Biofilm Community Structure Research Articles

Redox potential shapes spatial heterogeneity of mixed-cultured electroactive biofilm treating wastewater

The core of bioelectrochemical systems (BESs) is electrochemically active microorganisms (EAMs), which exert spatial heterogeneity on electrode surface and influences BESs performance. Setting an optimal potential is an effective strategy for improving and optimizing BESs performance, however, how the electrode potential affects spatial structure of microbial community within anode biofilm is not known. Using a complex substrate-fed BES with a wastewater inoculum, this study investigated the community structure and composition of the stratified biofilm developed under the potential of −0.3 V, 0 V, +0.3 V and +0.6 V (vs. saturated calomel electrode) by freezing microtome method and high-throughput sequencing analysis. The spatial heterogeneity of biofilm community was found to be dependent on the electrode potential and a less stratified community structure was observed for +0.6 V than other potentials. Within the biofilms, the inner layers selected more Geobacter and the outer layers enriched more Acinetobacter and Serratia, potentially suggested a stratification of electron transfer pathway and metabolite-based interspecies communications. The results demonstrated the response of spatial heterogeneity of anode biofilm community to the change of electrode potential, which helps to understand the selectivity and enrichment of kinetically efficient anodic microbiome by electron potential.

Effects of gradual increase of ciprofloxacin and cefotaxime on nitrogen and phosphorus removal and microbial community in moving bed biofilm reactor

To clarify the effects of trace amount of antibiotics in wastewater on biofilm reactor, this study explored the effects of gradual increase of ciprofloxacin (CIP) and cefotaxime (CTX) ranging from 100 to 1000 ng·L−1 on the nitrogen and phosphorus removal and microbial community in a moving bed biofilm reactor (MBBR). Under different phases of CIP and CTX gradual increase, the resilience of culturable functional bacteria including ammonifying bacteria, nitrite oxidizing bacteria, aerobic denitrifying bacteria and phosphorus accumulating organisms in the biofilm was analyzed. High-throughput sequencing analysis was used to reveal the microbial community structure and species composition of functional bacteria. The results indicated that the treatment performance of MBBR was temporarily affected by trace CIP and CTX and returned to its original level within 1–2 days. Ammonifying bacteria exhibited strong resilience in trace CIP and CTX stress environments. Trace CIP and CTX altered the microbial community structure in biofilm, but the performance of nitrogen and phosphorus removal in MBBR was slightly affected. This might attribute to the rapid proliferation of Thauera, Dechloromonas and Candidatus_Accumulibacter on the carrier biofilm. The research results help to deepen the understanding of the impact of trace antibiotics on the MBBR and to provide a reference for the development of microbial germplasm resources for biological nitrogen and phosphorus removal from wastewater under antibiotic stress conditions.

Bacterial diversity in the biofilms on mariculture polystyrene foam at Xiamen’s coast

Expanded polystyrene (EPS), commonly used as floating material in mariculture, could be a significant source of coastal PS waste. Prolonged floating in seawater leads to the formation of dense biofilms, which could disperse into the oceanic environment and spread to water bodies even the seabed as EPS deteriorates into microplastics. To understand the bacterial community structure in EPS surface biofilms, this study employed high-throughput sequencing technology to analyze the biofilms on EPS foams used in offshore mariculture aside Xiamen. Analysis of 24 biofilm samples from different seasons revealed significant differences in bacterial community structures. However, we have identified the core groups of the EPS biofilm, including Flavobacteriaceae, Rhodobacteraceae, Halomonadaceae, Exiguobacteriaceae, Vibrionaceae, and Pseudoalteromonadaceae. The Halomonadaceae and Exiguobacteriaceae are particularly attracted to the subtropical EPS material. In addition, groups with potential of degrading PS were identified, such as Alcanivorax, Pseudomonas, Acinetobacter, Exiguobacterium, Brevundimonas, and Bacillus, as well as the potential fish pathogen Acinetobacter johnsonii. The microbial biofilms formed on floating EPS demonstrate high diversity and are strongly influenced by environmental conditions. This study contributes to a deeper understanding of the interactions of bacteria in the biofilm with widely used and dispersed floating materials in the oceans.

