Bacteriohopanetetrol-<i>x</i>: constraining its application as a lipid biomarker for marine anammox using the water column oxygen gradient of the Benguela upwelling system

Interpreting lipid biomarkers in the sediment archive requires a good understanding of their application and limitations in modern systems. Recently it was discovered that marine bacteria performing anaerobic ammonium oxidation (anammox), belonging to the genus Ca. Scalindua, uniquely synthesize a stereoisomer of bacteriohopanetetrol (“BHT-x”). The ratio of BHT-x over total bacteriohopanetetrol (BHT, ubiquitously synthesized by diverse bacteria) has been suggested as a proxy for water column anoxia. As BHT has been found in sediments over 50 Myr old, BHT-x has the potential to complement and extend the sedimentary biomarker record of marine anammox, conventionally constructed using ladderane lipids. Yet, little is known about the distribution of BHT-x in relation to the distribution of ladderanes and to the genetic evidence of Ca. Scalindua in modern marine systems. Here, we investigate the distribution of BHT-x and the application of the BHT-x ratio in relation to distributions of ladderane intact polar lipids (IPLs), ladderane fatty acids (FAs) and Ca. Scalindua 16S rRNA genes in suspended particulate matter (SPM) from the water column of the Benguela upwelling system (BUS), sampled across a large oxygen gradient. In BUS SPM, high BHT-x abundances were restricted to the oxygen-deficient zone on the continental shelf (at [O2] < 45 µmol L−1, in all but one case). High BHT-x abundances co-occurred with high abundances of the Ca. Scalindua 16S rRNA gene (relative to the total number of bacterial 16S rRNA genes) and ladderane IPLs. At shelf stations with [O2] > 50 µmol L−1, the BHT-x ratio was < 0.04 (in all but one case). In apparent contradiction, ladderane FAs and low abundances of BHT and BHT-x (resulting in BHT-x ratios > 0.04) were also detected in oxygenated offshore waters ([O2] up to 180 µmol L−1), whereas ladderane IPLs were undetected. The index of ladderane lipids with five cyclobutane rings (NL5) correlates with in situ temperature. NL5-derived temperatures suggested that ladderane FAs in the offshore waters were not synthesized in situ but were transported down-slope from warmer shelf waters. Thus, in sedimentary archives of systems with known lateral organic matter transport, such as the BUS, relative BHT and BHT-x abundances should be carefully considered. In such systems, a higher BHT-x ratio may act as a safer threshold for deoxygenation and/or Ca. Scalindua presence: our results and previous studies indicate that a BHT-x ratio of ≥ 0.2 is a robust threshold for oxygen-depleted waters ([O2] < 50 µmol kg−1). In our data, ratios of ≥ 0.2 coincided with Ca. Scalindua 16S rRNA genes in all samples (n=62), except one. Lastly, when investigating in situ anammox, we highlight the importance of using ladderane IPLs over BHT-x and/or ladderane FAs; these latter compounds are more recalcitrant and may derive from transported fossil anammox bacteria remnants.

Enhanced Nitrogen Removal from Domestic Wastewater by Partial-Denitrification/Anammox in an Anoxic/Oxic Biofilm Reactor

A partial-denitrification coupling with anaerobic ammonium oxidation (anammox) process (PD/A) in a continuous-flow anoxic/oxic (A/O) biofilm reactor was developed to treat carbon-limited domestic wastewater (ammonia (NH4+-N) of 55 mg/L and chemical oxygen demand (COD) of 148 mg/L in average) for about 200 days operation. Satisfactory NH4+-N oxidation efficiency above 95% was achieved with rapid biofilm formation in the aerobic zone. Notably, nitrite (NO2−-N) accumulation was observed in the anoxic zone, mainly due to the insufficient electron donor for complete nitrate (NO3−-N) reduction. The nitrate-to-nitrite transformation ratio (NTR) achieved was as high as 64.4%. After the inoculation of anammox-enriched sludge to anoxic zones, total nitrogen (TN) removal was significantly improved from 37.3% to 78.0%. Anammox bacteria were effectively retained in anoxic biofilm utilizing NO2−-N produced via the PD approach and NH4+-N in domestic wastewater, with the relative abundance of 5.83% for stable operation. Anammox pathway contributed to TN removal by a high level of 38%. Overall, this study provided a promising method for mainstream nitrogen removal with low energy consumption and organic carbon demand.

