Biofilm Community Structure Research Articles

Microbial diversity, ecological networks and functional traits associated to materials used in drinking water distribution systems

Drinking water distribution systems host complex microbial communities as biofilms that interact continuously with delivered water. Understanding the diversity, behavioural and functional characteristics will be a requisite for developing future monitoring strategies and protection against water-borne health risks. To improve understanding, this study investigates mobilisation and accumulation behaviour, microbial community structure and functional variations of biofilms developing on different pipe materials from within an operational network. Samples were collected from four pipes during a repeated flushing operation three months after an initial visit that used hydraulic forces to mobilise regenerating biofilms yet without impacting the upstream network. To minimise confounding factors, test sections were chosen with comparable daily hydraulic regimes, physical dimensions, and all connected straight of a common trunk main and within close proximity, hence similar water chemistry, pressure and age. Taxonomical results showed differences in colonising communities between pipe materials, with several genera, including the bacteria Pseudomonas and the fungi Cladosporium, present in every sample. Diverse bacterial communities dominated compared to more homogeneous fungal, or mycobiome, community distribution. The analysis of bacterial/fungal networks based on relative abundance of operational taxonomic units (OTUs) indicated microbial communities from cast iron pipes were more stable than communities from the non-ferrous pipe materials. Novel analysis of functional traits between all samples were found to be mainly associated to mobile genetic elements that play roles in determining links between cells, including phages, prophages, transposable elements, and plasmids. The use of functional traits can be considered for development in future surveillance methods, capable of delivering network condition information beyond that of limited conventional faecal indicator tests, that will help protect water quality and public health.

Three-dimensional carbon-based anodes promoted the accumulation of exoelectrogens in bioelectrochemical systems.

To achieve deep understandings on the effects of structure and surface properties of anode material on the performance of bioelectrochemical systems, the present research investigated the bacterial community structures of biofilms attached to different three-dimensional anodes including carbon felt and materials derived from pomelo peel, kenaf stem, and cardboard with 454 pyrosequencing analysis based on the bacterial 16S rRNA gene. The results showed that bacterial community structures, especially the relative abundance of exoelectrogens, were significantly related to the types of adopted three-dimensional anode materials. Proteobacteria was the shared predominant phylum, accounting for 55.4%, 52.1%, 66.7%, and 56.1% for carbon felt, cardboard, pomelo peel, and kenaf stem carbon, respectively. The most abundant OTU was phylogenetically related to the well-known exoelectrogen of Geobacter, with a relative abundance of 16.3%, 19.0%, 36.3%, and 28.6% in carbon felt, cardboard, pomelo peel, and kenaf stem, respectively. Moreover, another exoelectrogen of Pseudomonas sp. accounted for 4.9% in kenaf stem and 3.9% in carbonboard, respectively. The results implied the macrostructure and properties of different anode materials might result in different niches such as hydrodynamics and substrate transport dynamics, leading to different bacterial structure, especially different relative abundance of exoelectrogens, which consequently affected the performance of bioelectrochemical systems. PRACTITIONER POINTS: Bioelectrochemical systems (BESs) represent a novel biotechnology platform to simultaneously treat wastewaters and produce electrical power. Three-dimensional materials derived from nature plant as anode to promote electricity output from BESs and reduce the construct cost of BESs. Macrostructure of the three-dimensional anode material affected phylotype richness and phylogenetic diversity of microorganisms in anodic biofilm of BESs. Geobacter as well-known exoelectrogen was the most abundant in biofilm attached to three-dimensional anode.