Effect of immersion time on bacterial community structure and sulfur metabolism in biofilm on concrete surface in sewer environment

This study focuses on an overlooked but critical issue: the composition and functional expression of microbial communities on the concrete surface in different areas of sewer pipes. Three immersion conditions were applied to simulate the duration of concrete in different areas ofsewers exposed to sewage, including short-term (L1), long-term (L2) and permanent immersion (L3). The properties of concrete under different immersion conditions and the bacterial diversity and functional capabilities in biofilms on the concrete surface were analyzed. Results showed that the L1 group was dominated by Halothiobacillus, whereas Desulfomicrobium was prominent in the L3 group. Significant differences in the predominant functional microbial communities and metabolic functional genes further confirmed the strong impact of immersion time on the pathways of microbial sulfur metabolism and concrete performance in sewer environment. Compared with the L2 and L3 groups, the decreased sewage immersion time resulted in an increase in the abundance and metabolic activity of sulfur-oxidizing bacteria in the L1 group. Hence, greater mass loss and gypsum production of concrete was found in the L1 group. The structural and functional differentiation of bacterial communities on the concrete surface observed in this study contributes to a better understanding of the uneven corrosion in real sewer pipes.

Mechanisms of partial denitrification in an integrated fixed-film activated sludge (IFAS) system: From progressive startup to stabilization

The rapid startup and long-term stability of partial denitrification (PD) processes are essential prerequisites for successful PD/Anammox systems. Bioaggregates play a significant role in impacting mass transfer efficiency and shaping the microbial community structure within the PD process. Thus, the startup and stable operation of the PD process were investigated in an integrated fixed-film activated sludge (IFAS) by inoculating activated sludge and biofilm from anoxic tanks of a wastewater treatment plant. During the initial stage of startup, a sudden increase in NO3−-N concentration induced a rapid enzyme stress response, leading to a significant increase in total NaR activity (1.38 ± 0.0 μmol·(L·min)−1 to 25.94 ± 0.23 μmol·(L·min)−1), concomitant with the inhibition of total NiR activity (13.93 ± 0.48 μmol·(L·min)−1 to 4.25 ± 0.26 μmol·(L·min)−1), thereby inducing the rapid accumulation of NO2−-N (13.39 mg·L−1) within 6 days. During the later stage of startup, enzyme stress response was subsided but the partial denitrifiers were gradually enriched, consequently leading to continuous accumulation of NO2−-N. The results of ex-situ activity showed that flocs exhibit superior PD performance in contrast to biofilm throughout the entire operational period. Meanwhile, the total extracellular polymeric substances (EPS) content and community structure in both flocs and biofilm indicated that the migration of EPS-secreted partial denitrifiers (Thauera) from flocs to biofilm was obliged by substrate deficiency during the stable operating stage (days 35–100). Therefore, the startup of PD may be sequentially mediated by two distinct mechanisms, enzyme stress response and functional bacterial enrichment, with microbial community migration occurring during the stable phase.

Biofilm of petroleum-based and bio-based microplastics in seawater in response to Zn(II): Biofilm formation, community structure, and microbial function

