Reactive nitrogen species from free nitrous acid (FNA) cause cell lysis.

Free nitrous acid (FNA, i.e. HNO2) has been demonstrated to have broad biocidal effects on a range of microorganisms, which has direct implications for wastewater management. However, the biocidal mechanisms still remain largely unknown. This study aims to test the hypothesis that FNA will induce cell lysis via cell membrane perforations, and consequently cause cell death via proteolysis, through the use of two model organisms namely Escherichia coli K12 and Pseudomonas putida KT2440. A combination of analytical techniques that included viability assays, atomic force microscopy (AFM), protein abundance assays and proteomic analysis using Quadruple-Orbitrap™ Mass spectrometry was used to evaluate the extent of cell death and possible cell lysis mechanisms. FNA treatment at 6.09 mg/L for 24 h (conditions typically applied in applications) induced 36 ± 4.2% and 91 ± 3.5% cell death/lysis of E. coli and P. putida, respectively. AFM showed that the lysis of cells was observed via perforations in the cell membrane; cells also appeared to shrink and become flat following FNA treatment. By introducing a reactive nitrogen species (RNS) scavenger to act as a treatment control, we further revealed that it was the nitrosative decomposition species of FNA, such as .NO that caused the cell lysis through the destruction of protein macromolecules found in the cell membrane (proteolysis). Subsequently, the RNS went on to cause the destruction of protein macromolecules within the cells. The death of these model organisms E. coli and P. putida following exposure to FNA treatment provides insights into the use of FNA as an antimicrobial agent in wastewater treatment.

Corrosion mitigation by nitrite spray on corroded concrete in a real sewer system.

Microbially influenced concrete corrosion (MICC) in sewers is caused by the activity of sulfide-oxidizing microorganisms (SOMs) on concrete surfaces, which greatly deteriorates the integrity of sewers. Surface treatment of corroded concrete by spraying chemicals is a low-cost and non-intrusive strategy. This study systematically evaluated the spray of nitrite solution in corrosion mitigation and re-establishment in a real sewer manhole. Two types of concrete were exposed at three heights within the sewer manhole for 21 months. Nitrite spray was applied at the 6th month for half of the coupons which had developed active corrosion. The corrosion development was monitored by measuring the surface pH, corrosion product composition, sulfide uptake rate, concrete corrosion loss, and the microbial community on the corrosion layer. Free nitrous acid (FNA, i.e. HNO2), formed by spraying a nitrite solution on acidic corrosion surfaces, was shown to inhibit the activity of SOMs. The nitrite spray reduced the corrosion loss of concrete at all heights by 40-90% for six months. The sulfide uptake rate of sprayed coupons was also reduced by about 35%, leading to 1-2 units higher surface pH, comparing to the control coupons. The microbial community analysis revealed a reduced abundance of SOMs on nitrite sprayed coupons. The long-term monitoring also showed that the corrosion mitigation effect became negligible in 15 months after the spray. The results consistently demonstrated the effectiveness of nitrite spray on the MICC mitigation and identified the re-application frequencies for full scale applications.

Evaluation of continuous and intermittent trickling strategies for the removal of hydrogen sulfide in a biotrickling filter.

