Association between phosphorus and suspended solids in an Everglades treatment wetland dominated by submersed aquatic vegetation

Restoration of the Everglades requires reduction of total phosphorus (TP) in the influent run-off from the Everglades agricultural area (EAA). The Everglades nutrient removal project tested phosphorus (P) - removal efficiencies of several treatment wetland cells. The best TP reduction has occurred within the submersed aquatic vegetation (SAV) - dominated treatment Cell 4. A significant proportion of the P reduction in Cell 4 over several years has been in the form of particulate P (PP). This study was conducted to (i) determine and compare the components of suspended solids in the Cell 4 influent and effluent waters, and (ii) investigate associations between PP and individual particulate components. Identification and quantification of components were accomplished using X-ray diffraction, thermogravimetry, scanning electron microscopy, and energy dispersive X-ray elemental analysis. The dominant particulate components in the Cell 4 water column are organic matter (OM), biogenic Si (predominantly diatom frustules), and calcite. Concentrations of PP, suspended solids, and particulate OM were greater at the Cell 4 inflow than at the outflow; consistent differences between particulate calcite in the influent vs. the effluent were not found. PP was positively correlated with particulate OM, but was not correlated with calcite. Data suggest that particulate OM, including microbial cells, plays an important role in P transport from the EAA. Possibly, a shift from planktonic to periphytic microbial distribution contributes to PP reduction. The importance of planktonic organisms as vectors of P in Everglades water warrants further study.     

A Review of Processes Responsible for Metal Removal in Wetlands Treating Contaminated Mine Drainage

Many reports have documented wetlands removing a wide variety of contaminants in mine drainage, including aluminum, arsenic, cadmium, cobalt, copper, cyanide, iron, lead, manganese, nickel, selenium, uranium, and zinc. This article reviews biogeochemical processes responsible for their ability to transform and retain metals into insoluble forms. Shallow depth and large inputs of organic matter are key characteristics of wetlands that promote chemical and biological processes effecting metal removal. Aquatic macrophytes play an essential role in creating and maintaining this environment, but their uptake of metals usually accounts for a minor proportion of the total mass removed. Sorption onto organic matter is important in metal removal, particularly for copper, nickel, and uranium. Aluminum, iron, and manganese are often removed by hydrolysis, with the resulting acidification of water buffered by alkalinity produced in wetland sediments by anaerobic bacteria. Bacterial sulfate reduction accounts for much of this alkalinity. It can also contribute significantly to metal removal by formation of insoluble sulfides. Other important processes include the formation of insoluble carbonates, reduction to nonmobile forms, and adsorption onto iron oxides and hydroxides. Examples from field studies are presented throughout the review to illustrate these processes.     

Phytostabilization of gold mine tailings, New Zealand. Part 1: Plant establishment in alkaline saline substrate

Tailings from the Macraes mine, southern New Zealand, are prone to wind erosion. Use of a vegetation cover for physical stabilization is one potential solution to this environmental problem. This study used field trials contained in lysimeters to 1), test the ability of different plant species to grow in un/amended tailings and 2), provide background information on the nutrient and chemical content of waters in tailings. Barley (Hordeum vulgare), blue lupin (Lupinus angustifolius), and rye corn (Secale cereale) were trialed, using Superphosphate fertilizer and sewage sludge as amendments. Rye corn grew well in fertilizer-amended tailings, but poorly in unamended tailings; barley growth was similar in amended and unamended tailings; blue lupins grew poorly overall The tailings had alkaline pH (7-8.5) and water rapidly (< 1 mo) interacted with the tailings to become strongly saline. Minor acid generation was neutralized by calcite, with associated release of calcium and carbonate ions. Leachate waters were supersaturated with respect to calcite and dolomite. Dissolved sodium concentrations were up to 1000 mg L(-1), but elevated Ca2+ calcium and Mg2+ ensured that sodicity was lower than plant-toxic levels. Rye corn is a potentially useful plant for rapid phytostabilization of tailings, with only minor phosphate amendment required.     

