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Aybar, M., Perez-Calleja, P., Li, M., Pavissich, J. P., & Nerenberg, R. (2019). Predation creates unique void layer in membrane-aerated biofilms. Water Res., 149, 232–242.
Abstract: The membrane-aerated biofilm reactor (MABR) is a novel wastewater treatment technology based on oxygen-supplying membranes. The counter diffusion of oxygen and electron donors in MABRs leads to unique behavior, and we hypothesized it also could impact predation. We used optical coherence tomography (OCT), microsensor analyses, and mathematical modeling to investigate predation in membrane-aerated biofilms (MABs). When protozoa were excluded from the inoculum, the MAB's OCT-observable void fraction was around 5%. When protozoa were included, the void fraction grew to nearly 50%, with large, continuous voids at the base of the biofilm. Real-time OCT imaging showed highly motile protozoa in the voids. MABs with protozoa and a high bulk COD (270 mg/L) only had 4% void fraction. DNA sequencing revealed a high relative abundance of amoeba in both high and low-COD MABs. Flagellates were only abundant in the low-COD MAB. Modeling also suggested a relationship between substrate concentrations, diffusion mode (co- or counter-diffusion), and bioflim void fraction. Results suggest that amoeba proliferate in the bioflim interior, especially in the aerobic zones. Voids form once COD limitation at the base of MABs allows predation rates to exceed microbial growth rates. Once formed, the voids provide a niche for motile protozoa, which expand the voids into a large, continuous gap. This increases the potential for biofilm sloughing, and may have detrimental effects on slow-growing, aerobic microorganisms such as nitrifying bacteria. (C)2018 Elsevier Ltd. All rights reserved.
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Besaury, L., Ouddane, B., Pavissich, J. P., Dubrulle-Brunaud, C., Gonzalez, B., & Quillet, L. (2012). Impact of copper on the abundance and diversity of sulfate-reducing prokaryotes in two chilean marine sediments. Mar. Pollut. Bull., 64(10), 2135–2145.
Abstract: We studied the abundance and diversity of the sulfate-reducing prokaryotes (SRPs) in two 30-cm marine chilean sediment cores, one with a long-term exposure to copper-mining residues, the other being a non-exposed reference sediment. The abundance of SRPs was quantified by qPCR of the dissimilatory sulfite reductase gene beta-subunit (dsrB) and showed that SRPs are sensitive to high copper concentrations, as the mean number of SRPs all along the contaminated sediment was two orders of magnitude lower than in the reference sediment. SRP diversity was analyzed by using the dsrB-sequences-based PCR-DGGE method and constructing gene libraries for dsrB-sequences. Surprisingly, the diversity was comparable in both sediments, with dsrB sequences belonging to Desulfobacteraceae, Syntrophobacteraceae, and Desulfobulbaceae, SRP families previously described in marine sediments, and to a deep branching dsrAB lineage. The hypothesis of the presence of horizontal transfer of copper resistance genes in the microbial population of the polluted sediment is discussed. (C) 2012 Elsevier Ltd. All rights reserved.
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Campos, J. L., Crutchik, D., Franchi, O., Pavissich, J. P., Belmonte, M., Pedrouso, A., et al. (2019). Nitrogen and Phosphorus Recovery From Anaerobically Pretreated Agro-Food Wastes: A Review. Front. Sustain. Food Syst., 2, 11 pp.
Abstract: Anaerobic digestion (AD) is commonly used for the stabilization of agro-food wastes and recovery of energy as methane. Since AD removes organic C but not nutrients (N and P), additional processes to remove them are usually applied to meet the stringent effluent criteria. However, in the past years, there was a shift from the removal to the recovery of nutrients as a result of increasing concerns regarding limited natural resources and the importance given to the sustainable treatment technologies. Recovering N and P from anaerobically pretreated agro-food wastes as easily transportable and marketable products has gained increasing importance to meet both regulatory requirements and increase revenue. For this reason, this review paper gives a critical comparison of the available and emerging technologies for N and P recovery from AD residues.
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Campos, J. L., Dumais, J., Pavissich, J. P., Franchi, O., Crutchik, D., Belmonte, M., et al. (2019). Predicting Accumulation of Intermediate Compounds in Nitrification and Autotrophic Denitrification Processes: A Chemical Approach. Biomed Res. Int., 2019, 9 pp.
