclusive regarding the impact of MIEX pre-treatment. MIB and geosmin removal increased over the ensuing 12 months until complete removal of both MIB and geosmin was again achieved in all but one GAC filter, which had been pre-chlorinated. Autoclaving and washing the GAC filters had minimal impact on geosmin removal but reduced MIB removal by 30% in all but the pre-chlorinated filter, confirming that biodegradation impacted MIB removal. The impact of biodegradation was greater than any impact on GAC adsorption arising from DOC differences due to MIEX pre-treatment. It is not clear whether, at a lower initial EBCT, MIEX pre-treatment may have impacted on the adsorption capacity of the virgin GAC.The GAC filter maintained at the longer EBCT, which was also pre-chlorinated, completely removed MIB and geosmin for the period of the study, suggesting that the greater adsorption capacity was compensating for any decrease in biological degradation.     

Impact of temperature on nitrification in biological activated carbon (BAC) filters used for drinking water treatment

The impact of temperature on nitrification in biological granular activated carbon (GAC) filters was evaluated in order to improve the understanding of the nitrification process in drinking water treatment. The study was conducted in a northern climate where very cold water temperatures (below 2 degrees C) prevail for extended periods and rapid shifts of temperature are frequent in the spring and fall. Ammonia removals were monitored and the fixed nitrifying biomass was measured using a method of potential nitrifying activity. The impact of temperature was evaluated on two different filter media: an opened superstructure wood-based activated carbon and a closed superstructure activated carbon-based on bituminous coal. The study was conducted at two levels: pilot scale (first-stage filters) and full-scale (second-stage filters) and the results indicate a strong temperature impact on nitrification activity. Ammonia removal capacities ranged from 40 to 90% in pilot filters, at temperatures above 10 degrees C, while more than 90% ammonia was removed in the full-scale filters for the same temperature range. At moderate temperatures (4-10 degrees C), the first stage pilot filters removed 10-40% of incoming ammonia for both media (opened and closed superstructure). In the full-scale filters, a difference between the two media in nitrification performances was observed at moderate temperatures: the ammonia removal rate in the opened superstructure support (more than 90%) was higher than in the closed superstructure support (45%). At low temperatures (below 4 degrees C) both media performed poorly. Ammonia removal capacities were below 30% in both pilot- and full-scale filters.     

Visualization of active biomass distribution in a BGAC fluidized bed reactor using GFP tagged Pseudomonas putida F1

A favorable microenvironment for biofilm growth on GAC particles was shown using green fluorescent protein (GFP) as a marker for a phenol degrading bacterium, Pseudomonas putida F1. The dispersion of P. putida F1 in a biofilm covering granulated activated carbon (GAC) particles was monitored and compared to a biofilm on non-activated granular carbon particles. Laser scanning confocal microscopy (LSCM) micrographs of the biofilms taken from two fluidized bed reactors operating under identical conditions, showed higher fluorescent green areas in the GAC biofilm, especially close to the GAC surface. Quantitative analysis of the biofilm by COMSTAT, a three-dimensional biofilm structure analysis program, showed higher biomass concentration and higher viability in the GAC covered biofilm vs. the non-activated carbon biofilm. In addition, better effluent quality was measured for the BGAC reactor, which strongly suggests a significantly larger biofilm surface area available to the substrate, as opposed to that of the non-activated carbon carrier reactor.     

The potential for biofilm growth in water distribution systems.

Biofilms on pipe walls in water distribution systems are composed of bacteria in a polymeric matrix, which can lead to chlorine demand, coliform growth, pipe corrosion and water taste and odour problems. The majority of previous studies have been laboratory or pilot plant based and few results are available for field conditions. In this study, field observations of biofilm were made using biofilm potential monitors. The monitor results were compared with pipe samples taken from the distribution system and with laboratory pipe reactors. An empirical equation quantified the inhibitory effects of free chlorine and decrease of temperature on biofilm growth. With water having total organic carbon concentrations in the range 1.5-3.9mg/1 a free chlorine residual of 0.2 mg/l was needed to reduce biofilm concentration to below 50 pg ATP cm2. Pipe material influenced biofilm activity far less than chlorine with mean biofilm activity being ranked in the order glass (136 pg ATP/cm2) < cement (212 pg ATP/cm2) < MDPE (302 pg ATP/ cm2) < PVC (509 pg ATP/cm2).     

