Oxidation Bacteria Research Articles

Cyclic Conversions in the Nitrogen Cycle.

The cyclic nature of specific conversions in the nitrogen cycle imposes strict limitations to the conversions observed in nature and explains for example why anaerobic ammonium oxidation (anammox) bacteria can only use nitrite – and not nitrate – as electron acceptor in catabolism, and why nitrite is required as additional electron donor for inorganic carbon fixation in anabolism. Furthermore, the biochemistry involved in nitrite-dependent anaerobic methane oxidation excludes the feasibility of using nitrate as electron acceptor. Based on the cyclic nature of these nitrogen conversions, we propose two scenarios that may explain the ecological role of recently discovered complete ammonia-oxidizing (comammox) Nitrospira spp., some of which were initially found in a strongly oxygen limited environment: (i) comammox Nitrospira spp. may actually catalyze an anammox-like metabolism using a biochemistry similar to intra-oxic nitrite-dependent methane oxidation, or (ii) scavenge all available oxygen for ammonia activation and use nitrate as terminal electron acceptor. Both scenarios require the presence of the biochemical machinery for ammonia oxidation to nitrate, potentially explaining a specific ecological niche for the occurrence of comammox bacteria in nature.

Development of Strategies for AOB and NOB Competition Supported by Mathematical Modeling in Terms of Successful Deammonification Implementation for Energy-Efficient WWTPs

Novel technologies such as partial nitritation (PN) and partial denitritation (PDN) could be combined with the anammox-based process in order to alleviate energy input. The former combination, also noted as deammonification, has been intensively studied in a frame of lab and full-scale wastewater treatment in order to optimize operational costs and process efficiency. For the deammonification process, key functional microbes include ammonia-oxidizing bacteria (AOB) and anaerobic ammonia oxidation bacteria (AnAOB), which coexisting and interact with heterotrophs and nitrite oxidizing bacteria (NOB). The aim of the presented review was to summarize current knowledge about deammonification process principles, related to microbial interactions responsible for the process maintenance under varying operational conditions. Particular attention was paid to the factors influencing the targeted selection of AOB/AnAOB over the NOB and application of the mathematical modeling as a powerful tool enabling accelerated process optimization and characterization. Another reviewed aspect was the potential energetic and resources savings connected with deammonification application in relation to the technologies based on the conventional nitrification/denitrification processes.

Markers for the Comparison of the Performances of Anoxic Biotrickling Filters in Biogas Desulphurisation: A Critical Review

The agriculture and livestock industry generate waste used in anaerobic digestion to produce biogas containing methane (CH4), useful in the generation of electricity and heat. However, although biogas is mainly composed of CH4 (~65%) and CO2 (~34%), among the 1% of other compounds present is hydrogen sulphide (H2S) which deteriorates engines and power generation fuel cells that use biogas, generating a foul smell and contaminating the environment. As a solution to this, anoxic biofiltration, specifically with biotrickling filters (BTFs), stands out in terms of the elimination of H2S as it is cost-effective, efficient, and more environmentally friendly than chemical solutions. Research on the topic is uneven in terms of presenting performance markers, underestimating many microbiological indicators. Research from the last decade was analyzed (2010–2020), demonstrating that only 56% of the reviewed publications did not report microbiological analysis related to sulphur oxidising bacteria (SOB), the most important microbial group in desulphurisation BTFs. This exposes fundamental deficiencies within this type of research and difficulties in comparing performance between research works. In this review, traditional and microbiological performance markers of anoxic biofiltration to remove H2S are described. Additionally, an analysis to assess the efficiency of anoxic BTFs for biogas desulphurisation is proposed in order to have a complete and uniform assessment for research in this field.

Production of Endotoxin-Free Microbial Biomass for Food Applications by Gas Fermentation of Gram-Positive H2-Oxidizing Bacteria

Production of Endotoxin-Free Microbial Biomass for Food Applications by Gas Fermentation of Gram-Positive H₂-Oxidizing Bacteria

Community Composition and Spatial Distribution of N-Removing Microorganisms Optimized by Fe-Modified Biochar in a Constructed Wetland.

