Effect of Toxic Phthalate-Based Plasticizer on the Biodegradability of Polyhydroxyalkanoate.

While new biodegradable materials are being rapidly introduced to address plastic pollution, their end-of-life impacts remain unclear. Biodegradable plastics typically comprise a biopolymer matrix with functional additives and/or solid fillers, which may be toxic. Here, using an established method for continuous biodegradation monitoring, we investigated the impact of a commonly used plasticizer, dibutyl phthalate (DBP), on the biodegradation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) in soil. The presence of DBP delayed the initial stage of PHBV biodegradation but then accelerated subsequent rates of biodegradation. Furthermore, it led to significant increases in total bacterial and fungal biomass and altered the composition of microbial communities with significant increases in the relative abundances of Thauera (gammaproteobacterial) and Mucor circinelloides (fungal) populations. It is proposed, with evidence from biodegradation behavior and microbial analysis, that the presence of DBP likely stimulated a microbial community shift, introduced higher proportions of more readily degradable amorphous regions from the plasticizing effect, and facilitated access to the bulk polymer matrix for microorganisms or at least their associated enzymes. These effects in combination overcame the initial inhibition effect of the DBP and resulted in a net increase in the rate of biodegradation of PHBV.

Comparison of water-energy trajectories of two major regions experiencing water shortage.

Water shortage, increased demand and rising energy costs are major challenges for the water sector worldwide. Here we use a comparative case study to explore the long-term changes in the system-wide water and associated energy use in two different regions that encountered water shortage. In Australia, South East Queensland (SEQ) encountered a drought from 2001 to 2009, while Perth has experienced a decline in rainfall since the 1970s. This novel longitudinal study quantifies and compares the urban water consumption and the energy use of the water supply systems in SEQ and Perth during the period 2002 to 2014. Unlike hypothetical and long-term scenario studies, this comparative study quantifies actual changes in regional water consumption and associated energy, and explores the lessons learned from the two regions. In 2002, Perth had a similar per capita water consumption rate to SEQ and 48% higher per capita energy use in the water supply system. From 2002 to 2014, a strong effort of water conservation can be seen in SEQ during the drought, while Perth has been increasingly relying on seawater desalination. By 2014, even though the drought in SEQ had ended and the drying climate in Perth was continuing, the per capita water consumption in SEQ (266 L/p/d) was still 28% lower than that of Perth (368 L/p/d), while the per capita energy use in Perth (247 kWh/p/yr) had increased to almost five times that of SEQ (53 kWh/p/yr). This comparative study shows that within one decade, major changes in water and associated energy use occurred in regions that were similar historically. The very different "water-energy" trajectories in the two regions arose partly due to the type of water management options implemented, particularly the different emphasis on supply versus demand side management. This study also highlights the significant energy saving benefit of water conservation strategies (i.e. in SEQ, the energy saving was sufficient to offset the total energy use for seawater desalination and water recycling during the period.). The water-energy trajectory diagram provides a new way to illustrate and compare longitudinal water consumption and associated energy use within and between cities.

The contribution of bacteria to algal growth by carbon cycling.

Algal mass production in open systems is often limited by the availability of inorganic carbon substrate. In this paper, we evaluate how bacterial driven carbon cycling mitigates carbon limitation in open algal culture systems. The contribution of bacteria to carbon cycling was determined by quantifying algae growth with and without supplementation of bacteria. It was found that adding heterotrophic bacteria to an open algal culture dramatically enhanced algae productivity. Increases in algal productivity due to supplementation of bacteria of 4.8 and 3.4 times were observed in two batch tests operating at two different pH values over 7 days. A kinetic model is proposed which describes carbon limited algal growth, and how the limitation could be overcome by bacterial activity to re-mineralize photosynthetic end products.

Environmental Benefits and Burdens of Phosphorus Recovery from Municipal Wastewater.

The environmental benefits and burdens of phosphorus recovery in four centralized and two decentralized municipal wastewater systems were compared using life cycle assessment (LCA). In centralized systems, phosphorus recovered as struvite from the solids dewatering liquid resulted in an environmental benefit except for the terrestrial ecotoxicity and freshwater eutrophication impact categories, with power and chemical use offset by operational savings and avoided fertilizer production. Chemical-based phosphorus recovery, however, generally required more resources than were offset by avoided fertilizers, resulting in a net environmental burden. In decentralized systems, phosphorus recovery via urine source separation reduced the global warming and ozone depletion potentials but increased terrestrial ecotoxicity and salinization potentials due to application of untreated urine to land. Overall, mineral depletion and eutrophication are well-documented arguments for phosphorus recovery; however, phosphorus recovery does not necessarily present a net environmental benefit. While avoided fertilizer production does reduce potential impacts, phosphorus recovery does not necessarily offset the resources consumed in the process. LCA results indicate that selection of an appropriate phosphorus recovery method should consider both local conditions and other environmental impacts, including global warming, ozone depletion, toxicity, and salinization, in addition to eutrophication and mineral depletion impacts.