Effect of mineral composition on bacterial attachment to submerged rock surfaces

A direct microscopic count technique employing fluorescein isothiocyanate stain was used to compare microbial colonization on the exposed surfaces of rocks and minerals suspended in several ponds for various time intervals. Hematitic sandstone was never colonized at a rate greater than limestone, but quartz was always colonized more rapidly than calcite. The use of single-crystal minerals (quartz and calcite) in a nested factor experiment showed that the effect of the minerals on colonization was statistically significant, but that differences among the immersion sites were also significant. Sandstone samples placed in a pond outflow accumulated microbial colonizers more rapidly than those placed in the still waters of the same pond. The results indicate that the composition of the mineral substrate, in concert with the immersion environment, controls the formation of primary slime layers in aquatic systems.     

The influence of environmental factors on the rate and extent of stainless steel ennoblement mediated by manganese‐oxidizing biofilms

The increase in the open circuit potential of passive metals in natural waters due to biofilm formation at the metal surface, termed ennoblement, has been reported for nearly 30 years. Although its occurrence is undoubtedly associated with microbial colonization, the underlying mechanism of ennoblement remains controversial. Recent work produced in the authors’ laboratory has provided convincing experimental evidence that ennoblement can be caused by deposition of biomineralized manganese produced by manganese‐oxidizing biofilms. The purpose of this study was to determine the effects of environmental factors on the rate and extent of ennoblement of 316L stainless steel exposed to natural waters. This was accomplished by exposing corrosion coupons to four freshwater systems over a 4‐yearperiod. The rate and extent of ennoblement observed in these locations was correlated with dissolved manganese concentrations, the mass of accumulated manganese oxides, organic carbon concentration, dissolved oxygen concentration, flow, conditions, temperature, and pH in these environments.     

Geochemistry and microbiology of an impounded subterranean acidic water body at Mynydd Parys, Anglesey, Wales

Acidic, metal‐rich water that had accumulated within two abandoned, adjacent copper mines in north Wales was removed to prevent its possible catastrophic release. About 274 000 m³ of acidic (pH ～2.4) mine water was pumped out of the mines over a 14‐week period. Concentrations of dissolved species (iron, sulfate, aluminium, copper, manganese and zinc) increased as water at lower depths within the mines was accessed. The discharged water flowed through a small wetland area, reaching the sea about 3 km north of the mine site. Analysis of the water at three sampling stations revealed that there was very little removal of aluminium and most of the heavy metals present except (ferrous) iron, which was partially removed as a result of oxidation and hydrolysis of the resulting ferric iron. The dominant bacterium in the subterranean mine water, which was essentially devoid of oxygen, was the iron‐ and sulfur‐oxidizer Acidithiobacillus ferrooxidans. The microbial populations in the pumped mine water were monitored using combined cultivation‐dependent (isolation on solid media) and cultivation‐independent (terminal restriction fragment length polymorphism and clone library) techniques. Other bacteria detected in the mine water included other iron‐oxidizers (Leptospirillum spp., ‘Ferrimicrobium acidiphilum’ and Gallionella‐like organisms) and heterotrophic acidophiles (Acidiphilium, Acidisphaera and Acidobacterium). Archaeal clones were also detected; most of these were related to methanogens. Owing to the absence of an effective remediation strategy, an estimated 7.5 tonnes of copper, 3.1 tonnes of manganese, 14.8 tonnes of zinc and 15.3 tonnes of aluminium was discharged into the Irish Sea as a consequence of the dewatering of the mines.     

Microbial communities and geochemical dynamics in an extremely acidic, metal-rich stream at an abandoned sulfide mine (Huelva, Spain) underpinned by two functional primary production systems

An extremely acidic (pH 2.5-2.75) metal-rich stream draining an abandoned mine in the Iberian Pyrite Belt, Spain, was ramified with stratified macroscopic gelatinous microbial growths ('acid streamers' or 'mats'). Microbial communities of streamer/mat growths sampled at different depths, as well as those present in the stream water itself, were analysed using a combined biomolecular and cultivation-based approach. The oxygen-depleted mine water was dominated by the chemolithotrophic facultative anaerobe Acidithiobacillus ferrooxidans, while the streamer communities were found to be highly heterogeneous and very different to superficially similar growths reported in other extremely acidic environments. Microalgae accounted for a significant proportion of surface streamer biomass, while subsurface layers were dominated by heterotrophic acidophilic bacteria (Acidobacteriacae and Acidiphilium spp.). Sulfidogenic bacteria were isolated from the lowest depth streamer growths, where there was also evidence for selective biomineralization of copper sulfide. Archaeal clones (exclusively Euryarchaeota) were recovered from streamer samples, as well as the mine stream water. Both sunlight and reduced inorganic chemicals (predominantly ferrous iron) served as energy sources for primary producers in this ecosystem, promoting complex microbial interactions involving transfer of electron donors and acceptors and of organic carbon, between microorganisms in the stream water and the gelatinous streamer growths. Microbial transformations were shown to impact the biogeochemical cycling of iron and sulfur in the acidic stream, severely restricting the net oxidation of ferrous iron even when the initially anoxic waters were oxygenated by indigenous acidophilic algae. A model accounting for the biogeochemistry of iron and sulfur in the mine waters is described, and the significance of the acidophilic communities in regulating the geochemistry of acidic, metal-rich waters is described.     