Biotrickling filter (BTF) is a widely applied bioreactor for odour abatement in sewer networks. The trickling strategy is vital for maintaining a sound operation of BTF. This study employed a lab-scale BTF packed with granular activated carbon at a short empty bed residence time of 6 s and pH 1-2 to evaluate different trickling strategies, i.e., continuous trickling (different velocities) and intermittent trickling (different trickling intervals), in terms of the removal of hydrogen sulfide (H2S), bed pressure drop, H2S oxidation products and microbial community. The H2S removal performance decreased with the trickling velocity (∼3.6 m/h) in BTF. In addition, three intermittent trickling strategies, i.e., 10-min trickling per 24 h, 8 h, and 2 h, were investigated. The H2S elimination capacity deteriorated after about 2 weeks under both 10-min trickling per 24 h and 8 h. For both intermittent (10-min trickling per 2 h) and continuous trickling, the BTF exhibited nearly 100 % H2S removal for inlet H2S concentrations<100 ppmv, but intermittent BTF showed better removal performance than continuous trickling when inlet H2S increased to 120-190 ppmv. Furthermore, the bed pressure drops were 333 and 3888 Pa/m for non-trickling and trickling periods, respectively, which makes intermittent BTF save 83 % energy consumption of the blower compared with continuous tirckling. However, intermittent BTF exhibited transient H2S breakthrough (<1 ppmv) during trickling periods. Moreover, elemental sulfur and sulfate were major products of H2S oxidation and Acidithiobacillus was the dominant genus in both intermittent and continuous trickling BTF. A mathematical model was calibrated for the intermittent BTF and a sensitivity analysis was performed on the model. It shows mass transfer parameters determine the H2S removal. Overall, intermittent trickling strategy is promising for improving odour abatement performance and reducing the operating cost of the BTF.

Characterizing the premise plumbing microbiome in both water and biofilms of a 50-year-old building.

The premise plumbing portion of drinking water distribution systems (DWDS) has several characteristics that may favor microbial growth in the form of biofilms. These microbial communities are implicated as infectious sources for the spread of opportunistic waterborne pathogens by supporting their complex ecology and transmission through DWDS outlets to susceptible individuals. However, there is limited understanding of the drinking water biofilms in real premise plumbing networks due to challenges with accessibility. Using a combination of culture-dependent and culture-independent approaches, this study comprehensively characterized the premise plumbing microbiome of a 50-year-old university building, inclusive of water and biofilm samples. Microbial diversity in the water samples were more taxonomically diverse in comparison to the mature drinking water biofilms, which were dominated with biofilm-formers and opportunistic pathogens, such as Mycobacterium spp. A model opportunistic pathogen, Legionella spp., was only detectable in water samples using quantitative PCR but could not be detected in any of the drinking water biofilms using either qPCR or culture-dependent approaches, highlighting the limitations of detection methods in these environments. This study presents preliminary findings on the microbial dynamics and complexity in premise plumbing networks, which may support public health management and the development of strategies to eliminate microbial risks to human health.

Chlorine disinfection facilitates natural transformation through ROS-mediated oxidative stress.

The bacterial infection that involves antimicrobial resistance is a rising global threat to public health. Chlorine-based water disinfection processes can inactivate antibiotic resistant bacteria. However, at the same time, these processes may cause the release of antibiotic resistance genes into the water as free DNA, and consequently increase the risk to disseminate antibiotic resistance via natural transformation. Presently, little is known about the contribution of residual chlorine affecting the transformation of extracellular antibiotic resistance genes (ARGs). This study investigates whether chloramine and free chlorine promote the transformation of ARGs and how this may occur. We reveal that both chloramine and free chlorine, at practically relevant concentrations, significantly stimulated the transformation of plasmid-encoded ARGs by the recipient Acinetobacter baylyi ADP1, by up to a 10-fold increase. The underlying mechanisms underpinning the increased transformations were revealed. Disinfectant exposure induced a series of cell responses, including increased levels of reactive oxygen species (ROS), bacterial membrane damage, ROS-mediated DNA damage, and increased stress response. These effects thus culminated in the enhanced transformation of ARGs. This promoted transformation was observed when exposing disinfectant-pretreated A. baylyi to free plasmid. In contrast, after pretreating free plasmid with disinfectants, the transformation of ARGs decreased due to the damage of plasmid integrity. These findings provide important insight on the roles of disinfectants affecting the horizontal transfer of ARGs, which could be crucial in the management of antibiotic resistance in our water systems.

Biotrickling filter for the removal of volatile sulfur compounds from sewers: A review.