Changes in Microbial Community Structures under Reclaimed Water Replenishment Conditions

Using reclaimed water as a resource for landscape water replenishment may alleviate the major problems of water resource shortages and water environment pollution. However, the safety of the reclaimed water and the risk of eutrophication caused by the reclaimed water replenishment are unclear to the public and to the research community. This study aimed to reveal the differences between natural water and reclaimed water and to discuss the rationality of reclaimed water replenishment from the perspective of microorganisms. The microbial community structures in natural water, reclaimed water and natural biofilms were analyzed, and the community succession was clarified along the ecological niches, water resources, fluidity and time using 16S rRNA gene amplicon sequencing. Primary biofilms without the original community were added to study the formation of microbial community structures under reclaimed water acclimation. The results showed that the difference caused by ecological niches was more than those caused by the fluidity of water and different water resources. No significant difference caused by the addition of reclaimed water was found in the microbial diversity and community structure. Based on the results of microbial analysis, reclaimed water replenishment is a feasible solution that can be used for supplying river water.

The response of nitrogen cycling and bacterial communities to E. coli invasion in aquatic environments with submerged vegetation

The effects of exogenous Escherichia coli on nitrogen cycling (N-cycling) in freshwater remains unclear. Thus, seven ecosystems, six with submerged plants-Potamogeton crispus (PC) and Myriophyllum aquaticum (MA)-and one with no plants were set up. Habitats were assessed before and after E. coli addition (107colony-forming units/mL). E. coli colonization of freshwater ecosystems had significant effects on bacterial community structure in plant surface biofilms and surface sediments (ANOVA, P<0.05). It reduced the relative abundance of nitrosification bacteria (-70.94±26.17%) and nitrifiers (-47.86±23.68%) in biofilms which lead to significant reduction of ammoxidation in water (P<0.05). The N-cycling intensity from PC systems was affected more strongly by E. coli than were MA systems. Furthermore, the coupling coefficient of exogenous E. coli to indigenous N-cycling bacteria in sediments (6.061, average connectivity degree) was significantly weaker than that in biofilms (9.852). Additionally, at the genus level, E. coli were most-closely associated with N-cycling bacteria such as Prosthecobacter, Hydrogenophaga, and Bacillus in sediments and biofilms according to co-occurrence bacterial network (Spearman). E. coli directly changed their abundance, so that the variability of species composition of N-cycling bacterial taxa was triggered, as well. Overall, exogenous E. coli repressed ammoxidation, but promoted ammonification and denitrification. Our results provided new insights into how pathogens influence the nitrogen cycle in freshwater ecosystems.

Structure and Functions of Hydrocarbon-Degrading Microbial Communities in Bioelectrochemical Systems

Bioelectrochemical systems (BESs) exploit the interaction between microbes and electrodes. A field of application thereof is bioelectrochemical remediation, an effective strategy in environments where the absence of suitable electron acceptors limits classic bioremediation approaches. Understanding the microbial community structure and genetic potential of anode biofilms is of great interest to interpret the mechanisms occurring in BESs. In this study, by using a whole metagenome sequencing approach, taxonomic and functional diversity patterns in the inoculum and on the anodes of three continuous-flow BES for the removal of phenol, toluene, and BTEX were obtained. The genus Geobacter was highly enriched on the anodes and two reconstructed genomes were taxonomically related to the Geobacteraceae family. To functionally characterize the microbial community, the genes coding for the anaerobic degradation of toluene, ethylbenzene, and phenol were selected as genetic markers for the anaerobic degradation of the pollutants. The genes related with direct extracellular electron transfer (EET) were also analyzed. The inoculum carried the genetic baggage for the degradation of aromatics but lacked the capacity of EET while anodic bacterial communities were able to pursue both processes. The metagenomic approach provided useful insights into the ecology and complex functions within hydrocarbon-degrading electrogenic biofilms.

Dynamic responses of community structure and microbial functions of periphytic biofilms during chronic exposure to TiO2 NPs

We investigated the dynamic response of community structure and microbial function of periphytic biofilms during long-term exposure to TiO2 NPs.