Microbial nitrogen (N) removal capability can be significantly enhanced in a horizontal subsurface flow constructed wetland (HSCW) amended by Fe-modified biochar (FeB). To further explore the microbiological mechanism of FeB enhancing N removal, nirS- and nirK-denitrifier community diversities, as well as spatial distributions of denitrifiers and anaerobic ammonium oxidation (anammox) bacteria, were investigated in HSCWs (C-HSCW: without biochar and FeB; B-HSCW: amended by biochar; FeB-HSCW: amended by FeB) treating tailwater from a wastewater treatment plant, with C-HSCW without biochar and FeB and B-HSCW amended by biochar as control. The community structures of nirS- and nirK-denitrifiers in FeB-HSCW were significantly optimized for improved N removal compared with the two other HSCWs, although no significant differences in their richness and diversity were detected among the HSCWs. The spatial distributions of the relative abundance of genes involved in denitrification and anammox were more heterogeneous and complex in FeB-HSCW than those in other HSCWs. More and larger high-value patches were observed in FeB-HSCW. These revealed that FeB provides more appropriate habitats for N-removing microorganisms, which can prompt the bacteria to use the habitats more differentially, without competitive exclusion. Overall, the Fe-modified biochar enhancement of the microbial N-removal capability of HSCWs was a result of optimized microbial community structures, higher functional gene abundance, and improved spatial distribution of N-removing microorganisms.

Culturing sludge fermentation liquid-driven partial denitrification in two-stage Anammox process to realize advanced nitrogen removal from mature landfill leachate

The two-stage partial nitrification (PN)-Anammox process, during long term treatment of high-ammonia nitrogen leachate, faces challenges such as the adaptation of nitrite oxidation bacteria (NOB) and failure of real-time control of pH. Resultant instabilities including NH4+-N and NO3--N accumulation were overcome by culturing sludge fermentation liquid (SFL)-driven partial denitrification (PD) in situ in the Anammox process. Biodegradation of slowly biodegradable organics (SBO) in SFL created organics restriction condition, which limited the activity of denitrification bacteria and achieved its balance with Anammox bacteria. Produced NO3--N is reduced to NO2--N through PD, which further improved the removal of NH4+-N through Anammox. NO2--N was utilized timely by Anammox bacteria, which avoid further reduction of NO2--N to N2, and result in a high nitrate to nitrite transformation ratio (NTR) of 93.3%. Satisfactory nitrogen removal efficiency (NRE) and nitrogen removal rate (NRR) of 99.6% and 822.0 ± 9.0 g N/(m3∙d) were obtained, respectively. Key genera related to degradation of SBO, PD and Anammox were enriched. The value of narG/(nirK+nirS) increased from 0.05 on day 1–0.15 on day 250. Combining SFL-driven PD with two-stage Anammox process provided a novel insight for applying this process to realize advanced nitrogen removal in practical engineering.

Integration of Phosphate Solubilising Bacteria, Sulphur Oxidizing Bacteria with NPK on Maize (Zea mays)

Deficiency of phosphorus can reduce the yield up to 15-20% and its availability to crop plants is the main issue, particularly in calcareous soils of Pakistan. Micro-organisms, phosphorus solubilizing bacteria (PSB) and sulphur oxidizing bacteria (SOB) have the ability to make P accessible for optimum plant growth, under conditions of nutrients disparity. Therefore, six treatments i.e. control, Yz dose of NPK, full recommended dose of NPK, Yz NPK+ SOB, Yz NPK + PSB and Yz NPK + SOB+ PSB were applied in a field experiment to investigate the integrated role of SOB and PSB with NPK fertilizers for enhancing the maize production. The treatment was concluded to have major impact on agro-morphological traits, seed quality and growth parameters of maize. Results depicted that the use of Yz NPK + SOB + PSB gave maximum germination count/plot (151.33), plant height (189.03 cm), number of ears/plant (1.60), grains/ear (472.33), 1000-grain weight (305.67 g), grain yield (5350.50 Kg/ha), harvest index (31.23%), leaf area (379.77 cm2), total dry matter accumulated (181.43 g/plant), crop growth rate (30.60 g/day), net assimilation rate (9.31 g/day) and protein contents (8.49%).
 