Abstract: Nitrification and sulfur-based autotrophic denitrification processes can be used to remove ammonia from wastewater in an economical way. However, under certain operational conditions, these processes accumulate intermediate compounds, such as elemental sulphur, nitrite, and nitrous oxide, that are noxious for the environment. In order to predict the generation of these compounds, an analysis based on the Gibbs free energy of the possible reactions and on the oxidative capacity of the bulk liquid was done on case study systems. Results indicate that the Gibbs free energy is not a useful parameter to predict the generation of intermediate products in nitrification and autotrophic denitrification processes. Nevertheless, we show that the specific productions of nitrous oxide during nitrification, and of elemental sulphur and nitrite during autotrophic denitrification, are well related to the oxidative capacity of the bulk liquid.
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Cofre, C., Campos, J. L., Valenzuela-Heredia, D., Pavissich, J. P., Camus, N., Belmonte, M., et al. (2018). Novel system configuration with activated sludge like-geometry to develop aerobic granular biomass under continuous flow. Bioresour. Technol., 267, 778–781.
Abstract: A novel continuous flow system with “flat geometry” composed by two completely mixed aerobic tanks in series and a settler was used to promote the formation of aerobic granular sludge. Making similarities of this system with a typical sequencing batch reactor (SBR), for aerobic granules cultivation, the value of the tank 1/tank 2 vol ratio and the biomass recirculation rate would correspond with the feast/famine length ratio and the length of the operational cycle, respectively, while the settler upflow liquid velocity imposed would be related to the settling time. From the three experiments performed the best results were obtained when the tank 1/tank 2 vol ratio was of 0.28, the sludge recycling ratio of 0.25 and the settler upflow velocity of 2.5 m/h. At these conditions the aggregates had settling velocities between 29 and 113 m/h, sludge volume index at 10 min (SVI10) of 70 mL/g TSS and diameters between 1.0 and 5.0 mm.
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De la Iglesia, R., Valenzuela-Heredia, D., Pavissich, J. P., Freyhoffer, S., Andrade, S., Correa, J. A., et al. (2010). Novel polymerase chain reaction primers for the specific detection of bacterial copper P-type ATPases gene sequences in environmental isolates and metagenomic DNA. Lett. Appl. Microbiol., 50(6), 552–562.
Abstract: Aims: In the last decades, the worldwide increase in copper wastes release by industrial activities like mining has driven environmental metal contents to toxic levels. For this reason, the study of the biological copper-resistance mechanisms in natural environments is important. Therefore, an appropriate molecular tool for the detection and tracking of copper-resistance genes was developed. Methods and Results: In this work, we designed a PCR primer pair to specifically detect copper P-type ATPases gene sequences. These PCR primers were tested in bacterial isolates and metagenomic DNA from intertidal marine environments impacted by copper pollution. As well, T-RFLP fingerprinting of these gene sequences was used to compare the genetic composition of such genes in microbial communities, in normal and copper-polluted coastal environments. New copper P-type ATPases gene sequences were found, and a high degree of change in the genetic composition because of copper exposure was also determined. Conclusions: This PCR based method is useful to track bacterial copper-resistance gene sequences in the environment. Significance and Impact of the Study: This study is the first to report the design and use of a PCR primer pair as a molecular marker to track bacterial copper-resistance determinants, providing an excellent tool for long-term analysis of environmental communities exposed to metal pollution.
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Franchi, O., Alvarez, M. I., Pavissich, J. P., Belmonte, M., Pedrouso, A., del Rio, A. V., et al. (2024). Operational variables and microbial community dynamics affect granulation stability in continuous flow aerobic granular sludge reactors. J. Water Process Eng., 59, 104951.
Abstract: Retrofitting wastewater treatment plants with continuous aerobic granular sludge reactors is a promising alternative to enhance treatment capacities and reduce footprint. This study investigates the main variables influencing granulation and microbial dynamics in two reactor configurations (25 L): stirred tanks in series (R1) and a plug-flow-like system (R2). Granule formation was achieved by increasing the organic loading rate (OLR) from 0.7 to 4.1 kg COD/(m3 & sdot;d) and the up-flow velocity in the biomass selector from 1.4 to 6.9 m/h. However, irreversible granule destabilization occurred at day 68 for R1 and day 108 for R2. Principal component analysis and examination of food-to-microorganisms (F/M) ratio medians identified the F/M ratio as the primary variable associated with instability. Microbial analysis revealed that a high F/M ratio induced significant increases in the abundance of specific genera such as Arcobacter, Cloacibacterium, Rikenella, Aquaspirillum and Sphaerotillus, whose overgrowth may negatively impact granule stability. Based on these findings, maximum F/M ratio thresholds were obtained to establish operational conditions allowing the maintenance of stable aerobic granules on continuous flow reactor configurations.