Volatile sulfur compounds (VSCs) were identified as the dominant priority odorants emitted from sewers, including hydrogen sulfide (H2S), methyl mercaptan (MM), dimethyl disulfide (DMDS) and dimethyl sulfide (DMS). Biotrickling filter (BTF) is a widely-applied technology for odour abatement in sewers because of its relatively low operating cost and efficient H2S removal. The authors review the mechanisms and performance of BTF for the removal of these four VSCs, and discuss the key influencing factors including of empty bed residence time (EBRT), pH, temperature, nutrients, water content, trickling operation and packing materials. Besides, measures to improve the VSCs removal in BTF are proposed in the context of key influencing factors. Finally, the review assesses the new challenges of BTF for sewer emissions treatment, namely with respect to the performance of BTF for greenhouse gases (GHG) treatment.

Non-antibiotic pharmaceuticals promote the transmission of multidrug resistance plasmids through intra- and intergenera conjugation.

Antibiotic resistance is a global threat to public health. The use of antibiotics at sub-inhibitory concentrations has been recognized as an important factor in disseminating antibiotic resistance via horizontal gene transfer. Although non-antibiotic, human-targeted pharmaceuticals are widely used by society (95% of the pharmaceuticals market), the potential contribution to the spread of antibiotic resistance is not clear. Here, we report that commonly consumed, non-antibiotic pharmaceuticals, including nonsteroidal anti-inflammatories (ibuprofen, naproxen, diclofenac), a lipid-lowering drug (gemfibrozil), and a ß-blocker (propranolol), at clinically and environmentally relevant concentrations, significantly accelerated the dissemination of antibiotic resistance via plasmid-borne bacterial conjugation. Various indicators were used to study the bacterial response to these drugs, including monitoring reactive oxygen species (ROS) and cell membrane permeability by flow cytometry, cell arrangement, and whole-genome RNA and protein sequencing. Enhanced conjugation correlated well with increased production of ROS and cell membrane permeability. Additionally, these non-antibiotic pharmaceuticals induced responses similar to those detected when bacteria are exposed to antibiotics, such as inducing the SOS response and enhancing efflux pumps. The findings advance understanding of the transfer of antibiotic resistance genes, emphasizing the concern that non-antibiotic, human-targeted pharmaceuticals enhance the spread of antibiotic resistance among bacterial populations.

Nonnutritive sweeteners can promote the dissemination of antibiotic resistance through conjugative gene transfer.

Antimicrobial resistance (AMR) poses a worldwide threat to human health and biosecurity. The spread of antibiotic resistance genes (ARGs) via conjugative plasmid transfer is a major contributor to the evolution of this resistance. Although permitted as safe food additives, compounds such as saccharine, sucralose, aspartame, and acesulfame potassium that are commonly used as nonnutritive sweeteners have recently been associated with shifts in the gut microbiota similar to those caused by antibiotics. As antibiotics can promote the spread of antibiotic resistance genes (ARGs), we hypothesize that these nonnutritive sweeteners could have a similar effect. Here, we demonstrate for the first time that saccharine, sucralose, aspartame, and acesulfame potassium could promote plasmid-mediated conjugative transfer in three established conjugation models between the same and different phylogenetic strains. The real-time dynamic conjugation process was visualized at the single-cell level. Bacteria exposed to the tested compounds exhibited increased reactive oxygen species (ROS) production, the SOS response, and gene transfer. In addition, cell membrane permeability increased in both parental bacteria under exposure to the tested compounds. The expression of genes involved in ROS detoxification, the SOS response, and cell membrane permeability was significantly upregulated under sweetener treatment. In conclusion, exposure to nonnutritive sweeteners enhances conjugation in bacteria. Our findings provide insight into AMR spread and indicate the potential risk associated with the presence of nonnutritive sweeteners.

Structural changes in model compounds of sludge extracellular polymeric substances caused by exposure to free nitrous acid.