Potential microbial functions and quorum sensing systems in partial nitritation and anammox processes

Diverse microbial communities coexist in the partial nitritation-anaerobic ammonium oxidation (PNA) process, in which nitrogen metabolism and information exchange are two important microbial interactions. In the PNA process, the existence of diverse microorganisms including nitrifiers, anammox bacteria, and heterotrophs makes it challenging to achieve a balanced relationship between anaerobic ammonium oxidation bacteria and ammonia oxidizing bacteria. In this study, potential microbial functions in nitrogen conversion and acyl-homoserine lactones (AHLs)-based quorum sensing (QS) in PNA processes were examined. Candidatus_Kuenenia and Nitrosomonas were the key functional bacteria responsible for PNA, while Nitrospira was detected as the dominant nitrite oxidizing bacteria (NOB). Heterotrophs containing nxr might play a similar function to NOB. The AHLs-QS system was an important microbial communication pathway in PNA systems. N-octanoyl-L-homoserine lactone, N-decanoyl homoserine lactone, and N-dodecanoyl homoserine lactone were the main AHLs, which might be synthesized by nitrogen converting microorganisms and heterotrophs. However, only heterotrophs had the potential to sense and degrade AHLs, such as Saccharophagus (sensing) and Leptospira (degradation). These results provide comprehensive information about the possible microbial functions and interactions in the PNA system and clues for system optimization from a microbial perspective. PRACTITIONER POINTS: ●Potential functions of anammox bacteria, nitrifiers, and heterotrophs were revealed. ●Diverse nitrogen conversion and AHLs-quorum sensing related genes were detected. ●Anammox bacteria and AOB played important roles in the AHLs synthesis process. ●Heterotrophs could sense and degrade AHLs during information exchange.

The screening of early warning indicators and microbial community of chicken manure thermophilic digestion at high organic loading rate

Anaerobic digestion of chicken manure (CM) with high nitrogen content is easily suffered inhibition of ammonia, especially under thermophilic and high organic loading rate (OLR) condition. To screen the early warning indicators for instability, thermophilic digestion of CM was conducted in a bench-scale reactor (working volume 55 L) under OLR from 1.0 to 6.0 g VS/(L·d). Microbial community analysis was used to reveal the mechanism of instability at high OLR. The system collapsed at an OLR of 6.0 g VS/(L·d). The initial ammonia inhibition evoked serious acidification, subsequently, the dual suppression of free volatile fatty acids and free ammonia caused the final collapse. The most likely mechanism of iso-butyrate degradation was propionate pathway. The ratio of n-butyrate to iso-butyrate was screened as an early warning indicator with the threshold of above 0.45. The high OLR of 6.0 g VS/(L·d) broke the balance of syntrophic oxidation bacteria (norank_o__MBA03, Gelria, and norank_o__D8A-2), hydrogenotrophic methanogens (Methanothermobacter) and fermentative and acid-producing bacteria (Ruminiclostridium_1, Bacteroides, Defluviitalea, norank_o__Clostridia, and norank_o__M55-D21). The non-functional acid tolerant bacteria (Defluviitoga, norank_f__Family_XI, Corynebacterrium_1, Caldicoprobacter, Lactobacillus, and Erysipelothrix) became the dominant after the system collapsed.

Rapid formation of granules coupling n-DAMO and anammox microorganisms to remove nitrogen

Granular sludge exhibits unique features, including rapid settling velocity, high loading rate and relative insensitivity against inhibitors, thus being a favorable platform for the cultivation of slow-growing and vulnerable microorganisms, such as anaerobic ammonium oxidation (anammox) bacteria and nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) microorganisms. While anammox granules have been widely applied, little is known about how to speed up the granulation process of n-DAMO microorganisms, which grow even slower than anammox bacteria. In this study, we used mature anammox granules as biotic carriers to embed n-DAMO microorganisms, which obtained combined anammox + n-DAMO granules within 6 months. The results of whole-granule 16S rRNA gene amplicon sequencing showed the coexistence of anammox bacteria, n-DAMO bacteria and n-DAMO archaea. The microbial stratification along granule radius was further elucidated by cryosection-16S rRNA gene amplicon sequencing, showing the dominance of n-DAMO archaea and anammox bacteria at inner and outer layers, respectively. Moreover, the images of cryosection-fluorescence in situ hybridization (FISH) verified this stratification and also indicated a shift in microbial stratification. Specifically, n-DAMO bacteria and n-DAMO archaea attached to the anammox granule surface initially, which moved to the inner layer after 4-months operation. On the basis of combined anammox + n-DAMO granules, a practically useful nitrogen removal rate (1.0 kg N/m3/d) was obtained from sidestream wastewater, which provides new avenue to remove nitrogen from wastewater using methane as carbon source.