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McFadden, M., Loconsole, J., Schockling, A., Nerenberg, R., & Pavissich, J. P. (2017). Comparing peracetic acid and hypochlorite for disinfection of combined sewer overflows: Effects of suspended-solids and pH. Sci. Total Environ., 599, 533–539.
Abstract: Peracetic acid (PM) is an alternative disinfectant that may be effective for combined sewer overflow (CSO) disinfection, but little is known about the effect of particle size on PM disinfection efficiency. In this work, PM and hypochlorite were compared as disinfectants, with a focus on the effect of wastewater particles. Inactivation experiments were conducted on suspended cultures of Escherichia coli and wastewater suspended solids. Tested size fractions included particle diameters <10 μm, <100 μm, and raw wastewater. Chlorine disinfection efficiency decreased with increasing solids size. However, solids size had little effect on PM disinfection. The PM disinfection efficiency decreased at pH values above 7.5. Live/dead staining revealed that PM disinfection leaves most cells in a viable but non-culturable condition. Fourier transform infrared spectroscopy (FTIR) analyses suggests that PAA and hypochlorite may inactivate E. coli bacteria by similar mechanisms. (C) 2017 Elsevier B.V. All rights reserved.
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Pavissich, J. P., Li, M., & Nerenberg, R. (2021). Spatial distribution of mechanical properties in Pseudomonas aeruginosa biofilms, and their potential impacts on biofilm deformation. Biotechnol. Bioeng., 118(4), 1564–1575.
Abstract: The mechanical properties of biofilms can be used to predict biofilm deformation under external forces, for example under fluid flow. We used magnetic tweezers to spatially map the compliance of Pseudomonas aeruginosa biofilms at the microscale, then applied modeling to assess its effects on biofilm deformation. Biofilms were grown in capillary flow cells with Reynolds numbers (Re) ranging from 0.28 to 13.9, bulk dissolved oxygen (DO) concentrations from 1 mg/L to 8 mg/L, and bulk calcium ion (Ca2+) concentrations of 0 and 100 mg CaCl2/L. Higher Re numbers resulted in more uniform biofilm morphologies. The biofilm was stiffer at the center of the flow cell than near the walls. Lower bulk DO led to more stratified biofilms. Higher Ca2+ concentrations led to increased stiffness and more uniform mechanical properties. Using the experimental mechanical properties, fluid‐structure interaction models predicted up to 64% greater deformation for heterogeneous biofilms, compared to a homogeneous biofilms with the same average properties. However, the deviation depended on the biofilm morphology and flow regime. Our results show significant spatial mechanical variability exists at the microscale, and that this variability can potentially affect biofilm deformation. The average biofilm mechanical properties, provided in many studies, should be used with caution when predicting biofilm deformation.
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Pavissich, J. P., Camus, N., Campos, J. L., Franchi, O., Pedrouso, A., Carrera, P., et al. (2021). Monitoring the stability of aerobic granular sludge using fractal dimension analysis. Environ. Sci-Wat Res., 7(4), 706–713.
Abstract: Cyclic episodes of granules formation and disintegration took place in two lab-scale aerobic granular sludge sequencing batch reactors, one fed with synthetic wastewater (COD: 0.6 g L-1 and NH4+-N: 0.06 g L-1) and operated at a constant organic loading rate (2.5 g COD per L d), and the other fed with real wastewater (soluble COD: 0.27-1.37 and NH4+-N: 0.02-0.16 g L-1) and with a variable loading rate (between 1.1 and 5.5 g CODsoluble per L d). The sludge volume index, density and diameter (mean value and relative standard deviation) of the granular biomass showed great fluctuations, without any clear tendency during the operational period. However, changes in granules fractal dimension values (both mean and relative standard deviation) matched with the formation and disintegration dynamics of the granular biomass. Statistical data analysis showed that the relative standard deviation of the granules fractal dimension could be a useful parameter for monitoring the granules status. Indeed, an increase of its value during the maturation or steady-state granulation stages is an early warning of disintegration episodes. A control strategy to maintain granules integrity based on this parameter is proposed.
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Pavissich, J. P., Silva, M., & Gonzalez, B. (2010). Sulfate Reduction, Molecular Diversity, And Copper Amendment Effects In Bacterial Communities Enriched From Sediments Exposed To Copper Mining Residues. Environ. Toxicol. Chem., 29(2), 256–264.