The effect of additives on the biodegradation of polyhydroxyalkanoate (PHA) in marine field trials.

The persistence of conventional fossil fuel-derived plastics in marine ecosystems has raised significant environmental concerns. Biodegradable plastics are being explored as an alternative. This study investigates the biodegradation behaviour in two marine environments of melt-extruded sheets of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) bioplastic as well as blends of PHBV with a non-toxic plasticiser (triethyl citrate, TEC) and composites of PHBV with wood flour. Samples were submerged for up to 35 weeks in two subtropical marine conditions: on the sandy seabed in the sublittoral benthic zone and the sandy seabed of an open air mesocosm with pumped seawater. Rates of biodegradation, lag times and times to 95 % mass loss (T95) were determined through mass loss data and Gompertz modelling. Mechanisms of biodegradation were studied through changes in molecular weight, mechanical properties and surface features. Results reveal a rapid biodegradation rate for all PHBV samples, demonstrating a range of specific biodegradation rates relative to exposed surface area of 0.03 ± 0.01 to 0.09 ± 0.04 mg.d-1.cm-2. This rapid rate of biodegradation meant that the subtle variations in biodegradation mechanisms across different sample thicknesses and additive compositions had little effect on overall lifetimes, with the T95 for most samples being around 250-350 days, regardless of site, highlighting the robust biodegradability of PHBV in seawater. It was only the PHBV-wood flour composite that showed faster biodegradation, and that was only in the exposed ocean site. The mesocosm site was otherwise shown to be a good model for the open ocean, with very comparable biodegradation rates and changes in mechanical properties over time.

Development of a novel electrochemical system for oxygen control (ESOC) to examine dissolved oxygen inhibition on algal activity.

The development of an Electrochemical System for Oxygen Control (ESOC) for examining algal photosynthetic activity as a function of dissolved oxygen (DO) is outlined. The main innovation of the tool is coulombic titration in order to balance the electrochemical reduction of oxygen with the oxygen input to achieve a steady DO set-point. ESOC allows quantification of algal oxygen production whilst simultaneously maintaining a desired DO concentration. The tool was validated abiotically by comparison with a mass transfer approach for quantifying oxygenation. It was then applied to quantify oxygen inhibition of algal activity. Five experiments, using an enriched culture of Scenedesmus sp. as the inoculum, are presented. For each experiment, ESOC was used to quantify algal activity at a series of DO set-points. In all experiments substantial oxygen inhibition was observed at DO >30 mgO2 L-1. Inhibition was shown to fit a Hill inhibition model, with a common Hill coefficient of 0.22±0.07 L mg-1 and common log10 CI50 of 27.2±0.7 mg L-1. This is the first time that the oxygen inhibition kinetic parameters have been quantified under controlled DO conditions.

The confounding effect of nitrite on N2O production by an enriched ammonia-oxidizing culture.

The effect of nitrite (NO2(-)) on the nitrous oxide (N2O) production rate of an enriched ammonia-oxidizing bacteria (AOB) culture was characterized over a concentration range of 0-1000 mg N/L. The AOB culture was enriched in a nitritation system fed with synthetic anaerobic digester liquor. The N2O production rate was highest at NO2(-) concentrations of less than 50 mg N/L. At dissolved oxygen (DO) concentration of 0.55 mg O2/L, further increases in NO2(-) concentration from 50 to 500 mg N/L resulted in a gradual decrease in N2O production rate, which maintained at its lowest level of 0.20 mg N2O-N/h/g VSS in the NO2(-) concentration range of 500-1000 mg N/L. The observed NO2(-)-induced decrease in N2O production was even more apparent at increased DO concentration. At DO concentrations of 1.30 and 2.30 mg O2/L, the lowest N2O production rate (0.25 mg N2O-N/h/g VSS) was attained at a lower NO2(-) concentration of 200-250 mg N/L. These observations suggest that N2O production by the culture is diminished by both high NO2(-) and high DO concentrations. Collectively, the findings show that exceedingly high NO2(-) concentrations in nitritation systems could lead to decreased N2O production. Further studies are required to determine the extent to which the same response to NO2(-) is observed across different AOB cultures.