Microplastic biofilms are novel vectors for the transport and spread of pathogenic and drug-resistant bacteria. With the increasing use of bio-based plastics, there is an urgent need to investigate the microbial colonization characteristics of these materials in seawater, particularly in comparison with conventional petroleum-based plastics. Furthermore, the effect of co-occurring contaminants, such as heavy metals, on the formation of microplastic biofilms and bacterial communities remains unclear. In this study, we compared the biofilm bacterial community structure of petroleum-based polyethylene (PE) and bio-based polylactic acid (PLA) in seawater under the influence of zinc ions (Zn2+). Our findings indicate that the biofilm on PLA microplastics in the late stage was impeded by the formation of a mildly acidic microenvironment resulting from the hydrolysis of the ester group on PLA. The PE surface had higher bacterial abundance and diversity, with a more intricate symbiotic pattern. The bacterial structures on the two types of microplastics were different; PE was more conducive to the colonization of anaerobic bacteria, whereas PLA was more favorable for the colonization of aerobic and acid-tolerant species. Furthermore, Zn increased the proportion of the dominant genera that could utilize microplastics as a carbon source, such as Alcanivorax and Nitratireductor. PLA had a greater propensity to harbor and disseminate pathogenic and drug-resistant bacteria, and Zn promoted the enrichment and spread of harmful bacteria such as, Pseudomonas and Clostridioides. Therefore, further research is essential to fully understand the potential environmental effects of bio-based microplastics and the role of heavy metals in the dynamics of bacterial colonization.

Interspecies interactions in dairy biofilms drive community structure and response against cleaning and disinfection

Interspecies interactions within a biofilm community influence population dynamics and community structure, which in turn may affect the bacterial stress response to antimicrobials. This study was conducted to assess the impact of interactions between Kocuria salsicia and a three-species biofilm community (comprising Stenotrophomonas rhizophila, Bacillus licheniformis, and Microbacterium lacticum) on biofilm mass, the abundance of individual species, and their survival under a laboratory-scale cleaning and disinfection (C&D) regime. The presence of K. salsicia enhanced the cell numbers of all three species in pairwise interactions. The outcomes derived from summing up pairwise interactions did not accurately predict the bacterial population dynamics within communities of more than two species. In four-species biofilms, we observed the dominance of S. rhizophila and B. licheniformis, alongside a concurrent reduction in the cell counts of K. salsicia and M. lacticum. This pattern suggests that the underlying interactions are not purely non-transitive; instead, a more complex interplay results in the dominance of specific species. We observed that bacterial spatial organization and matrix production in different mixed-species combinations affected survival in response to C&D. Confocal microscopy analysis of spatial organization showed that S. rhizophila localized on the biofilm formed by B. licheniformis and M. lacticum, and S. rhizophila was more susceptible to C&D. Matrix production in B. licheniformis, evidenced by alterations in biofilm mass and by scanning electron microscopy, demonstrated its protective role against C&D, not only for this species itself, but also for neighbouring species. Our findings emphasise that various social interactions within a biofilm community not only affect bacterial population dynamics but also influence the biofilm community's response to C&D stress.

Oxygen-dependent biofilm dynamics in leaf decay: an in vitro analysis

Biofilms are important in the natural process of plant tissue degradation. However, fundamental knowledge of biofilm community structure and succession on decaying leaves under different oxygen conditions is limited. Here, we used 16S rRNA and ITS gene amplicon sequencing to investigate the composition, temporal dynamics, and community assembly processes of bacterial and fungal biofilms on decaying leaves in vitro. Leaves harvested from three plant species were immersed in lake water under aerobic and anaerobic conditions in vitro for three weeks. Biofilm-covered leaf samples were collected weekly and investigated by scanning electron microscopy. The results showed that community composition differed significantly between biofilm samples under aerobic and anaerobic conditions, though not among plant species. Over three weeks, a clear compositional shift of the bacterial and fungal biofilm communities was observed. The alpha diversity of prokaryotes increased over time in aerobic assays and decreased under anaerobic conditions. Oxygen availability and incubation time were found to be primary factors influencing the microbial diversity of biofilms on different decaying plant species in vitro. Null models suggest that stochastic processes governed the assembly of biofilm communities of decaying leaves in vitro in the early stages of biofilm formation and were further shaped by niche-associated factors.