Membrane autopsy deciphering keystone microorganisms stubborn against online NaOCl cleaning in a full-scale MBR

The knowledge about membrane biofouling evolution in full-scale membrane bioreactor (MBR) applications is quite lacking, notwithstanding a few lab-scale investigations. For the first time, this study elaborated the effect of online NaOCl cleaning on the dynamic development of membrane biofilm microbiota during long-term operation of a large-scale MBR for municipal wastewater treatment (40,000 m3/d). Four times of membrane autopsies were conducted during 160 days operation to scrutinize the microbial community and concomitant organic foulants. The transmembrane pressure difference (TMP) development revealed limited effect of 30 min online NaOCl cleaning on long-term biofouling removal. NaOCl not only altered the structure of biofilm communities but also increased the richness and evenness on early fouling stages. Meanwhile, network analysis revealed the keystone taxa f_Comamonadaceae that played key roles in stabilizing community structure and developing anti-cleaning and irreversible fouling propensity of the biofilm. NaOCl cleaning also impacted the evolving of keystone taxa by intensifying the competition between the dominated taxa f_Moraxellaceae and other species during early fouling stages. Furthermore, the succession of the biofilm microbiota synchronously accelerated the TMP increase and the accumulation of organic foulants including polysaccharides, aromatic proteins and soluble microbial products during biofilm maturation. These identified key stubborn foulants shed light on limitations of current online NaOCl cleaning and provide guidance to optimize the efficiency of online chemical cleaning protocols in full-scale MBR operations.

Seasonality combined with the orientation of surfaces influences the microbial community structure of biofilms in the deep Mediterranean Sea

Seasonality combined with the orientation of surfaces influences the microbial community structure of biofilms in the deep Mediterranean Sea

Seasonality combined with the orientation of surfaces influences the microbial community structure of biofilms in the deep Mediterranean Sea

Attachment to surfaces represents an important life strategy for microbial communities as indicated by the rapid colonization of biotic and abiotic surfaces in marine waters. However, little attention has been paid to the development of biofilm-associated microbial communities and the environmental parameters influencing biofilm development in the deep sea. In this study, a deep-sea experimental setup was used to follow the development of the microbial community colonizing solid surfaces deployed at 4500 m depth at the deepest point of the Mediterranean Sea. The experiment was performed during summer (May to October 2007) and winter (October 2007–May 2008), each lasting for 155 d. The phylogenetic composition of the biofilm community was determined by tag sequencing of the 16S rRNA gene. We investigated whether the composition of the deep-sea microbial biofilms is influenced by seasonality. Based on tag sequencing, operational taxonomic units were identified and diversity indices calculated. Seasonality combined with the orientation of the solid surface on which the biofilms were growing was the main factor influencing the structure of the microbial community. The most abundant phyla of deep-sea biofilm communities attached to the solid surfaces were Gammaproteobacteria (range: 10.8%–92.6%), Alphaproteobacteria (range: 34.9%–92.6%) and Betaproteobacteria (range: 0.3%–2.1%), irrespective of the variables (surface, orientation, season). Flavobacteria and Epsilonbacteria show a clear preference with respect to the orientation of the deployed surfaces during the winter, however, they were essentially absent at the surfaces during the summer. Some bacterial classes such as Campylobacterales and Rhodobacterales showed distinct preferences for specific seasons or orientation of the substrate. Taken together, we conclude that even on deep-sea biofilms, there is to some extent seasonality detectable in the composition of the surface associated prokaryotic community, despite the fact that the deep-sea is, in terms of physico-chemical parameters, a fairly stable environment.