Previous studies demonstrate that free nitrous acid (FNA i.e. HNO2) detaches sewer biofilms, breaks down flocs of waste activated sludge (WAS) and enhances biogas production from WAS. This suggests possible interactions of FNA with organic extracellular polymeric substances (EPS) that bind the cells into biofilms or sludge flocs. This study evaluates the chemical interactions and reaction mechanisms between FNA and molecules representative of key EPS in biofilm and sludge flocs. Molecules chosen to represent components found in the extracellular polymeric matrix were treated with FNA at 6.09 mgN/L (NO2- = 250 mgN/L, pH = 5.0 ± 0.2, T = 22 °C) for 24 hours (conditions typically used in applications) so as to consider the hypothesized chemical interactions and the consequent reaction pathways. A number of analytical techniques were employed to measure the molecular changes in the EPS molecules including; proton (1H) nuclear magnetic resonance spectroscopy (NMR), electrospray ionisation mass spectrometry (ESI-MS) and gel permeation chromatography (GPC). The results demonstrated that FNA broke down a range of large EPS molecules including carbohydrates, protein and lipids to smaller molecules. Two mechanistic pathways have been proposed including electrophilic substitution, whereby the nitrosium ion (NO+) was the reactive electrophile, and oxidative radical reactions, through which the nitrogen radicals (.NO2, .NO) and reactive nitrogen intermediates (RNIs) (e.g. N2O3 and N2O4) formed from the decomposition of FNA became part of the reaction products. Larger, more complex organic molecules such as humic acid, required higher concentrations of FNA (6.09 mgN/L or greater) to cause molecular breakdown, whereas smaller molecules, such as calcium alginate, was broken down at lower concentrations (3.04 mgN/L). The study contributes to the understanding of the fundamental mechanisms behind the application of FNA for biofilm control and flocular sludge disintegration.

Structural Changes in Cell-Wall and Cell-Membrane Organic Materials Following Exposure to Free Nitrous Acid.

Previous studies demonstrate that free nitrous acid (FNA, i.e., HNO2) is biocidal for a range of microorganisms. The biocidal mechanisms of FNA are largely unknown. In this work, it is hypothesized that FNA will break bonds in molecules found in the cell envelope, thus causing cell lysis. Selected molecules representing components found in the cell envelope were treated with FNA at 6.09 mg N/L (NO2- = 250 mg N/L, pH 5.0) for 24 h (conditions typically used in applications) to evaluate the hypothesized chemical interactions. Molecular changes were observed using analytical techniques including proton (1H) nuclear magnetic resonance spectroscopy (NMR) and electrospray ionization mass spectrometry (ESI-MS). It was found that FNA broke down a range of cell envelope molecules. The spectral data demonstrated that the FNA reactions proceeded via two general pathways. One consisted of electrophilic substitution, whereby the nitrosonium ion (NO+) was the reactive electrophile. The other was via oxidative reactions involving nitrogen radicals (e.g., •NO2 and •NO) formed from the decomposition of FNA. We further revealed that it was HNO2 that caused the breakdown, rather than the exclusive action of the acid (H+) or nitrite (NO2-) counterparts. The fragmentation of these representative cell envelope molecules provides insight into the biocidal effects of FNA on microorganisms.

Non-antibiotic pharmaceuticals enhance the transmission of exogenous antibiotic resistance genes through bacterial transformation.