One-stage partial nitrification and anammox process in a sequencing batch biofilm reactor: Start-up, nitrogen removal performance and bacterial community dynamics in response to temperature

A one-stage partial nitrification and anammox (PN/A) process was started up and operated under varying temperatures in a lab-scale sequencing batch biofilm reactor. The start‑up phase took 110 days with an intermittent aeration strategy, and the removal efficiencies of ammonia‑nitrogen and total nitrogen were found to be 92.22% and 76.07%, respectively. The total nitrogen removal efficiency (NRE) increased by 9.49% when temperature decreased from 30 °C to 25 °C, but declined by 83.84% from 25 °C to 20 °C. The PN process was inhibited and subsequently limited the nitrogen removal performance at 20 °C. When temperature returned to 28 °C, the NRE recovered to 67.27%, but it was still lower than the value before the decrease in temperature (79.40%). Microbial community analysis showed that the predominant ammonia oxidation bacteria and anammox bacteria were Nitrosomonas and Candidatus Kuenenia, respectively. Nitrosomonas grew, while the relative abundance of Candidatus Kuenenia increased as temperature decreased and vice versa.

Comparison of Ammonium-oxidizing Bacterial Community Changes in Sludges from a Sewage and a Marine Fish Market Wastewater Treatment Plant During Enrichment Cultivation Under High Saline Conditions

Objectives:It is important to enrich and cultivate ammonia oxidation bacteria (AOB) in order to successfully treat nitrogen in high saline wastewater using a deammonification process. Two different inocula, a sewage sludge and a fish-market wastewater sludge, were cultivated to enrich AOB and compared the changes of microbial community.<br/>Methods:A sequential batch reactor (SBR) inoculated with the sewage sludge (PN1) enriched AOB under high-strength ammonium condition (500-3,000 mg NH4<sup>+</sup>-N/L) and then the salt concentration in the medium was gradually increased up to 20 g-NaCl/L. The other SBR seeded with the fish market wastewater sludge (PN2) was operated to enrich AOB directly under 20 g NaCl/L without any acclimation step.<br/>Results and Discussion:Both PN1 and PN2 successfully showed more than 60% of the nitrite accumulation efficiency at a high saline concentration of 20 g NaCl/L. At the level of the phylum, Proteobacteria containing nitrifying microorganisms became dominant in both PN1 and PN2. However, the most dominant bacterial species in PN1 and PN2 were Nitrosomonas eutropha (60.7%) and N. halophila (20.2%), respectively.<br/>Conclusions:Although different results of the most abundant AOB were shown in both sewage sludge and fish-market wastewater sludge conditions, nitritation was successfully developed even with wastewater containing high salinity. Therefore, in this study, both sludges can be applied for inoculation to the PN process for efficiently treating wastewater with high concentration of ammonium and saline.

Effects of biotin on promoting anammox bacterial activity

Anaerobic ammonium oxidation (anammox) bacteria significantly improve the efficiency and reduce cost of nitrogen removal in wastewater treatment plants. However, their slow growth and vulnerable activity limit the application of anammox technology. In this paper, the enhancement of biotin on the nitrogen removal activity of anammox bacteria in short-term batch experiments was studied. We found that biotin played a significant role in promoting anammox activity within a biotin concentration range of 0.1–1.5 mg/L. At a biotin concentration of 1.0 mg/L, the total nitrogen removal rate (NRR) increased by 112%, extracellular polymeric substance (EPS) secretion and heme production significantly improved, and anammox bacterial biomass increased to maximum levels. Moreover, the predominant genus of anammox bacteria was Candidatus Brocadia.