Enhancing Ammonium Oxidation Fluxes and Nitritation Efficiencies in MABRs: A Modeling Study

The membrane aerated biofilm reactor (MABR) is a novel technology based on gas-supplying membranes that supply dissolved O2 (DO) to biofilms growing on the membrane surface. The counter-diffusion of dissolved...

Addition of anaerobic ammonium oxidation bacteria to lower running cost during the membrane bioreactor process treating sewage

Reducing energy consumption or running cost associated with the membrane bioreactor (MBR) process is a serious challenge that needs to be addressed in treating sewage. The addition of anaerobic ammonium oxidation bacteria (AnAOB) to a running MBR has the potential to lower the aeration rate, thus decreasing the running cost in treating sewage. The results obtained showed that owing to addition of AnAOB, TN and NH4+-N removal rates increased by 9.8% and 1.13%, respectively, while the aeration rate decreased by 50%. Additionally, high throughput sequencing and isotope experiments showed that both AnAOB and heterotrophic denitrification bacteria could survive simultaneously and play an important role in nitrogen removal, with AnAOB having a significantly greater contribution. It can be concluded that the addition of AnAOB reduced the running cost of MBR in treating sewage.

Nitrogen Removal by an Anaerobic Iron-Dependent Ammonium Oxidation (Feammox) Enrichment: Potential for Wastewater Treatment

Nitrogen pollution in water is a growing concern. Anthropogenic activities have increased the amount of nitrogen released into watercourses, which harms human health and the environment, and causes serious problems, such as eutrophication. Feammox is a recently discovered biological pathway associated with the nitrogen cycle that has gained scientific interest. This process couples anaerobic ammonium oxidation with iron reduction. This work presents a study on the Feammox mechanism from the enrichment of an activated sludge obtained from a sewage treatment plant. The enrichment was carried out at neutral pH to study the N2 pathway, that is, the Feammox process with the oxidation of ammonium (NH4+) directly to N2. In addition, different sources of iron were studied: iron chloride (FeCl3); ferrihydrite; and goethite. The characterization of the sludge showed the genes associated with ammonia monooxygenase, nitrate and nitrite reductases processes, along with relevant microbial species. The enrichment, carried out for 42 days and monitored every 14 days, showed that FeCl3 as a source of Fe was more effective for the coupled process of oxidation of NH4+ and the reduction of Fe(III) to Fe(II). At the end of the enrichment period, a removal of 31% and 32.2% of NH4+, and an increase in Fe(II) concentration by 52.4 and 63.9 times regarding the initial value were achieved in aerobic and anaerobic sludge, respectively. This study provides information on the potential of Feammox in the removal of N from wastewater, and the oxidation/reduction yields in the initial enrichment phase.

Recovery profile of anaerobic ammonium oxidation (anammox) bacteria inhibited by ZnO nanoparticles

The potential effects of nanoparticles (NPs) on biological treatment processes have become significant due to their increasing industrial applications. The purpose of this research was to investigate the self-recovery ability of anammox bacteria following to acute ZnO NPs toxicity. In this context, a 2-liter lab-scale anammox reactor was operated for 550 days to enrich the biomass required to the batch exposure tests. Anammox culture was firstly exposed to four different doses of ZnO NPs (50, 75, 100 and 200 mg/L) for 24 h. Then, the ZnO NPs were removed and self-recovery performance of the anammox bacteria was assessed by evaluating the nitrogen removal capacities for 72 h. Besides the nitrogen removal performance, extracellular polymeric substances (EPS) production was also detected to deeply understand the response of the enriched anammox culture against ZnO NPs exposure. The results revealed that, sudden and high load of ZnO NPs (100 and 200 mg/L) resulted in persistent impairment on the nitrogen removal performance of the enriched anammox culture. However, relatively lower doses (50 and 75 mg/L) caused deceleration of the nitrogen removal performance during the recovery period. In addition, EPS content in the reactor decreased along with escalating load of ZnO NPs.