Fabrication of a novel polyurethane foam-alginate-zeolite hydrogel and subsequent KSND bacteria encapsulation: evidence of accelerated biofilm colonization and enhanced nitrogen removal efficiency.

Biofilms are used widely to remove nitrogen from wastewater; however, most biofilm carriers (i.e. polyurethane foam, PUF) are hydrophobic organic materials with millimetre-scale apertures, ineffective attachment, and unstable colonization of microorganisms. To address these limitations, hydrophilic sodium alginate (SA) mixed with zeolite powder (Zeo) was cross-linked in PUF to form a micro-scale hydrogel (PAS) with a well-organized and reticular cellular structure. Scanning electron microscopy revealed that immobilized cells were entrapped in the interior of hydrogel filaments and rapidly formed a stable biofilm on the surface. The biofilm generated was 10.3-fold greater than the film developed on PUF. Kinetics and isotherm studies revealed that the as-developed carrier, because of the presence of Zeo, effectively improved the adsorption of NH4+-N by 53%. The PAS carrier achieved total nitrogen removal in excess of 86% for low carbon-to-nitrogen ratio wastewater treated for 30 d, indicating that this novel modification-encapsulation technology has potential for wastewater treatment.

Enhanced nitrogen removal from low-temperature wastewater by an iterative screening of cold-tolerant denitrifying bacteria.

The biological process to remove nitrogen in winter effluent is often seriously compromised due to the effect of low temperatures (< 13 °C) on the metabolic activity of microorganisms. In this study, a novel heterotrophic nitrifying-aerobic denitrifying bacterium with cold tolerance was isolated by iterative domestication and named Moraxella sp. LT-01. The LT-01 maintained almost 60% of its maximal growth activity at 10 °C. Under initial concentrations of 100 mg/L, the removal efficiencies of ammonium, nitrate, nitrite by LT-01 were 70.3%, 65.4%, 61.7% respectively for 72 h incubation at 10 °C. Nitrogen balance analysis showed that about 46% of TN was released as gases and 16% of TN was assimilated for cell growth. The biomarker genes involved in nitrification and denitrification pathways were identified by gene-specific PCR and revealed that the LT-01 has nitrite reductase (NirS) but not hydroxylamine reductase (HAO), which implies the involvement of other genes in the process. The study indicates that LT-01 has the potential for use in low-temperature regions for efficient sewage treatment.

A high-yielding strain of indole-3-acetic acid isolated from food waste compost: metabolic pathways, optimization of fermentation conditions, and application.

Food waste is a potential resource to prepare microbial fertilizer. However, functional microorganisms derived from the food waste compost (FWC) are relatively lacking. We have isolated, identified, characterized and optimized a high-yielding indole-3-acetic acid (IAA) strain from FWC and further evaluated its growth promoting effect on plants. A IAA high-yielding strain, Providencia sp.Y, with an initial IAA yield of 139.98 mg L-1, was obtained through high-throughput screening, and identified by 16S rRNA gene sequence. The novel strain Y may simultaneously involve the following three pathways from L-tryptophan to IAA, which were identified using liquid chromatography-tandem mass spectrometry: (1) L-tryptophan-indole-3-ethanol-indole-3-acetaldehyde-indole-3-acetic acid; (2) L-tryptophan-1-hydroxy-indole-3-ethanol-indole-3-acetic acid; (3) L-tryptophan-indole-3-acetamide-indole-3-acetic acid. The most suitable comprehensive conditions for IAA production, which were optimized by single factor experiment, were: culture time 12 h, inoculation amount 2% (v/v), NaCl concentration 4% (w/v), culture temperature 25℃, initial pH = 5, and L-tryptophan concentration 3.0 g L-1. The yield of IAA after optimization was increased by 590.48%, from 139.98 mg L-1 (before optimization) to 966.54 mg L-1. Diluted 200-fold microbial suspension could significantly improve the growth of pakchoi seedlings. The seedling plant height, root length, leaf width, leaf length, and fresh weight with microbial suspension increased by 17.39%, 107.35%, 77.98%, 37.75%, and 215.38%, respectively, compared with those without microbial suspension. The increase was greater than that of commercial bacterial agents. In conclusion, this isolated strain can be used as an economical microbial inoculant and provides a new germplasm resource for developing microbial fertilizers.