A process-orientated design and performance assessment methodology for passive mine water treatment systems

The aim of this paper is the development of a methodology for assessing the iron removal efficiency of passive mine water treatment settling lagoons and reed beds. Previous work in the design and sizing of coal mine drainage lagoons and wetlands has focussed on the use of standard hydraulic residence times or using the 10 g m⁻² d⁻¹ metric, these criteria have been applied without regard to the fundamental physical and chemical processes controlling iron removal in the system, namely the pH dependent rates of Fe(II) oxidation and physical settling of the particulate Fe(III). In this study field water quality data have been collected from lagoons and reed beds. These data are examined alongside data from the UK Coal Authority database and combined with simple mathematical formulations to provide a framework through which to understand passive treatment schemes from a process-orientated perspective. It is demonstrated that for the sites studied reed beds are more efficient for iron removal. This paper recommends that judgements of iron removal performance should be based on a derived treatment efficiency index (?) and that the modelling approach espoused in this paper should be used when designing passive mine water treatment schemes.     

Selective removal of transition metals from acidic mine waters by novel consortia of acidophilic sulfidogenic bacteria

Two continuous‐flow bench‐scale bioreactor systems populated by mixed communities of acidophilic sulfate‐reducing bacteria were constructed and tested for their abilities to promote the selective precipitation of transition metals (as sulfides) present in synthetic mine waters, using glycerol as electron donor. The objective with the first system (selective precipitation of copper from acidic mine water containing a variety of soluble metals) was achieved by maintaining a bioreactor pH of ～2.2–2.5. The second system was fed with acidic (pH 2.5) synthetic mine water containing 3 mM of both zinc and ferrous iron, and varying concentrations (0.5–30 mM) of aluminium. Selective precipitation of zinc sulfide was possible by operating the bioreactor at pH 4.0 and supplementing the synthetic mine water with 4 mM glycerol. Analysis of the microbial populations in the bioreactors showed that they changed with varying operational parameters, and novel acidophilic bacteria (including one sulfidogen) were isolated from the bioreactors. The acidophilic sulfidogenic bioreactors provided ‘proof of principle’ that segregation of metals present in mine waters is possible using simple online systems within which controlled pH conditions are maintained. The modular units are versatile and robust, and involve minimum engineering complexity.     

Abundant and diverse arsenic-metabolizing microorganisms in peatlands treating arsenic-contaminated mining wastewaters

Mining operations produce large quantities of wastewater. At a mine site in Northern Finland, two natural peatlands are used for the treatment of mining-influenced waters with high concentrations of sulphate and potentially toxic arsenic (As). In the present study, As removal and the involved microbial processes in those treatment peatlands (TPs) were assessed. Arsenic-metabolizing microorganisms were abundant in peat soil from both TPs (up to 10⁸ cells g_dw⁻¹), with arsenate respirers being about 100 times more abundant than arsenite oxidizers. In uninhibited microcosm incubations, supplemented arsenite was oxidized under oxic conditions and supplemented arsenate was reduced under anoxic conditions, while little to no oxidation/reduction was observed in NaN₃-inhibited microcosms, indicating high As-turnover potential of peat microbes. Formation of thioarsenates was observed in anoxic microcosms. Sequencing of the functional genemarkers aioA (arsenite oxidizers), arrA (arsenate respirers) and arsC (detoxifying arsenate reducers) demonstrated high diversity of the As-metabolizing microbial community. The microbial community composition differed between the two TPs, which may have affected As removal efficiencies. In the present situation, arsenate reduction is likely the dominant net process and contributes substantially to As removal. Changes in TP usage (e.g. mine closure) with lowered water tables and heightened oxygen availability in peat might lead to re-oxidation and re-mobilization of bound arsenite.     