Hazardous state lifetimes of biodegradable plastics in natural environments.

Plastic pollution is a critical problem that has the potential for long-lasting impact. While all plastics eventually break down to at least some degree, they can remain in different transition states, such as microplastics and nanoplastics, for extended periods of time before reaching complete mineralisation to non-hazardous end products. Each of the transition states represents different types of hazards, so it is critical to understand the factors driving the lifetimes of plastics within these states. To do this, we propose a framework for assessing plastic lifetimes in natural environments based on the flow of material through potentially hazardous states: macroplastic and mesoplastic, microplastic, nanoplastic and soluble products. State changes within this framework are underpinned by three key processes: fragmentation, depolymerisation, and bioassimilation, with the pathways for generation of the different plastic states, and the lifetimes within these states, varying widely for individual materials in different environments due to their dependence on polymer material type, form and properties, and environmental factors. The critical factors driving these processes can therefore appear complex, but molecular weight, crystallinity, oxygen and water diffusivity, and inherent polymer chain reactivity (including to enzymes) are key to our understanding. By analysing currently available data that take factors such as these into consideration, we have generated information on the most likely states in which a range of plastics with different environmental degradation behaviour may exist over time in natural environments. Polyethylene (PE), for example, should be expected to fragment and accumulate in the environment as microplastic and nanoplastic. Interestingly, the state-profile for the biodegradable plastic polylactic acid (PLA) is similar, albeit over shorter timeframes. PLA also likely fragments, but then the relatively slow process of abiotic depolymerisation results in accumulation of microplastic and nanoplastic. By contrast, the state-profile for the biodegradable plastic polyhydroxyalkanoate (PHA) would be expected to be very different. The bulk material is less susceptible to embrittlement and fragmentation as a primary path to biodegradation, since the rapid enzyme catalysed depolymerisation of exposed surfaces proceeds in conjunction with bioassimilation.

Simultaneous colour and DON removal from sewage treatment plant effluent: alum coagulation of melanoidin.

The aim of this study was to detect and characterise melanoidin in sewage treatment plant (STP) effluent, and to study the ability of alum coagulation to remove the colour and dissolved organic nitrogen (DON) associated with melanoidin. The melanoidin is non-biodegradable due to the complex cyclic based structure and thus it directly contributes to effluent nitrogen concentrations from the sewage treatment plant (STP). Lowering of effluent total nitrogen limits and the link between colour and chlorinated disinfection by-products have therefore driven a need to understand the structure, properties and treatability of DON species found in STP effluent. The focus of this paper is the refractory coloured, organic nitrogen compound melanoidin. Wetalla STP effluent has relatively high colour (170mg-PtCoL(-1)) and DON (2.5mgL(-1)) for a biological nutrient removal STP, owing to an industrial supply of melanoidin containing molasses fermentation wastewater. Alum coagulation jar tests were performed on synthetic melanoidin solution, STP effluent containing melanoidin (Wetalla, Toowoomba, Australia) and STP effluent free of melanoidin (Merrimac, Gold Coast, Australia) to examine the treatability of melanoidin and its associated colour and DON content when present in STP effluent. The removal of melanoidin from STP effluent resulted in significant colour and DON reduction. An alum dose of 30mgL(-1) as aluminium was sufficient to reach maximum removal of colour (75%), DON (42%) and dissolved organic carbon (DOC) (30%) present in melanoidin containing STP effluent. Alum was shown to preferentially remove DON with a molecular weight >10kDa over small molecular weight DON. Fluorescence excitation-emission matrix examination of the humic compounds present in the STP effluent indicated that melanoidin type humic compounds were more readily removed by alum coagulation than other humic compounds.

Dissolved methane in rising main sewer systems: field measurements and simple model development for estimating greenhouse gas emissions.

At present, the potential generation of methane in wastewater collection systems is ignored under international greenhouse gas (GHG) accounting protocols, despite recent reports of substantial dissolved methane formation in sewers. This suggests that the current national GHG inventories for wastewater handling systems are likely to be underestimated for some situations. This study presents a new catalogue of field data on methane formation in rising main sewerage systems and proposes an empirically-fitted, theoretical model to predict dissolved methane concentrations, based upon the independent variables of pipeline geometry (i.e. surface area to volume ratio, A/V) and hydraulic retention time (HRT). Systems with longer HRT and/or larger A/V ratios are shown to have higher dissolved methane concentrations. This simple predictive model provides a means for water authorities to estimate the methane emissions from other pressurised sewerage systems of similar characteristics.