Biotic and abiotic factors interplay in structuring the dynamics of microbial co-occurrence patterns in tropical mountainsides

Ecological interactions are important for maintaining biodiversity and ecosystem functions. Particularly in stream biofilms, little is known about the distributional patterns of different taxonomic groups and their potential interactions along elevational gradients. Here, we investigated the bacterial and fungal community structures of stream biofilms across elevational gradients on Mount Kilimanjaro, and explored patterns of their distribution, diversity, community structures, and taxa co-occurrence. We found that fungal and bacterial richness were more convergent at higher elevations, while their community structures became significantly more divergent. Inferred network complexity and stability significantly decreased with increasing elevation for fungi, while an opposite trend was observed for bacteria. Further quantitative analyses showed that network structures of bacteria and fungi were more divergent as elevation increased. This pattern was strongly associated with shifts in abiotic factors, such as mean annual temperatures, water PO43--P, and stream width. By constructing bipartite networks, we showed the fungal-bacterial network to be less redundant, more clustering, and unstable with increasing elevation. Abiotic factors (e.g., temperatures and stream width) and microbial community properties (i.e., structure and composition) significantly explained the dynamic changes in fungal-bacterial network properties. Taken together, this study provides evidence for the interplay of biotic and abiotic factors structuring potential microbial interactions in stream biofilms along a mountainside elevational gradient.

The role of phages for microdiverse bacterial communities in proglacial stream biofilms

Viruses modulate the diversity and activity of microbial communities. However, little is known about their role for the structure of stream bacterial biofilm communities. Here, we present insights into the diversity and composition of viral communities in various streams draining three proglacial floodplains in Switzerland. Proglacial streams are characterized by extreme environmental conditions, including near-freezing temperatures and ultra-oligotrophy. These conditions select for few but well-adapted bacterial clades, which dominate biofilm communities and occupy niches via microdiversification. We used metagenomic sequencing to reveal a diverse biofilm viral assemblage in these streams. Across the different floodplains and streams, viral community composition was tightly coupled to that of the bacterial hosts, which was underscored by generally high host specificity. Combining predictions of phage-host interactions with auxiliary metabolic genes (AMGs), we identify specific AMGs shared by phages infecting microdiverse clade members. Our work provides a step towards a better understanding of the complex interactions among bacteria and phages in stream biofilm communities in general and streams influenced by glacier meltwaters and characterized by microdiversity in particular.

Microbial community structure and co-occurrence network stability in seawater and microplastic biofilms under prometryn pollution in marine ecosystems

Prometryn has been extensively detected in marine environment because of its widespread usage in agriculture and aquaculture and has been concerns since its serious effects on aquatic organisms. However, its impact on the microbial community in the marine ecosystem including seawater and biofilm is still unclear. Therefore, a short-term indoor microcosm experiment of prometryn exposure was conducted. This study found that prometryn had a more significant impact on the structure and stability of the microbial community in seawater compared to microplastic biofilms. Additionally, we observed that the assembly of the microbial community in biofilms was more affected by stochastic processes than in seawater under the exposure of prometryn. Our study provided evidence for the increasing impact of the microbial communities under the stress of prometryn and microplastics.

Microbial Diversity and Community Structure of Wastewater-Driven Microalgal Biofilms.

Dwindling water sources increase the need for efficient wastewater treatment. Solar-driven algal turf scrubber (ATS) system may remediate wastewater by supporting the development and growth of periphytic microbiomes that function and interact in a highly dynamic manner through symbiotic interactions. Using ITS and 16S rRNA gene amplicon sequencing, we profiled the microbial communities of four microbial biofilms from ATS systems operated with municipal wastewater (mWW), diluted cattle and pig manure (CattleM and PigM), and biogas plant effluent supernatant (BGE) in comparison to the initial inocula and the respective wastewater substrates. The wastewater-driven biofilms differed significantly in their biodiversity and structure, exhibiting an inocula-independent but substrate-dependent establishment of the microbial communities. The prokaryotic communities were comparable among themselves and with other microbiomes of aquatic environments and were dominated by metabolically flexible prokaryotes such as nitrifiers, polyphosphate-accumulating and algicide-producing microorganisms, and anoxygenic photoautotrophs. Striking differences occurred in eukaryotic communities: While the mWW biofilm was characterized by high biodiversity and many filamentous (benthic) microalgae, the agricultural wastewater-fed biofilms consisted of less diverse communities with few benthic taxa mainly inhabited by unicellular chlorophytes and saprophytes/parasites. This study advances our understanding of the microbiome structure and function within the ATS-based wastewater treatment process.