Marble stone processing powder residue as chemical adjuvant for the biologic treatment of acid mine drainage

Marble stone powder (calcite tailing) is a residual material resulting from the cutting and polishing of marble stone. Its use as adjuvant for the biologic treatment of acid mine drainage is studied. The performance of a combined chemical/biologic packed bed reactor containing a sulphate-reducing bacteria inoculum and calcite tailing as neutralising agent was compared with the individual neutralisation and biologic steps. Unlike the individual steps, the combined column is efficient in terms of metals and sulphate removal and acidity neutralisation, producing treated water suitable for irrigation. This work emphasizes on the key role played by waste calcite tailing in the inlet of the reactor. By thus adjusting pH to values adequate for sulphate-reducing bacteria water suitable for irrigation can be obtained in a single equipment with low energy demand. This goal would be impossible without the neutralisation action.     

Microbial Manganese Oxidation in the Lower Mississippi River: Methods and Evidence

Abstract

Recent work has led to the suggestion that biologically-mediated redox processes might be important in the regulation of dissolved trace element concentrations in rivers, especially with regard to manganese. Here, we focus on the removal of dissolved Mn from lower Mississippi River water. Experiments indicate that dissolved Mn can be rapidly removed from lower Mississippi River water on a timescale of days or less and that Mn oxides are formed. However, demonstrating a biological origin for this removal is problematic. Experiments reveal that commonly used microbial controls, including NaN₃ mixtures, HgCl₂, heat sterilization, and sonification all affect fluvial particulate Mn through dissolution, disaggregation, interference with adsorption, or particle ageing. Thus, these microbial controls may affect abiotic as well as biological processes. Evidence supporting microbial removal of dissolved Mn from lower Mississippi River water includes a temperature optimum for the process (～30°C), a lower activation energy than reported for heterogeneous inorganic Mn oxidation, and a faster rate than reported for autocatalytic inorganic Mn oxidation. This rapid Mn oxidation process occurs at essentially the same rate in the dark as well as the light. Observation of Mn removal at similar rates in a blackwater river in addition to the lower Mississippi, suggests that this is a common phenomenon in fluvial systems. If, as has been shown by other lab studies, the freshly biologically precipitated Mn oxides have a high specific surface area, then our observations provide a potential link between microbial activity, Mn cycling, and the cycling of other particle-reactive trace elements in rivers. Our results also indicate that unfiltered river water samples for dissolved Mn analysis should be filtered as soon as possible or at least stored cold if immediate filtration is not possible.     

紫金山铜矿酸性矿山废水微生物群落多样性

【背景】为避免环境污染,酸性矿山废水需经处理后才能排放,处理后的废水理化性质会发生显著变化,将影响整个微生物群落的结构。【目的】分析处理前后的细菌和真菌群落变化及其与理化参数的关系,为矿山废水的处理提供参考指标,并为矿山污染场地的修复提供理论基础。【方法】采集福建紫金山铜矿的酸性矿山废水并测定其理化性质。采用基于原核微生物16S rRNA基因V4区和真菌18S rRNA基因ITS的高通量测序技术分析水样的微生物群落结构。【结果】经中和处理后的回水与矿坑水和生物浸出液相比,pH升高,重金属离子含量显著降低。原核微生物的多样性高于真菌,回水的物种多样性高于矿坑水和浸出液。回水中变形菌门的丰度最高,矿坑水和浸出液中分别以广古菌门和硝化螺菌门的丰度最高。回水中噬氢菌属为优势类群,矿坑水和浸出液中的优势菌是钩端螺旋菌属,铁质菌属等古菌也有一定的比例。pH、Al、Mn、Zn与回水中相对丰度较高的菌属显著相关,而矿坑水和浸出液中的高丰度类群与环境因子没有显著的相关性。【结论】研究表明酸性废水的中和沉淀处理对微生物群落产生了较大的影响,微生物群落变化可以作为矿山酸性废水污染处理效果的一个参考指标。     

Hydrogeochemistry of Lake Gallocanta (Aragón,NE Spain)