Thermophilic production of poly(3-hydroxybutyrate-co-3-hydrovalerate) by a mixed methane-utilizing culture.

The production of polyhydroxyalkanoates (PHAs) from methane is limited to mesophiles and thus suffers from high energy requirements for cooling. To address this issue, the use of thermophilic processes is gaining interest, as this strategy may deliver improved economic feasibility for PHA production. This study reports the first thermophilic PHA-producing culture grown on methane at 55 °C in fill-and-draw batch reactors. Harvested cells were incubated with various combinations of methane, propionic acid and valeric acid to assess their capacity for the synthesis of poly(3-hydroxybutyrate) (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). Only PHB was produced when fed with methane alone. The addition of odd-carbon-number fatty acids resulted in higher PHA content with 3 HV fractions in the range of 15-99 mol%, depending on the types of fatty acids added. Acetic acid addition enhanced the synthesis of 3HB monomer, but not of 3 HV. On increasing the temperature to 58 °C, PHA productivity was not significantly affected.

The rate of biodegradation of PHA bioplastics in the marine environment: A meta-study.

There is a reasonably extensive body of literature recording mass loss of polyhydroxyalkanoates (PHAs) (a class of biodegradable plastics) in the natural marine environment. However, to date, this research has been very disparate. Thus, it remains unclear what the timeframe for the biodegradation of such marine biodegradable plastics actually is. The aim of this work was to determine the rate of biodegradation of PHA in the marine environment and apply this to the lifetime estimation of PHA products. This provides the clarification required as to what 'marine biodegradation of PHA' means in practicality and allows the risks and benefits of using PHA to be transparently discussed. It was determined that the mean rate of biodegradation of PHA in the marine environment is 0.04-0.09 mg·day-1·cm-2 (p = 0.05) and that, for example, a PHA water bottle could be expected to take between 1.5 and 3.5 years to completely biodegrade.

Decreasing activated sludge thermal hydrolysis temperature reduces product colour, without decreasing degradability.

Activated sludges are becoming more difficult to degrade in anaerobic digesters, due to the implementation of stricter nitrogen limits, longer sludge ages, and removal of primary sedimentation units. Thermal hydrolysis is a popular method to enhance degradability of long-age activated sludge, and involves pressure and heat treatment of the process fluid (150-160 degrees C saturated steam). However, as documented in this study, in a full-scale system, the use of thermal hydrolysis produces coloured, recalcitrant compounds that can have downstream impacts (e.g., failure of UV disinfection, and increased effluent nitrogen). The coloured compound formed during thermal hydrolysis was found to be melanoidins. These are coloured recalcitrant compounds produced by polymerisation of low molecular weight intermediates, such as carbohydrates and amino compounds at elevated temperature (Maillard reaction). By decreasing the THP operating temperature from 165 degrees C to 140 degrees C, THP effluent colour decreased from 12,677 mg-PtCo L(-1) to 3837 mg-PtCo L(-1). The change in THP operating temperature from 165 degrees C to 140 degrees C was shown to have no significant impact on anaerobic biodegradability of the sludge. The rate and extent of COD biodegradation remained largely unaffected by the temperature change with an average first order hydrolysis rate of 0.19 d(-1) and conversion extent of 0.43 g-COD(CH4)g-COD(-1).

The degradation of dissolved organic nitrogen associated with melanoidin using a UV/H2O2 AOP.

The aim of this study was to examine the simultaneous degradation of dissolved organic nitrogen (DON) and associated colour from wastewater containing melanoidins by an advanced oxidation process (AOP). UV irradiation of H2O2 was used as the mechanism to create the hydroxyl radical for oxidation. Melanoidins are large nitrogenous organic compounds that are refractory during biological wastewater treatment processes. The simultaneous degradation of DON and colour, present as a result of these compounds, was investigated using an AOP. The oxidation process was much more capable of removing colour (99% degradation), dissolved organic carbon (DOC) (50% degradation) and DON (25% degradation) at the optimal applied dose of hydrogen peroxide for the system (3300 mg l(-1)). This indicated that colour and DON removal were decoupled problems for the purpose of treating melanoidin by an AOP and thus colour removal can not be used as an indication of DON removal Colour was caused by organic molecules with molecular weight greater than 10 kDa. Oxidation caused a partial reduction of the DON (41-15% of the total dissolved nitrogen) and DOC (29-14% of the DOC) associated with the large molecular weight fraction (>10 kDa) and almost complete colour removal (87-3% of the total colour). The degraded DON was mostly accounted for by the formation of ammonia (31% of the nitrogen removed from the large fraction) and small molecular weight compounds (66% of the nitrogen removed from the large fraction). The degraded DOC appeared to be mostly mineralised (to CO2) with only 20% of the degraded compounds appearing as small molecular weight DOC.