Burial depth of anode affected the bacterial community structure of sediment microbial fuel cells

Sediment microbial fuel cells (SMFCs) are attractive devices to in situ power environmental monitoring sensors and bioremediate contaminated soils/sediments. Burial depth of the anode was verified to affect the performance of SMFCs. The present research evaluated the differences in microbial community structure of anodic biofilms located at different depth. It was demonstrated that both microbial diversity and community structure of anodic biofilms were influenced by the depth of anode location. Microbial diversity decreased with increased anodic depth. The number of the operational taxonomic units (OTUs) was determined as 1438 at the anode depth of 5 cm, which reduced to 1275 and 1005 at 10 cm and 15 cm, respectively. Cluster analysis revealed that microbial communities of 5 cm and 10 cm were clustered together, separated from the original sediment and 15 cm. Proteobacteria was the predominant phylum in all samples, followed by Bacteroidetes and Firmicutes. Beta-and Gamma-proteobacteria were the most abundant classes. A total of 23 OTUs showed high identity to 16S rRNA gene of exoelectrogens such as Geobacter and Pseudomonas. The present results provided insights into the effects of anode depth on the performance of SMFC from the perspectives of microbial community structure.

Biofilm pads—an easy method to manufacture artificial biofilms embedded in an alginate polymer matrix

AbstractPeriphytic biofilms are the major resource for many herbivorous invertebrates in both marine and freshwater benthos. They are of crucial importance for benthic food webs, substrate stability, and biogeochemical processes in littoral zones. While the importance of invertebrate grazing on biofilms has been studied extensively using natural, mixed algal communities grown on artificial substrates, there is so far no method available to create defined periphyton communities for these grazing studies. The reason for this is that many benthic algae interact with co‐occurring species within the extracellular polymeric substances (EPSs) that form the nonorganismic part of the biofilm. Here, we present a novel method that allows the manufacturing of defined monoculture and multispecies biofilms by using an alginate polymer as artificial EPS structure, into which algal cultures can be embedded. Using confocal laser scanning microscopy, we show that alginate effectively embeds various algal taxa in an EPS matrix that is very similar to natural biofilms. In a grazing experiment, we demonstrate that several common freshwater herbivorous invertebrates can graze these alginate biofilms efficiently. As the method is easy to handle, it allows for highly controlled feeding experiments with benthic herbivores to assess, for example, the role of algal biodiversity on the efficiency of top‐down control, the effects of environmental drivers such as nutrients, salinity, or seawater acidification on biofilm community structure, and the impacts of herbivory in benthic communities.

Intimately coupled TiO2/g-C3N4 photocatalysts and in-situ cultivated biofilms enhanced nitrate reduction in water

Nitrogen pollution has become a major threat to the aquatic environment, and the key to solve this problem is to enhance the nitrate reduction in water. Intimately coupled photocatalysis and biodegradation (ICPB) is a novel technique for degradation of bio-recalcitrant pollutants; however, it has not been used for nitrate reduction. This study applied a system that intimately coupled TiO2/g-C3N4 photocatalysts and in-situ cultivated biofilms for nitrate reduction in water without additional electron donors. The result shows a nitrate removal rate up to 40.3% in ICPB after 16 h. Furthermore, the N2 selectivity was 86.3%, which was higher than that for other protocols. In ICPB, nitrate was first reduced to nitrite, which was mainly contributed to the effect of biofilms. Then, nitrite was reduced via the cooperation of photocatalysts and biofilms, which benefited both low production of undesirable products and high selectivity of N2. Moreover, the growth of the microbes during the ICPB were favorable for denitrification. In conclusion, light-induced electrons generated from TiO2/g-C3N4 were harvested by microbes, optimizing the community structure of biofilms toward nitrate reduction. This study provides a new strategy to enhance the nitrate reduction in water and an insight into the reduction mechanism of nitrate in ICPB.

Inhibition of microbial influenced corrosion on carbon steel ST37 using biosurfactant produced by Bacillus sp.