New insight into the metabolic mechanism of a novel lipid-utilizing and denitrifying bacterium capable of simultaneous removal of nitrogen and grease through transcriptome analysis.

Introduction: Issues related to fat, oil, and grease from kitchen waste (KFOG) in lipid-containing wastewater are intensifying globally. We reported a novel denitrifying bacterium Pseudomonas CYCN-C with lipid-utilizing activity and high nitrogen-removal efficiency. The aim of the present study was aim to explore the metabolic mechanism of the simultaneous lipid-utilizing and denitrifying bacterium CYCN-C at transcriptome level. Methods: We comparatively investigated the cell-growth and nitrogen-removal performances of newly reported Pseudomonas glycinae CYCN-C under defined cultivation conditions. Transcriptome analysis was further used to investigate all pathway genes involved in nitrogen metabolism, lipid degradation and utilization, and cell growth at mRNA levels. Results: CYCN-C could directly use fat, oil, and grease from kitchen waste (KFOG) as carbon source with TN removal efficiency of 73.5%, significantly higher than that (60.9%) with sodium acetate. The change levels of genes under defined KFOG and sodium acetate were analyzed by transcriptome sequencing. Results showed that genes cyo, CsrA, PHAs, and FumC involved in carbon metabolism under KFOG were significantly upregulated by 6.9, 0.7, 26.0, and 19.0-folds, respectively. The genes lipA, lipB, glpD, and glpK of lipid metabolic pathway were upregulated by 0.6, 0.4, 21.5, and 1.3-folds, respectively. KFOG also improved the denitrification efficiency by inducing the expression of the genes nar, nirB, nirD, and norR of denitrification pathways. Conclusion: In summary, this work firstly provides valuable insights into the genes expression of lipid-utilizing and denitrifying bacterium, and provides a new approach for sewage treatment with reuse of KFOG wastes.

Reuse of composted food waste from rural China as vermicomposting substrate: effects on earthworms, associated microorganisms, and economic benefits.

ABSTRACTAerobic composting of food waste (FW) from rural China using a composting device results in a substantial financial burden on the government. This study aimed to assess the feasibility of mitigating this cost using vermicomposting of composted FW. The specific aims were to elucidate the effects of composted FW on earthworm growth and reproduction, reveal the changes in the physical and chemical properties of earthworm casts during vermicomposting, identify the microbial community structure associated with vermicomposting, and perform a financial analysis based on the yield of earthworms and earthworm casts. Mixing composted FW and mature cow dung in an equal ratio achieved the highest earthworm reproduction rate, where 100 adult earthworms produced 567 juvenile earthworms and 252 cocoons in 40 d. Earthworms reduce salt content of vermicomposting substrates by assimilating Na+ and promoting humification by transforming humin into humic and fulvic acid, thus producing earthworm casts with a high generation index > 80%. When composted FW was added to a vermicomposting substrate, a distinctive microbial community structure with alkaliphilic, halophilic, and lignocellulolytic microorganisms dominated the microflora. The dominant bacterial species was Saccharopolyspora rectivirgula, and the dominant fungal species changed from Kernia nitida to Coprinopsis scobicola. Furthermore, microbial genes for refractory organic matter and fat degradation were observed in Vibrio cholerae, Kernia nitida, and Coprinopsis scobicola. Financial analysis showed that vermicomposting has the potential to reduce the cost associated with FW disposal from $ 57 to $ 18/t.

[Aeromonas immobilized on chitosan for treating high-oil wastewater from kitchens].