Lake Gallocanta has undergone drastic changes during the last thirteen years. Water level changed from a high level (Z_max = 2 m) to total dryness in 1985. From 1986 to 1988 slow refilling occurred. The water volume fluctuations have been studied in relation to climatic variations recorded for that period. Variations in the major dissolved ions were related to water volume fluctuations from data at two different stages, one corresponding to the drying phase and another to the refilling phase. Mineralogical composition of the salts precipitated at different stages was examined by X-ray diffraction. Interstitial water and mineralogical composition of recent sediments were also studied along a transect through the lake.The water column decrease from 1977 to 1985 is related to decreasing annual rainfall (500-250 mm respectively). The refilling in 1986–1988 is due to high annual rainfall (537 mm). In addition to these fluctuations, seasonal changes of the water level between 20 and 50 cm occurred every year.Gallocanta is a Na-Mg-Cl-(SO₄) type lake. During the drying period a typical salt enrichment occurs with linear relationships between TDS, Cl, Na and K. Alkalinity is linearly correlated with Ca at relatively low salinities. As salinity increases a linear relationship between Ca and SO₄ is observed. Minerals formed from the brine are halite, bischofite, epsomite, hexahydrite, mirabilite, gypsum, aragonite, calcite and dolomite. The molar ratio Mg/Ca of the interstitial water changes from 1.5 along the shorelines, where calcite and aragonite precipitate, to 40 in the center of the lake. Sediment cores from the central part of the lake show aragonite in the top layers, magnesian calcite and low proportions of quartz and illite, while at 20 cm depth a high proportion of gypsum is present. In contrast, cores from the shore of the lake are mainly composed of low magnesium calcite in the top layers and low magnesium calcite together high magnesium calcite and dolomite between 30 and 70 cm depth. Gypsum deposits only occur in significant proportions at 80–100 cm depth.The refilling process showed relationships between volume and salt concentration following the Langbein model. The salt mass in solution decreased about 50% from the drying to the refilling phase. However, Mg content decreased about 70 % for the same period, suggesting a contribution of this element to the dolomite formation.     

Deposition of manganese in a drinking water distribution system

The deposition of manganese in a water distribution system with manganese-related "dirty water" problems was studied over a 1-year period. Four monitoring laboratories with Robbins biofilm sampling devices fitted to the water mains were used to correlate the relationship among manganese deposition, the level of manganese in the water, and the chlorination conditions. Manganese deposition occurred by both chemical and microbial processes. Chemical deposition occurred when Mn(II) not removed during water treatment penetrated the filters and entered the distribution system, where it was oxidized by chlorine and chlorine dioxide used for disinfection. Microbial deposition occurred in areas with insufficient chlorination to control the growth of manganese-depositing biofilm. At 0.05 mg of Mn(II) per liter, the chemical deposition rate was much greater than microbial deposition. Significant deposition occurred at 0.03 mg of manganese per liter, and dirty water complaints were not eliminated until manganese levels were continuously less than 0.02 mg/liter and chlorination levels were greater than 0.2 mg/liter. A guideline level of 0.01 mg of manganese per liter is recommended.     

Deposition of manganese in a drinking water distribution system.

The deposition of manganese in a water distribution system with manganese-related "dirty water" problems was studied over a 1-year period. Four monitoring laboratories with Robbins biofilm sampling devices fitted to the water mains were used to correlate the relationship among manganese deposition, the level of manganese in the water, and the chlorination conditions. Manganese deposition occurred by both chemical and microbial processes. Chemical deposition occurred when Mn(II) not removed during water treatment penetrated the filters and entered the distribution system, where it was oxidized by chlorine and chlorine dioxide used for disinfection. Microbial deposition occurred in areas with insufficient chlorination to control the growth of manganese-depositing biofilm. At 0.05 mg of Mn(II) per liter, the chemical deposition rate was much greater than microbial deposition. Significant deposition occurred at 0.03 mg of manganese per liter, and dirty water complaints were not eliminated until manganese levels were continuously less than 0.02 mg/liter and chlorination levels were greater than 0.2 mg/liter. A guideline level of 0.01 mg of manganese per liter is recommended.     

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.     

Archaeal diversity in waters from deep South African gold mines.