Effect of biomass immobilization and reduced graphene oxide on the microbial community changes and nitrogen removal at low temperatures

The slow growth rate and high optimal temperatures for the anaerobic ammonium oxidation (anammox) bacteria are significant limitations of the anammox processes application in the treatment of mainstream of wastewater entering wastewater treatment plant (WWTP). In this study, we investigate the nitrogen removal and microbial community changes in sodium alginate (SA) and sodium alginate–reduced graphene oxide (SA-RGO) carriers, depending on the process temperature, with a particular emphasis on the temperature close to the mainstream of wastewater entering the WWTP. The RGO addition to the SA matrix causes suppression of the beads swelling, which intern modifies the mechanical properties of the gel beads. The effect of the temperature drop on the nitrogen removal rate was reduced for biomass entrapped in SA and SA-RGO gel beads in comparison to non-immobilized biomass, this suggests a ‘‘protective” effect caused by immobilization. However, analyses performed using next-generation sequencing (NGS) and qPCR revealed that the microbial community composition and relative gene abundance changed significantly, after the implementation of the new process conditions. The microbial community inside the gel beads was completely remodelled, in comparison with inoculum, and denitrification contributed to the nitrogen transformation inside the beads.

Pilot-scale enrichment of anammox biofilm using secondary effluent as source water.

Enough biomass of anaerobic ammonium oxidation (anammox) bacteria is essential for maintaining a stable partial nitrification/anammox (PN/A) wastewater treatment system. Present enrichment procedures are mainly labor-intensive and inconvenient for up-scaling. A simplified procedure was developed for enrichment of anammox biofilm by using secondary effluent as source water with no supplement of mineral medium and unstrict control of influent dissolved oxygen (DO). Anammox biofilm was successfully enriched in two pilot-scale reactors (XQ-cul and BT-cul) within 250 and 120 days, respectively. The specific anammox activity increased rapidly during the last 2 months in both reactors and achieved 2.54 g N2-N/(m2·d) in XQ-cul and 1.61 g N2-N/(m2·d) in BT-cul. Similar microbial diversity and community structure were obtained in the two reactors despite different secondary effluent being applied from two wastewater treatment plants. Anaerobic ammonium oxidizing bacteria genera abundance reached up to 37.4% and 43.1% in XQ-cul and BT-cul biofilm, respectively. Candidatus Brocadia and Ca. Kuenenia dominated the enriched biofilm. A negligible adverse effect of residual organics and influent DO was observed by using secondary effluent as source water. This anammox biofilm enrichment procedure could facilitate the inoculation and/or bio-augmentation of large-scale mainstream PN/A reactors.

Key factors governing the performance and microbial community of one-stage partial nitritation and anammox system with bio-carriers and airlift circulation

A one-stage airlift internal circulation biofilm reactor was continuously operated for 668 days to treat 50 mg/L of ammonia wastewater to pursue the long-term stability of partial nitritation and anammox (PNA) process. The operational performance and microbial community structure of the biofilm and the flocs were investigated. A nitrogen removal efficiency (NRE) of 70% was obtained successfully at a dissolved oxygen (DO) of 0.05–0.15 mg/L by regulating aeration rate. The microbial analysis indicated Candidatus Brocadia (29.5%) and Nitrosomonas (6.8%) were dominant in both biofilms and flocs. It was found that DO control and aeration rate were the key factors in performance stability, and a stable performance could be recovered and maintained under oxygen-limiting conditions. Further, the achievement of activated ammonia oxidation bacteria (AOB), dominated anammox bacteria (AMX), suppressed NOB, and controlled heterotrophic bacteria (HB) in the biofilms played a major role in the long-term stable operation.