Climate Change and Plastic Pollution: A Review of Their Connections

The world faces two major environmental issues concurrently, namely climate change and plastic pollution. Though seemingly unrelated, they intricately influence each other. This review aims to present the intricate connections between climate change and plastic pollution through the review of recent literature in these genres. The review explains that global warming could increase plastic degradation through physical, chemical, and biological processes, leading to an increased abundance of microplastics. Global warming enhances the leaching of chemicals from microplastics. Higher temperatures promote desorption of chemicals sorbed on plastics by providing the adsorbates with more kinetic energy to overcome attractions with the adsorbents. Higher temperatures can also promote biofilm formation and alter the microbial community structures of biofilms. Melting sea ice and glaciers associated with warming temperatures release the microplastics trapped in the environment. Sea-level rise and extreme weather events enhance the transfer of microplastics between land, ocean, and air, thus changing their distribution and transport, while ocean acidification may influence the biofouling of microplastics and increase the vulnerability of some corals to the impacts of microplastics. Plastic pollution, however, exacerbates climate change due to the release of greenhouse gases throughout the lifecycle of plastics. Microplastics also adversely affect the growth of microalgae, hence the ocean carbon cycle. Airborne microplastics can alter the energy balance of the Earth through scattering and absorbing radiation. This review suggests a circular economic approach to minimize waste, maximize the reuse and recycling of plastics, and promote the use of plastic substitutes to address both issues.

Enhancing anammox bacteria enrichment in integrated fixed-film activated sludge partial nitritation/anammox process via floc retention control

A promising technology for partial nitritation/anammox (PN/A) processes to treat ammonium wastewater is integrated fixed-film activated sludge (IFAS). For practical applications, achieving efficient enrichment of anammox bacteria (AnAOB) remains a challenge. In this study, membranes were temporarily used to separate solid and liquid components to induce changes in the mixed liquor suspended solids of the flocs. With membrane separation, AnAOB proliferated rapidly with a seven-fold increase in the maximum specific growth rate (μ) (from 0.009 to 0.072 d−1) and a three-fold increase in the nitrogen removal rate (from 0.91 to 3.20 kg N/(m3·d)). Moreover, microbial community analysis showed significant changes in bacterial species richness and diversity with and without membrane separation. Overall, the regulation of flocs significantly influenced the microbial community structure of both flocs and biofilms leading to improved nitrogen removal efficiency in the IFAS-PN/A system.

Effects of polyethylene microplastics with different particle sizes and concentrations on the community structure and function of periphytic biofilms

The widespread use of microplastics (MPs) inevitably lead to their release into aquatic environment, where they are likely to encounter the periphytic biofilms, leading to changes in microbial community structure and function. However studies on the toxicity of MPs with different particle sizes and concentrations to biofilms are still lacking. Here, the effects of polyethylene (PE) MPs with three sizes (10, 40 and 120 µm) and two concentrations (2 and 20 mg/L) on biofilm communities were investigated in microcosms over a 28-day incubation period. High-throughput sequencing showed that the alpha and beta diversity of biofilm community were significantly altered by MPs, depending on their size and concentration. Typically, larger particle sizes and higher concentrations resulted in more significant changes in biofilm community structure, indicating a stronger shading effect of larger particles. Compared to the controls, 120 µm MPs treatments significantly reduced the Chao1 and Shannon index of biofilm community regardless of the concentration, while 40 µm MPs reduced the Shannon index only at high concentrations. Moreover, MPs resulted in the changes in the abundances of bacterial communities, with the abundance of Cyanobacteria increasing significantly while that of Proteobacteria and Bacteroidetes decreasing in all treatments. Interestingly, functional analysis by FAPROTAX suggested that a decrease in the abundance of chemoheterotrophy genes and an increase in photoautotrophic- and nitrification-associated genes were observed after exposure to MPs, leading to potential changes of the carbon and nitrogen metabolism of biofilms. These results highlighted that MPs exposure had negative impacts on microbial community structure and function, which in turn affects microbial-mediated processes.

