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Previous studies have shown that membrane-aerated biofilm (MAB) reactors can simultaneously remove carbonaceous and nitrogenous pollutants from wastewater in a single reactor. Oxygen is provided to MABs through gas-permeable membranes such that the region nearest the membrane is rich in oxygen but low in organic carbon, whereas the outer region of the biofilm is void of oxygen but rich in organic carbon. In this study, MABs were grown under similar conditions but at two different fluid velocities (2 and 14 cm s ؊1 ) across the biofilm. MABs were analyzed for changes in biomass density, respiratory activity, and bacterial community structure as functions of biofilm depth. Biomass density was generally highest near the membrane and declined with distance from the membrane. Respiratory activity exhibited a hump-shaped profile, with the highest activity occurring in the middle of the biofilm. Community analysis by PCR cloning and PCR-denaturing gradient gel electrophoresis of 16S rRNA genes demonstrated substantial stratification of the community structure across the biofilm. Population profiles were also generated by competitive quantitative PCR of gene fragments specific for ammonia-oxidizing bacteria (AOB) (amoA) and denitrifying bacteria (nirK and nirS). At a flow velocity of 14 cm s ؊1 , AOB were found only near the membrane, whereas denitrifying bacteria proliferated in the anoxic outer regions of the biofilm. In contrast, at a flow velocity of 2 cm s ؊1 , AOB were either not detected or detected at a concentration near the detection limit. This study suggests that, under the appropriate conditions, both AOB and denitrifying bacteria can coexist within an MAB.The aeration of bioreactors is one of the most significant operating costs incurred during the treatment of municipal and industrial wastewaters. One technological alternative to conventional coarse and fine bubble diffusers is the use of microporous membranes that contain small (Ͻ0.1 m) hydrophobic pores for bubbleless oxygen transfer to the wastewater (1). Biofilms actively grow on these membranes, such that membrane-aerated biofilm (MAB) reactors can treat wastewater at rates comparable to conventional suspended-growth processes (6,33,37). MAB reactors have lower operating costs, because oxygen transfer efficiency approaches 100% (5, 26) and because expensive air compression is no longer required (9). In addition, MAB reactors generate fewer odors, because volatile pollutants are not released into the atmosphere (5, 6).MABs are unique compared to biofilms grown on inert substrata. Nutrient concentrations in MABs are highest at the biofilm-liquid boundary and decrease with depth into the biofilm; in contrast, oxygen concentrations are highest at the membrane and decrease toward the outside of the biofilm (Fig. 1). In thick MABs, oxygen partially penetrates the biofilm, generating an anoxic zone at the biofilm-liquid boundary (6). In such biofilms, the region of greatest microbial activity is variable and depends on the relative availability of oxygen a...

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It is important to control biofilm cohesiveness to optimize process performance. In this study, a membraneaerated biofilm reactor inoculated with activated sludge was used to grow mixed-culture biofilms of different ages and thicknesses. The cohesions, or cohesive energy levels per unit volume of biofilm, based on a reproducible method using atomic force microscopy (F. Ahimou, M. J. Semmens, P. J. Novak, and G. Haugstad, Appl. Environ. Microbiol. 73:2897-2904, 2007), were determined at different locations within the depths of the biofilms. In addition, the protein and polysaccharide concentrations within the biofilm depths, as well as the dissolved oxygen (DO) concentration profiles within the biofilms, were measured. It was found that biofilm cohesion increased with depth but not with age. Level of biofilm cohesive energy per unit volume was strongly correlated with biofilm polysaccharide concentration, which increased with depth in the membrane-aerated biofilm. In a 12-day-old biofilm, DO also increased with depth and may therefore be linked to polysaccharide production. In contrast, protein concentration was relatively constant within the biofilm and did not appear to influence cohesion.Biofilms are ubiquitous in nature, and they can be beneficial or troublesome, depending upon where and how they grow. There appears to be a consensus that the content of extracellular polymeric substances (EPS) is important in biofilm cohesion and biofilm adhesion to surfaces. For example, Klapper et al. (17) used a model based on polymer viscoelastic properties and suggested that the material properties of biofilm were largely determined by the EPS, implying that biofilm strength should indeed be linked to EPS quantity and composition. In addition, a recent study by Xavier et al. (35) proposed a kinetic model to describe biofilm detachment that was based on enzymatic disruption of the EPS matrix, thereby affecting biofilm cohesiveness.The EPS content of a biofilm can differ in quantity and character as a result of environmental factors. Numerous environmental factors have been reported to promote EPS production. These include high levels of oxygen (4), limited availability of nitrogen (15,22), desiccation (25), low temperature (16), low pH (28), and nutrient deprivation (20). Weiner et al. (34) described several roles and functions for EPS, including that of protection against environmental stress. In addition, Davies et al. (8) showed that activation of a gene (algC) for production of the exopolymer alginate was higher for Pseudomonas aeruginosa when attached to a Teflon mesh than for unattached P. aeruginosa. This suggests that organisms are able to respond to their environments and change EPS compositions and therefore their adhesion abilities, based on the surfaces to which they attach. Multivalent cations, such as those of calcium and magnesium, also probably play a role in the cohesiveness of microbial aggregates, as evaluated from studies of anaerobic sludge granules (12), activated sludge flocs (13), and biofilms ...