Microbial influenced corrosion that associated with biofilm has been one of the major problem in industry. It causes damage to equipment and infrastructure because of rapid material deterioration and often resulting in pipeline failures. Pipeline failures can lead to large economic losses and environmental problems. Synthetic chemical biocides are commonly used to prevent corrosion but are not effective against preformed biofilm on pipes and toxic to the environment. A new antimicrobial is being developed by using biosurfactant produced by indigenous oil reservoir bacteria Bacillus sp. to prevent and eradicate biofilm. This study aims to determine minimum inhibitory concentration (MIC), minimum biofilm inhibitory concentration (MBIC), and minimum biofilm eradication concentration for 50% eradication (MBEC50) of biosurfactant against biofilm forming bacteria isolated from oil resevoir, its effect on biofilm community structure and its ability to inhibit the corrosion rate of Carbon Steel ST37. MIC, MBIC, and MBEC50 were determined using broth macrodillution technique, biofilm community structure were analyzed using total plate count method, and corrosion rate of steel were determined using weight loss method. Biofilm and corroded steel surface was also visualized using SEM-EDS. From this study, it is revealed that MIC, MBIC, and MBEC50 values respectively are 62.5; 31.25; and 500 μg ml−1. Biosufactant is able to inhibit Pseudomonas sp. 1 and Pseudomonas sp. 2 attachment to Carbon Steel ST37 surface and also able to eradicate preformed biofilm on steel surface. This study also showed the reduction of corrosion rate in Carbon Steel ST 37 as a result of biosurfactants treatment; from 4.56 × 10−4 mm year−1 to 3.31 × 10−4 mm year−1 based on MBIC value and from 5.18 × 10−5 mm year −1 to 2.7 × 10−5 mm year−1 after biofilm eradication at MBEC value. The results showed that biosurfactant in this study could be a good candidate for a new anti-corrosion agent.

Ecology of Subseafloor Crustal Biofilms.

The crustal subseafloor is the least explored and largest biome on Earth. Interrogating crustal life is difficult due to habitat inaccessibility, low-biomass and contamination challenges. Subseafloor observatories have facilitated the study of planktonic life in crustal aquifers, however, studies of life in crust-attached biofilms are rare. Here, we investigate biofilms grown on various minerals at different temperatures over 1–6 years at subseafloor observatories in the Eastern Pacific. To mitigate potential sequence contamination, we developed a new bioinformatics tool – TaxonSluice. We explore ecological factors driving community structure and potential function of biofilms by comparing our sequence data to previous amplicon and metagenomic surveys of this habitat. We reveal that biofilm community structure is driven by temperature rather than minerology, and that rare planktonic lineages colonize the crustal biofilms. Based on 16S rRNA gene overlap, we partition metagenome assembled genomes into planktonic and biofilm fractions and suggest that there are functional differences between these community types, emphasizing the need to separately examine each to accurately describe subseafloor microbe-rock-fluid processes. Lastly, we report that some rare lineages present in our warm and anoxic study site are also found in cold and oxic crustal fluids in the Mid-Atlantic Ridge, suggesting global crustal biogeography patterns.

Shear Stress as a Major Driver of Marine Biofilm Communities in the NW Mediterranean Sea.

While marine biofilms depend on environmental conditions and substrate, little is known about the influence of hydrodynamic forces. We tested different immersion modes (dynamic, cyclic and static) in Toulon Bay (north-western Mediterranean Sea; NWMS). The static mode was also compared between Toulon and Banyuls Bays. In addition, different artificial surfaces designed to hamper cell attachment (self-polishing coating: SPC; and fouling-release coating: FRC) were compared to inert plastic. Prokaryotic community composition was affected by immersion mode, surface characteristics and site. Rhodobacteriaceae and Flavobacteriaceae dominated the biofilm community structure, with distinct genera according to surface type or immersion mode. Cell density increased with time, greatly limited by hydrodynamic forces, and supposed to delay biofilm maturation. After 1 year, a significant impact of shear stress on the taxonomic structure of the prokaryotic community developed on each surface type was observed. When surfaces contained no biocides, roughness and wettability shaped prokaryotic community structure, which was not enhanced by shear stress. Conversely, the biocidal effect of SPC surfaces, already major in static immersion mode, was amplified by the 15 knots speed. The biofilm community on SPC was 60% dissimilar to the biofilm on the other surfaces and was distinctly colonized by Sphingomonadaceae ((Alter)Erythrobacter). At Banyuls, prokaryotic community structures were more similar between the four surfaces tested than at Toulon, due possibly to a masking effect of environmental constraints, especially hydrodynamic, which was greater than in Toulon. Finally, predicted functions such as cell adhesion confirmed some of the hypotheses drawn regarding biofilm formation over the artificial surfaces tested here.