Antibiotic resistance is a serious global threat for public health. Considering the high abundance of cell-free DNA encoding antibiotic resistance genes (ARGs) in both clinical and environmental settings, natural transformation is an important horizontal gene transfer pathway to transmit antibiotic resistance. It is acknowledged that antibiotics are key drivers for disseminating antibiotic resistance, yet the contributions of non-antibiotic pharmaceuticals on transformation of ARGs are overlooked. In this study, we report that some commonly consumed non-antibiotic pharmaceuticals, at clinically and environmentally relevant concentrations, significantly facilitated the spread of antibiotic resistance through the uptake of exogenous ARGs. This included nonsteroidal anti-inflammatories, ibuprofen, naproxen, diclofenac, the lipid-lowering drug, gemfibrozil, and the ß-blocker propranolol. Based on the results of flow cytometry, whole-genome RNA sequencing and proteomic analysis, the enhanced transformation of ARGs was affiliated with promoted bacterial competence, enhanced stress levels, over-produced reactive oxygen species and increased cell membrane permeability. In addition, a mathematical model was proposed and calibrated to predict the dynamics of transformation during exposure to non-antibiotic pharmaceuticals. Given the high consumption of non-antibiotic pharmaceuticals, these findings reveal new concerns regarding antibiotic resistance dissemination exacerbated by non-antibiotic pharmaceuticals.

Efficient inactivation of antibiotic resistant bacteria and antibiotic resistance genes by photo-Fenton process under visible LED light and neutral pH.

Antibiotic resistance has been recognized as a major threat to public health worldwide. Inactivation of antibiotic resistant bacteria (ARB) and degradation of antibiotic resistance genes (ARGs) are critical to prevent the spread of antibiotic resistance in the environment. Conventional disinfection processes are effective to inactivate water-borne pathogens, yet they are unable to completely eliminate the antibiotic resistance risk. This study explored the potential of the photo-Fenton process to inactivate ARB, and to degrade both extracellular and intracellular ARGs (e-ARGs and i-ARGs, respectively). Using Escherichia coli DH5α with two plasmid-encoded ARGs (tetA and blaTEM-1) as a model ARB, a 6.17 log ARB removal was achieved within 30 min of applying photo-Fenton under visible LED and neutral pH conditions. In addition, no ARB regrowth occurred after 48-h, demonstrating that this process is very effective to induce permanent disinfection on ARB. The photo-Fenton process was validated under various water matrices, including ultrapure water (UPW), simulated wastewater (SWW) and phosphate buffer (PBS). The higher inactivation efficiency was observed in SWW as compared to other matrices. The photo-Fenton process also caused a 6.75 to 8.56-log reduction in eARGs based on quantitative real-time PCR of both short- and long amplicons. Atomic force microscopy (AFM) further confirmed that the extracellular DNA was sheared into short DNA fragments, thus eliminating the risk of the transmission of antibiotic resistance. As compared with e-ARGs, a higher dosage of Fenton reagent was required to damage i-ARGs. In addition, the tetA gene was more easily degraded than the blaTEM-1 gene. Collectively, our results demonstrate the photo-Fenton process is a promising technology for disinfecting water to prevent the spread of antibiotic resistance.

Adaptive Evolution of Geobacter sulfurreducens in Coculture with Pseudomonas aeruginosa.