A culture-independent molecular analysis of archaeal communities in waters collected from deep South African gold mines was performed by performing a PCR-mediated terminal restriction fragment length polymorphism (T-RFLP) analysis of rRNA genes (rDNA) in conjunction with a sequencing analysis of archaeal rDNA clone libraries. The water samples used represented various environments, including deep fissure water, mine service water, and water from an overlying dolomite aquifer. T-RFLP analysis revealed that the ribotype distribution of archaea varied with the source of water. The archaeal communities in the deep gold mine environments exhibited great phylogenetic diversity; the majority of the members were most closely related to uncultivated species. Some archaeal rDNA clones obtained from mine service water and dolomite aquifer water samples were most closely related to environmental rDNA clones from surface soil (soil clones) and marine environments (marine group I [MGI]). Other clones exhibited intermediate phylogenetic affiliation between soil clones and MGI in the Crenarchaeota. Fissure water samples, derived from active or dormant geothermal environments, yielded archaeal sequences that exhibited novel phylogeny, including a novel lineage of Euryarchaeota. These results suggest that deep South African gold mines harbor novel archaeal communities distinct from those observed in other environments. Based on the phylogenetic analysis of archaeal strains and rDNA clones, including the newly discovered archaeal rDNA clones, the evolutionary relationship and the phylogenetic organization of the domain Archaea are reevaluated.     

Interaction of mineral elements in sea water and shell of oysters (Crassostrea virginica (Gmelin)) cultured in controlled and natural systems

The interaction is reported of selected chemical elements (cadmium, calcium, copper, iron, magnesium, manganese, strontium, and zinc) in cultured sea water, with soft tissues, prismatic calcite of the right valve, and foliated calcite of right and left valves of genetically similar American oysters, Crassostrea virginica (Gmelin) grown in a natural habitat and in two environmentally controlled experimental systems (flow-through and recycle). The addition of trace elements as algal nutrients in ambient sea water was reflected in higher concentrations of trace metals in shells and soft tissues of oysters grown in experimental systems. Calcium was relatively uniformly distributed in major regions of valves from the three habitats, even though its concentration fluctuated widely in sea water in experimental systems. Magnesium and strontium were most concentrated in valves of oysters grown in the recycle system (magnesium in the prismatic layer of the shell and strontium in the foliated calcite). Iron was uniformly distributed. Cadmium, copper, manganese, and zinc were most concentrated in the prismatic calcite of valves from the flow-through system. In soft tissues, calcium was more concentrated in oysters from experimental systems than in those from the natural habitat. Manganese was about equally distributed in soft tissues from the three habitats, whereas copper and iron were more concentrated in soft tissues in experimental systems than in the natural habitat, and were many times more concentrated in soft tissues than in valves from all three habitats. As concentrations of magnesium, strontium, mangenese, zinc, and cadmium increased in valves in experimental systems, pigmentation of valves decreased. The study confirmed the capacity of oysters to concentrate several elements in their valves as concentration of these elements increased in ambient sea water and disclosed the heterogeneous distribution of these elements in major regions of the valves.     

Bacterial colonization of pellet softening reactors used during drinking water treatment

Pellet softening reactors are used in centralized and decentralized drinking water treatment plants for the removal of calcium (hardness) through chemically induced precipitation of calcite. This is accomplished in fluidized pellet reactors, where a strong base is added to the influent to increase the pH and facilitate the process of precipitation on an added seeding material. Here we describe for the first time the opportunistic bacterial colonization of the calcite pellets in a full-scale pellet softening reactor and the functional contribution of these colonizing bacteria to the overall drinking water treatment process. ATP analysis, advanced microscopy, and community fingerprinting with denaturing gradient gel electrophoretic (DGGE) analysis were used to characterize the biomass on the pellets, while assimilable organic carbon (AOC), dissolved organic carbon, and flow cytometric analysis were used to characterize the impact of the biological processes on drinking water quality. The data revealed pellet colonization at concentrations in excess of 500 ng of ATP/g of pellet and reactor biomass concentrations as high as 220 mg of ATP/m(3) of reactor, comprising a wide variety of different microorganisms. These organisms removed as much as 60% of AOC from the water during treatment, thus contributing toward the biological stabilization of the drinking water. Notably, only a small fraction (about 60,000 cells/ml) of the bacteria in the reactors was released into the effluent under normal conditions, while the majority of the bacteria colonizing the pellets were captured in the calcite structures of the pellets and were removed as a reusable product.