Deterioration and microbial colonization of cultural heritage stone buildings in polluted and unpolluted tropical and subtropical climates: A meta-analysis

Climatic factors, mainly rainfall, temperature and relative humidity, air pollution and biological activity influence the processes of stone degradation. This article analyses results of studies on microbial (mainly cyanobacterial) colonization of 32 cultural heritage stone monuments in Argentina, Belize, Brazil, Cambodia, Laos and Mexico, performed by four of the six authors over several years. Principal components analysis (PCA) and canonical analysis of principal coordinates (CAP) revealed that stone type, calcareous or siliceous, affected cyanobacterial community structure in biofilms. Köppen climate class was important, more temperate climates grouping separately from more tropical sites. Highly polluted cities showed reduced cyanobacterial diversity, with centroid completely separate from the rest; Chroococcus appeared to be relatively pollution-resistant. Buildings at marine sites showed a different and somewhat reduced cyanobacterial diversity. The genus Gloeocapsa was the most frequent genus detected overall, present at all but two sites. Various cases are discussed in more detail, along with the accompanying biodeterioration, and special mention is made of black crusts seen on some buildings. Although some next generation sequencing analyses are discussed, the statistical analyses are based on results of standard microbiological techniques that have been in use for many years and were employed similarly in all the studies.

Unraveling interactive characteristics of microbial community associated with bioelectric energy production in sludge fermentation fluid-fed microbial fuel cells

This first-attempt study deciphered the interactive characteristics of anodophilic microbial community-associated bioelectricity production in waste activated sludge (WAS) fermentation fluid-fed microbial fuel cells (MFCs). A novel schematic elucidation for illustrating synergistic interactions in anodic microbial consortia towards electrogenesis was proposed. Moreover, the specific genera of Pseudomonas, Desulfovibrio, Phyllobacterium, Desulfuromonas, Chelatococcus and Aminivibrio were dominant in anodic biofilms, leading to an electrogenesis efficiency of 1.254 kWh/kg COD and peak power density of 0.182 W/m2 (at feeding level of 1.20 g COD/L). It was apparently higher than those MFCs fed with glucose/acetate. The fermentative species contributed positively in reorganizing microbial community structure in anodic biofilms, positively relating to electrogenesis via interactions with exoeletrogens in MFCs. Finally, a more electrogenesis was positively associated to larger anodic microbial diversity, relatively medium anodic community evenness, together with higher abundance of functional genes related to electrogenesis in functional species in MFCs fed with WAS fermentation fluid.

Microbial biofilm formation and community structure on low-density polyethylene microparticles in lake water microcosms

The occurrence of microplastics (MPs) in the environment has been gaining widespread attention globally. MP-colonizing microorganisms are important links for MPs contamination in various ecosystems, but have not been well understood. To partially address this issue, the present study investigated biofilm formation by microorganisms originating from lake water on low-density polyethylene (LDPE) MPs using a cultivation approach and the surface-related effects on the MP-associated microbial communities using 16S rRNA high-throughput sequencing. With the addition of nonionic surfactants and UV-irradiation pretreatment that changed the surface properties of LDPE MPs, more microorganisms were colonized on LDPE surface. Microbial community analysis indicated that LDPE MPs were primarily colonized by the phyla Proteobacteria, Bacteroidetes and Firmicutes, and the surface roughness and hydrophobicity of MP were important factors shaping the LDPE MP-associated microbial community structure. Half of the top 20 most abundant genera colonizing on LDPE were found to be potential pathogens, e.g., plant pathogens Agrobacterium, nosocomial pathogens Chryseobacterium and fish pathogens Flavobacterium. This study demonstrated rapid bacterial colonization of LDPE MPs in lake water microcosms, the role of MPs as transfer vectors for harmful microorganisms in lake water, and provided a first glimpse into the effect of surface properties on LDPE MP-associated biofilm communities.