Interactions between microorganisms in mixed communities are highly complex, being either syntrophic, neutral, predatory, or competitive. Evolutionary changes can occur in the interaction dynamics between community members as they adapt to coexistence. Here, we report that the syntrophic interaction between Geobacter sulfurreducens and Pseudomonas aeruginosa coculture change in their dynamics over evolutionary time. Specifically, Geobacter sp. dominance increases with adaptation within the cocultures, as determined through quantitative PCR and fluorescence in situ hybridization. This suggests a transition from syntrophy to competition and demonstrates the rapid adaptive capacity of Geobacter spp. to dominate in cocultures with P. aeruginosa Early in coculture establishment, two single-nucleotide variants in the G. sulfurreducensfabI and tetR genes emerged that were strongly selected for throughout coculture evolution with P. aeruginosa phenazine wild-type and phenazine-deficient mutants. Sequential window acquisition of all theoretical spectra-mass spectrometry (SWATH-MS) proteomics revealed that the tetR variant cooccurred with the upregulation of an adenylate cyclase transporter, CyaE, and a resistance-nodulation-division (RND) efflux pump notably known for antibiotic efflux. To determine whether antibiotic production was driving the increased expression of the multidrug efflux pump, we tested Pseudomonas-derived phenazine-1-carboxylic acid (PHZ-1-CA) for its potential to inhibit Geobacter growth and drive selection of the tetR and fabI genetic variants. Despite its inhibitory properties, PHZ-1-CA did not drive variant selection, indicating that other antibiotics may drive overexpression of the efflux pump and CyaE or that a novel role exists for these proteins in the context of this interaction.IMPORTANCEGeobacter and Pseudomonas spp. cohabit many of the same environments, where Geobacter spp. often dominate. Both bacteria are capable of extracellular electron transfer (EET) and play important roles in biogeochemical cycling. Although they recently in 2017 were demonstrated to undergo direct interspecies electron transfer (DIET) with one another, the genetic evolution of this syntrophic interaction has not been examined. Here, we use whole-genome sequencing of the cocultures before and after adaptive evolution to determine whether genetic selection is occurring. We also probe their interaction on a temporal level and determine whether their interaction dynamics change over the course of adaptive evolution. This study brings to light the multifaceted nature of interactions between just two microorganisms within a controlled environment and will aid in improving metabolic models of microbial communities comprising these two bacteria.

Correction to: Molecular diversity of arbuscular mycorrhizal fungal communities across the gradient of alkaline Fe ore tailings, revegetated waste rock to natural soil sites.

The authors thank the Australian Centre for Ecogenomics in the University of Queensland for conducting Illumina sequencing.

Triclosan at environmental concentrations can enhance the spread of extracellular antibiotic resistance genes through transformation.

The dissemination of antibiotic resistance mediated by horizontal transfer of antibiotic resistance genes (ARGs) is exacerbating the global antibiotic crisis. Currently, little is known about whether non-antibiotic, anti-microbial (NAAM) chemicals are associated with the dissemination of ARGs in the environment. In this study, we aimed to evaluate whether a ubiquitous NAAM chemical, triclosan (TCS), is able to promote the transformation of plasmid-borne antibiotic resistance genes (ARGs). By using the plasmid pUC19 carrying ampicillin resistance genes as the extracellular ARG and a model microorganism Escherichia coli DH5ɑ as the recipient, we found that TCS at environmentally detected concentrations (0.2 µg/L to 20 µg/L) significantly enhanced the transformation of plasmid-borne ARGs into E. coli DH5ɑ for up to 1.4-fold. The combination of phenotypic experiments, genome-wide RNA sequencing and proteomic analyses revealed that TCS exposure stimulated the reactive oxygen species (ROS) production for 1.3- to 1.5-fold, induced bacterial membrane damage and up-regulated the translation of outer membrane porin. Moreover, general secretion system Sec (1.4-fold), twin arginine translocation system Tat (1.2-fold) and type IV pilus secretion systems (2.5-fold) were enhanced by TCS, which might contribute to the DNA searching/capture by pilus. Together, TCS might increase the transformation frequency of ARGs into E. coli DH5ɑ by ROS over-production, damaging cell membrane barrier, mediating the pilus capture of plasmid and the translocation of plasmid via cell membrane channels. This study reports that TCS could accelerate the transformation of extracellular ARGs to competent bacteria at environmentally relevant concentrations. The findings advance our understanding of the fate of ARGs in ecosystems and call for risk assessments of NAAM chemicals on disseminating antibiotic resistance.

Molecular diversity of arbuscular mycorrhizal fungal communities across the gradient of alkaline Fe ore tailings, revegetated waste rock to natural soil sites.