To effectively solve the serious impact of high oil in the kitchen wastewater on the downstream treatment process, an excellent oil-degrading strain Aeromonas allosaccarophila CY-01 was immobilized to prepare Chitosan-Aeromonas pellets (CH-CY01) by using chitosan as a carrier. Oil degradation condition and efficiency of CH-CY01 pellets were assessed. The growth of immobilized CH-CY01 was almost unaffected, and the maximum degradation rate of soybean oil was 89.7%. Especially at 0.5% NaCl concentration, oil degradation efficiency of CH-CY01 was increased by 20% compared with free cells. In the presence of a surfactant (sodium dodecylbenzene sulfonate) at 1 mg/L, the degradation efficiency of oil by CH-CY01 was increased by 40%. Moreover, using the high-oil catering wastewater as the substrate, more than 80% of the solid oil was degraded with 1% (V/V) CH-CY01 pellets treatment for 7 days, significantly higher than that of free cells. In summary, immobilized CH-CY01 significantly improved the efficiency of oil degradation.

Enhanced stability and nitrogen removal efficiency of Klebsiella sp. entrapped in chitosan beads applied in the domestic sewage system.

Although numerous denitrifying bacteria have been isolated and characterized, their capacity is seriously compromised by traditional inoculant addition and environmental stress in open bioreactors for wastewater treatment. In this study, a biocompatible material, chitosan, was used as a carrier to immobilize a simultaneously heterotrophic nitrifying-aerobic denitrifying bacterium Klebsiella sp., KSND, for continuous nitrogen removal from domestic wastewater in an open purification tank. The results showed that immobilization had no significant effect on cell viability and was beneficial for the reproduction and adhesion of cells. The entrapped KSND exhibited a slightly higher nitrogen removal efficiency of 90.09% than that of free KSND (87.69%). Subsequently, repeated batch cultivation experiments and analysis of the effects of organic contaminants and metal ions were performed using artificial wastewater and domestic wastewater. The findings revealed that the immobilized KSND beads presented desirable biophysical properties with good mechanical stability, cell viability, and enrichment, remarkable stability in organic contaminants and metal ions, and high efficiency nitrogen removal capacity. In conclusion, the developed immobilized denitrifying bacteria system has great potential for continuous wastewater treatment in open bioreactors.

New insight into the nitrogen metabolism of simultaneous heterotrophic nitrification-aerobic denitrification bacterium in mRNA expression.

Here, the draft genome of simultaneous nitrification-denitrification strain (SND) Klebsiella sp. KSND revealed possible existence of genes involved in N-assimilation and -dissimilation pathways. The change levels of genes under defined N-sources were analyzed by Quantitative Real-Time PCR. It suggested that NH4+-assimilation via NADP-glutamate dehydrogenase pathway would occur preferentially. NirBD genes were tightly regulated in a lower level, so that nitrite was rapidly consumed for detoxication by denitrification. Three types of nitrate reductase homologues are surprisingly present in KSND, whereas the dominant nitrate reduction for assimilation and denitrification processes mediates by NapA-type nitrate reductase. Nitric oxide reductase homologues FlRd and FlRd-red provide an adequate capacity for NO detoxification. The recombinant hydroxylamine reductase showed high activity in hydroxylamine to generate ammonium, which might contribute to detoxification mechanism in nitrogen cycling. Overall, this study firstly provides valuable insights into the genes expression and enzyme action, which helps understanding the mechanism of SND processes.

Efficient nitrogen removal by simultaneous heterotrophic nitrifying-aerobic denitrifying bacterium in a purification tank bioreactor amended with two-stage dissolved oxygen control.

Nitrogen removal performance of a simultaneous heterotrophic nitrifying-aerobic denitrifying (SND) bacterium (KSND) in a purification tank bioreactor (PTBR) amended with two-stage dissolved oxygen (DO) control was investigated. NH4+-N and total nitrogen (TN) removal efficiencies under aerobic conditions for domestic wastewater treatment were 97.12% and 52.64%, respectively. Under serial aerobic (DO > 4.0 mg/L) and anaerobic (DO < 0.5 mg/L) phases, average TN removal efficiency from effluent was 95.45%, without nitrate and nitrite accumulation. DO control assay demonstrated that anaerobic condition adversely affected nitrification (46.13%), but was conducive to denitrification (93.52%). Transcriptional analysis revealed 2.72-fold increase in hydroxylamine reductase expression under aerobic condition as compared to anaerobic condition. Nitrate reductase and nitric oxide reductase homologs had the additional activity of supporting anaerobic or aerobic denitrification in SND bacteria. Under two-stage DO control, KSND maintained high abundance in oligotrophic PTBR, removing 87.88% TN from low-carbon to nitrogen domestic sewage in 180-days.