Identification of rare microbial colonizers of plastic materials incubated in a coral reef environment.

Plastic waste accumulation in marine environments has complex, unintended impacts on ecology that cross levels of community organization. To measure succession in polyolefin-colonizing marine bacterial communities, an in situ time-series experiment was conducted in the oligotrophic coastal waters of the Bermuda Platform. Our goals were to identify polyolefin colonizing taxa and isolate bacterial cultures for future studies of the biochemistry of microbe-plastic interactions. HDPE, LDPE, PP, and glass coupons were incubated in surface seawater for 11 weeks and sampled at two-week intervals. 16S rDNA sequencing and ATR-FTIR/HIM were used to assess biofilm community structure and chemical changes in polymer surfaces. The dominant colonizing taxa were previously reported cosmopolitan colonizers of surfaces in marine environments, which were highly similar among the different plastic types. However, significant differences in rare community composition were observed between plastic types, potentially indicating specific interactions based on surface chemistry. Unexpectedly, a major transition in community composition occurred in all material treatments between days 42 and 56 (p < 0.01). Before the transition, Alteromonadaceae, Marinomonadaceae, Saccharospirillaceae, Vibrionaceae, Thalassospiraceae, and Flavobacteriaceae were the dominant colonizers. Following the transition, the relative abundance of these taxa declined, while Hyphomonadaceae, Rhodobacteraceae and Saprospiraceae increased. Over the course of the incubation, 8,641 colonizing taxa were observed, of which 25 were significantly enriched on specific polyolefins. Seven enriched taxa from families known to include hydrocarbon degraders (Hyphomonadaceae, Parvularculaceae and Rhodobacteraceae) and one n-alkane degrader (Ketobacter sp.). The ASVs that exhibited associations with specific polyolefins are targets of ongoing investigations aimed at retrieving plastic-degrading microbes in culture.

Sub-MIC antibiotics influence the microbiome, resistome and structure of riverine biofilm communities.

The effects of sub-minimum inhibitory concentrations (sub-MICs) of antibiotics on aquatic environments is not yet fully understood. Here, we explore these effects by employing a replicated microcosm system fed with river water where biofilm communities were continuously exposed over an eight-week period to sub-MIC exposure (1/10, 1/50, and 1/100 MIC) to a mix of common antibiotics (ciprofloxacin, streptomycin, and oxytetracycline). Biofilms were examined using a structure-function approach entailing microscopy and metagenomic techniques, revealing details on the microbiome, resistome, virulome, and functional prediction. A comparison of three commonly used microbiome and resistome databases was also performed. Differences in biofilm architecture were observed between sub-MIC antibiotic treatments, with an overall reduction of extracellular polymeric substances and autotroph (algal and cyanobacteria) and protozoan biomass, particularly at the 1/10 sub-MIC condition. While metagenomic analyses demonstrated that microbial diversity was lowest at the sub-MIC 1/10 antibiotic treatment, resistome diversity was highest at sub-MIC 1/50. This study also notes the importance of benchmarking analysis tools and careful selection of reference databases, given the disparity in detected antimicrobial resistance genes (ARGs) identity and abundance across methods. Ultimately, the most detected ARGs in sub-MICs exposed biofilms were those that conferred resistance to aminoglycosides, tetracyclines, β-lactams, sulfonamides, and trimethoprim. Co-occurrence of microbiome and resistome features consistently showed a relationship between Proteobacteria genera and aminoglycoside ARGs. Our results support the hypothesis that constant exposure to sub-MICs antibiotics facilitate the transmission and promote prevalence of antibiotic resistance in riverine biofilms communities, and additionally shift overall microbial community metabolic function.