Abstract: Sulfate-reducing bacterial communities from coastal sediments with a long-term exposure to copper (Cu)-mining residues were studied in lactate enrichments. The toxicity of excess copper may affect sulfate-reducing bacterial communities. Sulfate reduction was monitored by sulfate and organic acid measurements. Molecular diversity was analyzed by 16S rRNA, dissimilatory sulfate reduction dsrAB, and Cu translocating phospho-type adenosine triphosphatases (P-ATPases) cop-like gene sequence profiling. The influence of Cu amendment was tested in these enrichments. Results showed fast sulfate reduction mostly coupled to incomplete organic carbon oxidation and partial sulfate reduction inhibition due to copper amendment. The 16S rRNA clonal libraries analysis indicated that delta- and gamma-Proteobacteria and Cytophaga-Flexibacter-Bacteroides dominated the enrichments. The dsrAB libraries revealed the presence of Desulfobacteraceae and Desulfovibrionaceae families-related sequences. Copper produced significant shifts (i.e., a decrease in the relative abundance of sulfate-reducing microorganisms) in the enriched bacterial community structure as determined by terminal-restriction fragment length polymorphism (T-RFLP) profiling and multivariate analyses. Clonal libraries of cop-like sequences showed low richness in the enriched microbial communities, and a strong effect of copper on its relative abundance. Novel Cu-P-IB-ATPase sequences encoding Cu resistance were detected. The present study indicates that Cu does not significantly affect sulfate reduction and genetic diversity of taxonomic and dissimilatory sulfate-reduction molecular markers. However, the diversity of Cu resistance genetic determinants was strongly modified by this toxic metal. Environ. Toxicol. Chem. 2010;29:256-264. (C) 2009 SETAC
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Pavissich, J. P., Vargas, I. T., Gonzalez, B., Pasten, P. A., & Pizarro, G. E. (2010). Culture dependent and independent analyses of bacterial communities involved in copper plumbing corrosion. J. Appl. Microbiol., 109(3), 771–782.
Abstract: Aims: This study used culture-dependent and culture-independent approaches to characterize bacterial communities in copper plumbing corrosion and to assess biofilm formation and copper resistance of heterotrophic bacteria isolated from copper pipes. Methods and Results: Water and copper pipes were collected from a cold-water household distribution system affected by 'blue water' corrosion and presenting biofilm formation. Corrosion-promoting ageing experiments were performed with conditioned unused copper pipes filled with unfiltered and filtered sampled water as nonsterile and sterile treatments, respectively. During 8 weeks, stagnant water within the pipes was replaced with aerated fresh water every 2 or 3 days. Total copper and pH were determined in sampled water, and copper pipe coupons were cut for microscopic analyses. Biofilms were extracted from field and laboratory pipes, and total DNA was isolated. Bacterial communities' composition was analysed by terminal restriction fragment length polymorphism (T-RFLP) and clonal libraries of 16S rRNA genes. Heterotrophic bacterial isolates were obtained from water and biofilm extracts and characterized in terms of biofilm formation capacity and copper minimum inhibitory concentration. The results indicated that copper concentration in stagnant water from nonsterile treatments was much higher than in sterile treatments and corrosion by-products structure in coupon surfaces was different. Multivariate analysis of T-RFLP profiles and clone sequencing showed significant dissimilarity between field and laboratory biofilm communities, and a low richness and the dominant presence of Gamma- and Betaproteobacteria in both cases. Several bacterial isolates formed biofilm and tolerated high copper concentrations. Conclusions: The study demonstrates microbially influenced corrosion (MIC) in copper plumbing. Gamma- and Betaproteobacteria dominated the corroded copper piping bacterial community, whose ability to form biofilms may be important for bacterial corrosion promotion and survival in MIC events. Significance and Impact of the Study: The characterization of micro-organisms that influence copper plumbing corrosion has significant implications for distribution system management and copper corrosion control.
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Read-Daily, B. L., Sabba, F., Pavissich, J. P., & Nerenberg, R. (2016). Kinetics of nitrous oxide (N2O) formation and reduction by Paracoccus pantotrophus. AMB Express, 6, 7 pp.