Evaluation of a novel low-carbon to nitrogen- and temperature-tolerant simultaneously nitrifying-denitrifying bacterium and its use in the treatment of river water.

In this study, a simultaneously heterotrophic nitrifying-aerobic denitrifying bacterium, designated KSND, was newly isolated from a lake wetland. Its removal efficiencies for 160 mg L-1 ammonium, 105 mg L-1 nitrate, and 8.39 mg L-1 nitrite were 86.56%, 74.52%, and 100% in 24 h, with removal rates of 5.77 mg L-1 h-1 for NH4 +-N, 3.26 mg L-1 h-1 for NO3 --N, and 0.35 mg L-1 h-1 for NO2 --N. The bacterium retained ∼63% of its maximal removal rate at 10 °C and 56% of its maximal removal rate at a carbon to nitrogen (C/N) ratio of 4 : 1, with no nitrite accumulation. Gene-specific PCR indicated the absence of the key genes for nitrification and denitrification, encoding hydroxylamine oxidoreductase and nitrite reductase, respectively, suggesting that KSND achieves effective nitrogen removal by another pathway. KSND was used to treat river wastewater by culturing it in a floating bed bioreactor. Ammonia nitrogen decreased significantly from 8.76 mg L-1 initially to 1.87 mg L-1 in 90 days, with no NO3 --N or NO2 --N toxicants, indicating the great potential utility of KSND in future full-scale applications in the treatment of low-C/N wastewater.

In situ monitoring of Shewanella oneidensis MR-1 biofilm growth on gold electrodes by using a Pt microelectrode.

Much attention has been focused on electrochemically active bacteria (EAB) in the application of bioelectrochemical systems (BESs). Studying the EAB biofilm growth mechanism as well as electron transfer mechanism provides a route to upgrade BES performance. But an effective bacterial growth monitoring method on the biofilm scale is still absent in this field. In this work, electrode-attached bacterial biofilms formed by Shewanella oneidensis MR-1 were dynamically monitored through a microelectrode method. For S. oneidensis MR-1, a respiratory electron transport chain is associated with the secretion of riboflavin, severing as the cofactor to the outer membrane c-type cytochromes. The biofilm growth was monitored through adopting riboflavin as an electrochemical probe during the approach of the microelectrode to the biofilm external surface. This method allows in vivo and in situ biofilm monitoring at different growth stages without destructive manipulation. Furthermore, the biofilm growth monitoring results have been proved to be relatively accurate through observation under confocal laser scanning microscopy. We further applied this method to investigate the effects of four environmental factors (the concentrations of dissolved oxygen, sodium lactate, riboflavin as well as the electrode potential) on S. oneidensis MR-1 biofilm development.

Heterogeneous UV/Fenton catalytic degradation of wastewater containing phenol with Fe-Cu-Mn-Y catalyst.

The heterogeneous UV/Fenton process with the appropriate amount of Fe-Mn-Cu-Y as catalyst was developed and various operation conditions for the degradation of phenol were evaluated. The results indicated that by using the heterogeneous UV/Fenton process, the COD(cr) removal rate reached almost 100% for wastewater containing phenol. Compared with the homogeneous process, the developed catalyst could be used at wider pH range in the UV/Fenton process. Comparison of various heterogeneous process showed that heterogeneous UV/Fenton process was best. The heterogeneous UV/Fenton process with Fe-Mn-Cu-Y catalyst is highly efficient in degrading various organic pollutants.