Solids characterisation in an anaerobic migrating bed reactor (AMBR) sewage treatment system.

The aim of this work was to characterise the solids in an anaerobic sewage treatment process. Hindered settling velocity, particle size distributions (PSD), influent and effluent COD(P)/SS and discrete settling velocity distributions were all measured. The anaerobic migrating bed reactor (AMBR) solids were mainly flocculent and had a settling rate equivalent to a good settling activated sludge ( approximately SSVI=60 mL/g). The PSD of the anaerobic solids were very different to PSD for activated sludge flocs, with the anaerobic solids having a modal size an order of magnitude smaller than activated sludge, but a range of particle sizes being two orders of magnitude larger. There was a far greater range in size and structure in the anaerobic solids. The anaerobic process solids were primarily feed solids undergoing VSS destruction (hydrolysis). The biological mass was small. The solids seemed to retain their size as the volatiles were degraded and the density decreased ('skeletons' of the influent particulates). The small fraction of slowly settling solids, which have been identified to have a similar modal size but lower density than the mixed solids in the reactor, pose a solids retention time (SRT) control problem when relying on settling alone for solids retention.

Comparative life cycle assessment and financial analysis of mixed culture polyhydroxyalkanoate production.

A life cycle assessment and financial analysis of mixed culture PHA (PHA(MC)) and biogas production was undertaken based on treating an industrial wastewater. Internal rate of return (IRR) and non-renewable CO(2)eq emissions were used to quantify financial viability and environmental impact. PHA(MC) was preferable to biogas production for treating the specified industrial effluent. PHA(MC) was also financially attractive in comparison to pure culture PHA production. Both PHA production processes had similar environmental impacts that were significantly lower than HDPE production. A large potential for optimisation exists for the PHA(MC) process as financial and environmental costs were primarily due to energy use for downstream processing. Under the conditions used in this work PHA(MC) was shown to be a viable biopolymer production process and an effective industrial wastewater treatment technology. This is the first study of its kind and provides valuable insight into the PHA(MC) process.

City-scale analysis of water-related energy identifies more cost-effective solutions.

Energy and greenhouse gas management in urban water systems typically focus on optimising within the direct system boundary of water utilities that covers the centralised water supply and wastewater treatment systems, despite a greater energy influence by the water end use. This work develops a cost curve of water-related energy management options from a city perspective for a hypothetical Australian city. It is compared with that from the water utility perspective. The curves are based on 18 water-related energy management options that have been implemented or evaluated in Australia. In the studied scenario, the cost-effective energy saving potential from a city perspective (292 GWh/year) is far more significant than that from a utility perspective (65 GWh/year). In some cases, for similar capital cost, if regional water planners invested in end use options instead of utility options, a greater energy saving potential at a greater cost-effectiveness could be achieved in urban water systems. For example, upgrading a wastewater treatment plant for biogas recovery at a capital cost of $27.2 million would save 31 GWh/year with a marginal cost saving of $63/MWh, while solar hot water system rebates at a cost of $28.6 million would save 67 GWh/year with a marginal cost saving of $111/MWh. Options related to hot water use such as water-efficient shower heads, water-efficient clothes washers and solar hot water system rebates are among the most cost-effective city-scale opportunities. This study demonstrates the use of cost curves to compare both utility and end use options in a consistent framework. It also illustrates that focusing solely on managing the energy use within the utility would miss substantial non-utility water-related energy saving opportunities. There is a need to broaden the conventional scope of cost curve analysis to include water-related energy and greenhouse gas at the water end use, and to value their management from a city perspective. This would create opportunities where the same capital investment could achieve far greater energy savings and greenhouse gas emissions abatement.

Life-cycle energy impacts for adapting an urban water supply system to droughts.