Rapid enrichment of anammox bacteria linked to floc aggregates in a single-stage partial nitritation-anammox process: Providing the initial carrier and anaerobic microenvironment

Rapid enrichment of anaerobic ammonia oxidation bacteria (AnAOB) is highly associated with the granulation process; however, the interactive mechanism remains unclear, especially for the initial granulation process. A single-stage partial nitritation-anammox (PN/A) bioreactor combined with granular/floc sludge was operated for 400 days. During the experimental period, the nitrogen removal rate increased from 0.60 to 1.21 kg N m−3d−1, and the nitrogen removal capability improved primarily during a transition period (days 200–250), which was accompanied by a particle size increase and AnAOB proliferation (4.9 ± 1.7 days). Moreover, as observed by the biomass physio-morphology, the size distribution, and the microbial community shift, small flocs (< 200 μm) aggregated due to the addition of excess sodium acetate. The emerging floc aggregates represented an early form of granules, providing the initial biological carrier and necessary anaerobic microenvironment for the growth of attached AnAOB, resulting in a high AnAOB growth-rate. These results are the first direct evidence that floc aggregates are essential to AnAOB enrichment, and that they can be affected by operational conditions. This study provides an in-depth understanding of the link between floc aggregations and AnAOB enrichment and broadens the feasibility of optimizing PN/A applications.

Distribution and Community Composition of Anammox Bacteria in Shallow Groundwater of the Kathmandu Valley, Nepal.

The abundance and diversity of anaerobic ammonium oxidation (anammox) bacteria were assessed in 152 groundwater samples in the Kathmandu Valley, Nepal. Anammox bacterial 16S rRNA genes were detected in 54% (37/68) of samples collected in the dry season at 1.6×105–8.8×106 copies L–1, and in 60% (50/84) of samples collected in the wet season at 4.3×104–1.2×107 copies L–1. The 16S rRNA genes of “Candidatus Brocadia”, “Candidatus Anammoxoglobus”, and five new deduced anammox bacterial phylotypes were detected in the shallow groundwater samples. Diverse anammox bacteria were broadly distributed in the shallow groundwater aquifer of the Kathmandu Valley.

The shelf life of buffalo meat marinated with pomegranate (Punica granatum) peel extract.

Objective:The purpose of this study was to investigate how pomegranate peel extract (PPE) can prevent lipid oxidation, peroxide value, and pathogenic bacteria growth in buffalo meat.Materials and Methods:PPE and buffalo meat were employed in this investigation. The buffalo meat marinated with PPE was evaluated by refrigerating it at a temperature of 5°C ± 1°C on days 0, 4, 8, 12, and 16. PPE was added to buffalo meat at a rate of 0% as a control (K0), 0.50% (K1), 1.00% (K2), 1.50% (K3), and 2.00% (K4).Results:The addition of PPE lowered the total plate count, peroxide value, lipid, and pH between treatments and storage period (p < 0.05). PPE’s high concentration of polyphenols, flavonoids, antioxidants, and antibacterial substances may decrease lipid oxidation, peroxide production, and bacterial growth rate.Conclusions:Marinating buffalo meat in PPE may help maintain the meat’s freshness while being stored at a refrigerator temperature (5°C ± 1°C).

Digestate Application on Two Different Soils: Agricultural Benefit and Risk

In order to valorize the use of digestate, the purposes of this study was to discriminate the fertilizing potential of solid and liquid fractions of digestate using two soils that differed for chemical characteristics, to expand the digestate use reducing its environmental impact. The two fractions did not contain toxic compounds and differed in chemical compositions. The two soils responded differently to the addition of the two-digestate fractions and the benefit depended mainly on soil characteristics rather than on quantity and quality of the organic material applied. In the soil with neutral pH, the highest intrinsic amount of organic matter, microbial biomass (MBC), fungi, bacteria and cation exchange capacity were observed; all these properties increased the most over time, in presence of both solid and liquid fractions. Differently, in the soil with alkaline pH and minor amount of intrinsic organic matter, MBC, fungi and bacteria, only few properties such as oxidative soil activity, bacteria colonies, and organic matter amount were improved by the addition of digestate fractions. The use of both fractions showed more agricultural advantages in respect to the relative risks, and the solid fraction was the most effective. Even if the effects of digestate on soil ecosystem can differ in extent, we can expect economic benefit deriving from the reduction of the costs for its disposal, agricultural benefit for their high supply of nutrients to the soil and environmental advantages for the decrement in the use of manufactured fertilizers.