Organic fertilizer facilitates the soil microplastic surface degradation and enriches the diversity of bacterial biofilm

The land-use of organic fertilizers is considered as an important sustainable method for resource utilization, which may have an impact on the microplastic behaviors in the soil. Here, a 240-d dark culture experiment was conducted to reveal the degradation and biofilm characteristics of degradable and refractory granule microplastics in soil and soil-fertilizer systems. The results indicated that microplastics generally exhibited a weak weight loss as well as a specific etiolation on the surface after the culture, especially polyvinyl-chloride and polyhydroxyalkanoates (PHA). Increase in carbon-oxygen functional groups and the changes of oxygen/carbon ratios were noticed, which implied that oxidation and degradation occurred on the surface of microplastics during the cultural process. The changes were more intense on the degradable PHA, and the fertilized-soil treatment than those of the refractory microplastics and the pure soil. Moreover, the addition of organic fertilizers enriched the community diversity of bacterial biofilm on multiple microplastic surfaces. In this regard, the animal fertilizers provided a stronger effect than the plant fertilizers. Overall, the soil, fertilizer and microplastic types affected the community structure and diversity of bacterial biofilm. The outcomes of this study would provide a theoretical basis for the utilization of organic matters for agricultural soil applications.

Complete mineralization of 2,4-dichlorophenoxyacetic acid in a reduction and oxidation Synergistic Platform (ROSP)

Emissions of 2,4-dichlorophenoxyactic acid (2,4-D), one of the most widely used herbicides, pose serious risks to human and ecosystem health. Although biodegradation and abiotic chemical methods are available for 2,4-D removal, continuous treatment systems for 2,4-D removal are rare. We evaluated a Reduction and Oxidation Synergistic Platform (ROSP) that sequentially links an H2-based membrane catalyst-film reactor (H2-MCfR) and an O2-based membrane biofilm reactor (O2-MBfR). In continuous operation of the ROSP, 50 μM (or 11 ppm) of 2,4-D was completely mineralized with a total HRT of 12 h, yielding an effluent concentration < 0.2 μM 2,4-D (or < 44 ppb). In the H2-MCfR, over 80% of the influent 2,4-D was reductively hydrodechlorinated to phenoxyacetic acid (POA). The produced POA was the primary substrate supporting the co-oxidation of 2,4-D in the O2-MBfR. The biofilm community structure and genes encoding enzymes for POA and 2,4-D co-oxidation were identified through shallow metagenomic sequencing of the biofilm DNA samples. The dominant bacterial genus, Variovorax, and various mono-/dioxygenase-encoding genes related to aromatic ring hydrolysis, hydroxylation, and ring cleavage were associated with the removal of 2,4-D. In summary, reductive-dehalogenation products from the H2-MCfR become the primary substrate to enable the co-oxidation of the halogenated organic pollutants in the O2-MBfR, which does not require the addition of other organic materials. Therefore, the ROSP eliminates the need for an added organic substrate, which greatly reduces the demand for O2 and minimizes CO2 emissions.

Let's Talk about Slime; or Why Biofouling Needs More Attention in Sensor Science.

Although there is a growing demand for new sensors for environmental monitoring, biofouling continues to plague current sensors and sensing networks. As soon as a sensor is placed in water, the formation of a biofilm begins. Once a biofilm is established, reliable measurements are often no longer possible. Although current biofouling mitigation strategies can slow the biofouling process, a biofilm will eventually develop on or near the sensing surface. While antibiofouling strategies are being continuously developed, the complexity of the biofilm community structure and the surrounding environment means that there is unlikely to be a single solution that will minimize biofilms on all environmental sensors. Thus, antibiofouling research often focuses on optimizing a specific biofilm mitigation approach for a given sensor, application, and environmental condition. While this is practical from the standpoint of a sensor developer, it makes the comparison of different mitigation strategies difficult. In this Perspective, we discuss the application of different biofouling mitigation strategies to sensing and then explore the need for the sensor community to adopt standard protocols to increase the comparability of the biofouling mitigation approaches and help sensor developers identify the most appropriate strategy for their system.