In recent years, cities in some water stressed regions have explored alternative water sources such as seawater desalination and potable water recycling in spite of concerns over increasing energy consumption. In this study, we evaluate the current and future life-cycle energy impacts of four alternative water supply strategies introduced during a decade-long drought in South East Queensland (SEQ), Australia. These strategies were: seawater desalination, indirect potable water recycling, network integration, and rainwater tanks. Our work highlights the energy burden of alternative water supply strategies which added approximately 24% life-cycle energy use to the existing supply system (with surface water sources) in SEQ even for a current post-drought low utilisation status. Over half of this additional life-cycle energy use was from the centralised alternative supply strategies. Rainwater tanks contributed an estimated 3% to regional water supply, but added over 10% life-cycle energy use to the existing system. In the future scenario analysis, we compare the life-cycle energy use between "Normal", "Dry", "High water demand" and "Design capacity" scenarios. In the "Normal" scenario, a long-term low utilisation of the desalination system and the water recycling system has greatly reduced the energy burden of these centralised strategies to only 13%. In contrast, higher utilisation in the unlikely "Dry" and "Design capacity" scenarios add 86% and 140% to life-cycle energy use of the existing system respectively. In the "High water demand" scenario, a 20% increase in per capita water use over 20 years "consumes" more energy than is used by the four alternative strategies in the "Normal" scenario. This research provides insight for developing more realistic long-term scenarios to evaluate and compare life-cycle energy impacts of drought-adaptation infrastructure and regional decentralised water sources. Scenario building for life-cycle assessments of water supply systems should consider i) climate variability and, therefore, infrastructure utilisation rate, ii) potential under-utilisation for both installed centralised and decentralised sources, and iii) the potential energy penalty for operating infrastructure well below its design capacity (e.g., the operational energy intensity of the desalination system is three times higher at low utilisation rates). This study illustrates that evaluating the life-cycle energy use and intensity of these type of supply sources without considering their realistic long-term operating scenario(s) can potentially distort and overemphasise their energy implications. To other water stressed regions, this work shows that managing long-term water demand is also important, in addition to acknowledging the energy-intensive nature of some alternative water sources.

The Opportunity for High-Performance Biomaterials from Methane.

Polyhydroxyalkanoate (PHA) biopolymers are widely recognised as outstanding candidates to replace conventional petroleum-derived polymers. Their mechanical properties are good and can be tailored through copolymer composition, they are biodegradable, and unlike many alternatives, they do not rely on oil-based feedstocks. Further, they are the only commodity polymer that can be synthesised intracellularly, ensuring stereoregularity and high molecular weight. However, despite offering enormous potential for many years, they are still not making a significant impact. This is broadly because commercial uptake has been limited by variable performance (inconsistent polymer properties) and high production costs of the raw polymer. Additionally, the main type of PHA produced naturally is poly-3-hydroxybutyrate (PHB), which has limited scope due to its brittle nature and low thermal stability, as well as its tendency to embrittle over time. Production cost is strongly impacted by the type of the feedstock used. In this article we consider: the production of PHAs from methanotrophs using methane as a cost-effective substrate; the use of mixed cultures, as opposed to pure strains; and strategies to generate a poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer (PHBV), which has more desirable qualities such as toughness and elasticity.

A laboratory investigation of interactions between denitrifying anaerobic methane oxidation (DAMO) and anammox processes in anoxic environments.

This study investigates interactions between recently identified denitrifying anaerobic methane oxidation (DAMO) and anaerobic ammonium oxidation (anammox) processes in controlled anoxic laboratory reactors. Two reactors were seeded with the same inocula containing DAMO organisms Candidatus Methanoperedens nitroreducens and Candidatus Methylomirabilis oxyfera, and anammox organism Candidatus Kuenenia stuttgartiensis. Both were fed with ammonium and methane, but one was also fed with nitrate and the other with nitrite, providing anoxic environments with different electron acceptors. After steady state reached in several months, the DAMO process became solely/primarily responsible for nitrate reduction while the anammox process became solely responsible for nitrite reduction in both reactors. 16S rRNA gene amplicon sequencing showed that the nitrate-driven DAMO organism M. nitroreducens dominated both the nitrate-fed (~70%) and the nitrite-fed (~26%) reactors, while the nitrite-driven DAMO organism M. oxyfera disappeared in both communities. The elimination of M. oxyfera from both reactors was likely the results of this organism being outcompeted by anammox bacteria for nitrite. K. stuttgartiensis was detected at relatively low levels (1-3%) in both reactors.