Abstract: Nitrous oxide (N2O) is a powerful greenhouse gas emitted from wastewater treatment, as well as natural systems, as a result of biological nitrification and denitrification. While denitrifying bacteria can be a significant source of N2O, they can also reduce N2O to N-2. More information on the kinetics of N2O formation and reduction by denitrifying bacteria is needed to predict and quantify their impact on N2O emissions. In this study, kinetic parameters were determined for Paracoccus pantotrophus, a common denitrifying bacterium. Parameters included the maximum specific reduction rates, (q) over cap, growth rates, (mu) over cap, and yields, Y, for reduction of NO3- (nitrate) to nitrite (N2O-), N2O- to N2O, and N2O to N-2, with acetate as the electron donor. The (q) over cap values were 2.9 gN gCOD(-1) d(-1) for NNO3- to NO2-, 1.4 gN gCOD(-1) d(-1) for N2O-to N2O, and 5.3 gN gCOD(-1) d(-1) for N2O to N-2. The (mu) over cap values were 2.7, 0.93, and 1.5 d(-1), respectively. When N2O and NO3- were added concurrently, the apparent (extant) kinetics, (q) over cap (app), assuming reduction to N-2, were 6.3 gCOD gCOD(-1) d(-1), compared to 5.4 gCOD gCOD(-1) d(-1) for NO3- as the sole added acceptor. The (mu) over cap (app) was 1.6 d(-1), compared to 2.5 d(-1) for NO3- alone. These results suggest that NO3- and N2O were reduced concurrently. Based on this research, denitrifying bacteria like P. pantotrophus may serve as a significant sink for N2O. With careful design and operation, treatment plants can use denitrifying bacteria to minimize N2O emissions.
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Tierra, G., Pavissich, J. P., Nerenberg, R., Xu, Z. L., & Alber, M. S. (2015). Multicomponent model of deformation and detachment of a biofilm under fluid flow. J. R. Soc. Interface, 12(106), 13 pp.
Abstract: A novel biofilm model is described which systemically couples bacteria, extracellular polymeric substances (EPS) and solvent phases in biofilm. This enables the study of contributions of rheology of individual phases to deformation of biofilm in response to fluid flow as well as interactions between different phases. The model, which is based on first and second laws of thermodynamics, is derived using an energetic variational approach and phase-field method. Phase-field coupling is used to model structural changes of a biofilm. A newly developed unconditionally energy-stable numerical splitting scheme is implemented for computing the numerical solution of the model efficiently. Model simulations predict biofilm cohesive failure for the flow velocity between O(10(-3)) and O(10(-2))ms(-1) which is consistent with experiments. Simulations predict biofilm deformation resulting in the formation of streamers for EPS exhibiting a viscous-dominated mechanical response and the viscosity of EPS being less than O(10) kgm(-1) s(-1). Higher EPS viscosity provides biofilm with greater resistance to deformation and to removal by the flow. Moreover, simulations show that higher EPS elasticity yields the formation of streamers with complex geometries that are more prone to detachment. These model predictions are shown to be in qualitative agreement with experimental observations.
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Vargas, I. T., Fischer, D. A., Alsina, M. A., Pavissich, J. P., Pasten, P. A., & Pizarro, G. E. (2017). Copper Corrosion and Biocorrosion Events in Premise Plumbing. Materials, 10(9), 30 pp.
Abstract: Corrosion of copper pipes may release high amounts of copper into the water, exceeding the maximum concentration of copper for drinking water standards. Typically, the events with the highest release of copper into drinking water are related to the presence of biofilms. This article reviews this phenomenon, focusing on copper ingestion and its health impacts, the physicochemical mechanisms and the microbial involvement on copper release, the techniques used to describe and understand this phenomenon, and the hydrodynamic effects. A conceptual model is proposed and the mathematical models are reviewed.
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Walker, P., Nerenberg, R., Pizarro, G., Aybar, M., Pavissich, J. P., González, B., et al. (2024). Nitrate increases the capacity of an aerobic moving-bed biofilm reactor (MBBR) for winery wastewater treatment. Water Sci. Technol., Early Access.
Abstract: We used bench-scale tests and mathematical modeling to explore chemical oxygen demand (COD) removal rates in a moving-bed biofilm reactor (MBBR) for winery wastewater treatment, using either urea or nitrate as a nitrogen source. With urea addition, the COD removal fluxes ranged from 34 to 45 gCOD/m(2)-d. However, when nitrate was added, fluxes increased up to 65 gCOD/m(2)-d, twice the amount reported for aerobic biofilms for winery wastewater treatment. A one-dimensional biofilm model, calibrated with data from respirometric tests, accurately captured the experimental results. Both experimental and modelling results suggest that nitrate significantly increased MBBR capacity by stimulating COD oxidation in the deeper, oxygen-limited regions of the biofilm. Our research suggests that the addition of nitrate, or other energetic and broadly used electron acceptors, may provide a cost-effective means of covering peak COD loads in biofilm processes for winery or another industrial wastewater treatment.
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