Arbuscular mycorrhizal (AM) fungi are important to the establishment of native vegetation for mined land rehabilitation, particularly in semi-arid and infertile landscapes. However, the information has been scarce about the colonization of AM fungal community in alkaline magnetite Fe ore tailing sites (without toxic metal (loid) contamination). The present study has characterized the diversity of AM fungi across typical domains of a magnetite Fe ore mine located in 200 km south-east of Geraldton, Western Australia, by adopting high throughput Illumina Miseq sequencing. The investigated domains included two tailing sites without top soil covering (T1 and T2), a rehabilitated area of tailings with top soil covering (R1), a revegetated waste rock area (R2), and two native undisturbed soil sites (S1 and S2). The results indicated that the T1/T2 sites had different AM fungal community structure, compared with R1/R2 and S1/S2 sites. The dominant families were Glomeraceae, Claroideoglomeraceae, Archaeosporaceae, Ambisporaceae, and Paraglomeraceae, with Paraglomeraceae (more than 50%) as the most abundant in the T1/T2 and R1/R2 sites. At genus level, Ambispora spp. and Archaeospora spp. were rich in T1/T2 sites (> 10%), while Glomus spp. were preferably dominant in S1/S2 sites (> 10%). Furthermore, amorphous Fe and available P were found to explain the variations associated with AM fungal community composition, particularly the abundance of Archaeosporaceae and Glomeraceae. The study revealed the AM fungal community composition shift across the gradient of Fe ore mine sites, as well as the effects of revegetation on AM fungal community development. The findings indicate the possible restoration of AM fungal community in the tailings undergoing revegetation, and potential adoption of indigenous AM fungi to rapid phytostabilization of the Fe ore tailings under semi-arid climatic conditions.

Increased Resistance of Nitrite-Admixed Concrete to Microbially Induced Corrosion in Real Sewers.

Microbially induced concrete corrosion is a major deterioration process in sewers, causing a huge economic burden, and improved mitigating technologies are required. This study reports a novel and promising effective solution to attenuate the corrosion in sewers using calcium nitrite-admixed concrete. This strategy aims to suppress the development and activity of corrosion-inducing microorganisms with the antimicrobial free nitrous acid, which is generated in situ from calcium nitrite that is added to the concrete. Concrete coupons with calcium nitrite as an admixture were exposed in a sewer manhole, together with control coupons that had no nitrite admixture, for 18 months. The corrosion process was monitored by measuring the surface pH, corrosion product composition, concrete corrosion loss, and the microbial community on the corrosion layer. During the exposure, the corrosion loss of the admixed concrete coupons was 30% lower than that of the control coupons. The sulfide uptake rate of the admixed concrete was also 30% lower, leading to a higher surface pH (0.5-0.6 unit), in comparison to that of the control coupons. A negative correlation between the calcium nitrite admixture in concrete and the abundance of sulfide-oxidizing microorganisms was determined by DNA sequencing. The results obtained in this field study demonstrated that this novel use of calcium nitrite as an admixture in concrete is a promising strategy to mitigate the microbially induced corrosion in sewers.

Improving wastewater management using free nitrous acid (FNA).

Free nitrous acid (FNA), the protonated form of nitrite, has historically been an unwanted substance in wastewater systems due to its inhibition on a wide range of microorganisms. However, in recent years, advanced understanding of FNA inhibitory and biocidal effects on microorganisms has led to the development of a series of FNA-based applications that improve wastewater management practices. FNA has been used in sewer systems to control sewer corrosion and odor; in wastewater treatment to achieve carbon and energy efficient nitrogen removal; in sludge management to improve the sludge reduction and energy recovery; in membrane systems to address membrane fouling; and in wastewater algae systems to facilitate algae harvesting. This paper aims to comprehensively and critically review the current status of FNA-based applications in improving wastewater management. The underlying mechanisms of FNA inhibitory and biocidal effects are also reviewed and discussed. Knowledge gaps and current limitations of the FNA-based applications are identified; and perspectives on the development of FNA-based applications are discussed. We conclude that the FNA-based technologies have great potential for enhancing the performance of wastewater systems; however, further development and demonstration at larger scales are still required for their wider applications.