Impact Mechanisms of Carboxyl Group Modified Cathode on Acetate Production in Microbial Electrosynthesis Systems

Microbial electrosynthesis systems (MESs) can convert carbon dioxide into added value compounds using microorganisms as catalyst, which is expected to help achieve conversion of greenhouse gases into resources. However, the synthetic efficiency of MESs is far behind the industry requirements. In this study, carbon cloth surfaces were bonded with carboxyl groups by electrochemical reduction of aryl diazonium salts and then used as a cathode in MESs reactors. The results showed that the hydrophilicity of the carbon cloth surfaces improved after the carboxyl groups were modified. However, weaker current of cyclic voltammetry was obtained in the modified cathode. Significant differences were observed between modified (CA-H, CA-M, CA-L) and non-modified cathode (CK) during the start-up period. After 48h, the hydrogen production rate of CA-H, CA-M, CA-L was 21.45, 28.60, and 22.75 times higher than CK. After 120h, the acetate accumulation concentration of CA-H, CA-M, CA-L was 2.01, 2.43, and 1.44 times higher than CK. After 324h, there was little difference in the electrochemical activity of cathodic biofilm and protein content (about 0.47 mg·cm-2) in all groups. The analysis of the community structure of cathodic biofilm showed that, in the genus level, Acetobacterium, Norank_p_Saccharibacteria, and Thioclava were the dominant species, accounting for 59.6% to 82.1%. There was little difference in the relative abundance of Acetobacterium in all groups (31.3% to 40.1%). However, the relative abundance of norank_p_Saccharibacteria in CA-H, CA-M, CA-L, and CK were 16.1%, 24.6%, 31.1%, and 37.5%, respectively. The carboxyl modified cathode had a great influence on the start-up stage of MESs, which could be a new idea for the rapid start-up of MESs.

Dynamics of the physiochemical and community structures of biofilms under the influence of algal organic matter and humic substances

Increased loading of algal organic matter (AOM) during harmful algal blooms not only burdens water treatment processes but also challenges safe drinking water delivery. While organic constituents promote biofilm growth in drinking water distribution systems (DWDS), the effects of AOM on biofilm formation in DWDS are not well understood. Herein, three parallel biofilm reactors were used to assess and compare how treated AOM- and humic substance (HS)-impacted bulk water, and R2A medium (a control) affect biofilm development for 168 days. The 16S rRNA gene sequencing analysis revealed that the bacterial communities in biofilms were clustered with the organic matter types in bulk water, where Family Comamonadaceae was the most dominant but showed different temporal dynamics depending on the organic matter characteristics in bulk water. Higher diversity was observed in the biofilms grown in AOM-impacted bulk water (BFAOM) than biofilms grown in HS-impacted (BFHS) and R2A-impacted bulk water (BFR2A) as the biofilms matured. In addition, some taxa (e.g., Rhodobacteraceae and Sphingomonadaceae) were enriched in BFAOM compared to BFHS and BFR2A. The biofilm image analysis results indicated that compared to BFHS, BFAOM and BFR2A had relatively thinner and heterogeneous physical structures with lower amounts of cell biomass, extracellular polymeric substances (EPS), and higher EPS protein/polysaccharide ratios. Overall, this study revealed how AOM- and HS-impacted bulk water shape the physiochemical and community structures of biofilms, which can provide insights into assessing biofilm-associated risks and optimizing disinfection practices for biofilm control in DWDS.