DBPs removal in GAC filter-adsorber

A rapid sand filter and granular activated carbon filter-adsorber (GAC FA) were compared in terms of dissolved organic carbon (DOC) and disinfection by-products (DBPs) removal. A water treatment plant (WTP) that had a high ammonia concentration and DOC in raw water, which, in turn, led to a high concentration of DBPs because of a high dose of pre-chlorination, was investigated. To remove DBPs and DOC simultaneously, a conventional rapid sand filter had been retrofitted to a GAC FA at the Buyeo WTP in Korea. The overall removal efficiency of DBPs and DOC was higher in the GAC FA than in the sand filter, as expected. Breakthrough of trihalomethanes (THMs) was noticed after 3 months of GAC FA operation, and then removal of THMs was minimal (<10%). On the other hand, the removal efficiency of five haloacetic acids (HAA(5)) in the GAC FA was better than that of THMs, though adsorption of HAA(5) decreased rapidly after 3.5 months of GAC FA operation. And then, gradual improvement (>90%) in HAA(5) removal efficiency was again observed, which could be attributed to biodegradation. At the early stage of GAC FA operation, HAA(5) removal was largely due to physical adsorption, but later on biodegradation appeared to prevail. Biodegradation of HAA(5) was significantly influenced by water temperature. Similar turbidity removal was noticed in both filters, while better manganese removal was confirmed in the sand filter rather than in the GAC FA.     

Experimental results and mathematical modelling of an autotrophic and heterotrophic biofilm in a sand filter treating landfill leachate and municipal wastewater

A better understanding of wastewater treatment with soil filters is important to optimise plant operation and reduce the risk of clogging. The article presents results of a treatment concept which uses a combination of SBR and vertical-flow sand filter technology. The SBR was mainly used for denitrification and sedimentation of substances in particulate form. Efficient nitrification was achieved by the planted sand filter. Degradation rates of 10gNH(4)-N/(m(2)xd) were measured for periods with peak loadings. The two-dimensional dynamic model reproduces the biofilm growth and decay of heterotrophic and autotrophic biomass. It is capable of describing the clogging of the sand filter by combining a biochemical and a geometric model. After calibration, the model was used for the calculation of maximum nitrogen degradation performances. Maximum degradation rates of 12gNH(4)-N/(m(2)xd) can be achieved if the COD/TKN ratio is reduced before to a level lower than that of municipal wastewater. The COD was further degraded in the filter than we expected comparing it with activated sludge plants. Within the soil filter a biofilm thickness of up to 110mum is simulated depending on the embankment of gravel and grains of sand. Sensitivity analysis of model parameters showed the high impact of the maximum autotrophic growth rate, the autotrophic yield, the diffusion coefficient for oxygen and the number of contact points of the single grains of sand.     

生物砂滤池不同挂膜方法的试验研究

根据生物砂滤池的运行特点，通过试验对自然挂膜和接种挂膜两种不同挂膜方法下，各滤柱中的生物量和生物活性，对CODMn、NH4^＋ -N和UV254的去除效果以及运行周期进行了分析研究。结果表明，接种挂膜的启动时间比自然挂膜缩短了6～8d，同时生物量较多，但生物活性比自然挂膜法低12．03％～14．29％；各柱对CODMn、NH4^＋ -N均有很好的去除效果，平均去除率在35％和80％以上；但采用自然挂膜法对污染物的去除效果更佳，特别是对UV254的去除率较接种挂膜法提高了5．01％～10．8％，并且运行周期延长了6～10h，运行更为稳定。