Haidong Hu

Concentration, composition, bioavailability, and N-nitrosodimethylamine formation potential of particulate and dissolved organic nitrogen in wastewater effluents: A comparative study

Wastewater-derived organic nitrogen (org-N) can act as both nutrients and carcinogenic nitrogenous disinfection byproduct precursors. In this study, the concentration, composition, bioavailability, and N-nitrosodimethylamine (NDMA) formation potential of particulate organic nitrogen (PON) from three different municipal wastewater treatment plants were characterized and compared with that of effluent dissolved organic nitrogen (DON). The average effluent PON and DON concentrations ranged from 0.09 to 0.55mgN/L and from 0.91 to 1.88mgN/L, respectively. According to principal component analysis, org-N composition and characterization differed in PON and DON samples (n=20). Compared with DON, PON tended to be enriched in protein and nucleic acids, and showed a more proteinaceous character. Composition of org-N functional groups estimated from the X-ray photoelectron spectroscopy N 1s spectra indicate no significant differences in the molecular weight distribution of the protein-like materials between PON and DON. Moreover, PON exhibited a significantly higher bioavailability (61.0±13.3%) compared to DON (38.5±12.4%, p˂0.05, t-test) and a significantly higher NDMA yields (791.4±404.0ng/mg-N) compared to DON (374.8±62.5ng/mg-N, p˂0.05, t-test). Accordingly, PON contributed to approximately 12.3-41.7% of the total bioavailable org-N and 22.0-38.4% of the total NDMA precursors in wastewater effluents. Thus, the potential adverse effects of PON on wastewater discharge and reuse applications should not be overlooked, even though it only accounted for 7.4-26.8% of the total effluent org-N.

Comparative study of activated sludge with different individual nitrogen sources at a low temperature: Effluent dissolved organic nitrogen compositions, metagenomic and microbial community

The objective of this study was to explore nitrogen removal, especially effluent dissolved organic nitrogen (DON) composition, relative genes and microbial community structures with four individual nitrogen sources at 5°C. Results show that effluent DON did not have dependent relationship with the TN removal rate (urea>ammonia chloride>L-Alanine>D-Alanine). With the same influent TN, the highest effluent DON was formed with urea; the lowest DON was fed with ammonia chloride. The main DON composition was the product of cell metabolism excluding urea, rather than the original substrate. Glutamic acid synthesizing process was of great importance to DON accumulation at 5°C. The nitrogen source type was important to the diversity and heterogeneity of the nitrogen removal genes. Bacterial population structure using redundancy analysis (RDA) showed Simplicispira occupied a higher abundance remarkably in the reactors feeding with urea, and Dyadobacter occupied higher feeding with l-Alanine.

The diversity, distribution and function of N-acyl-homoserine lactone (AHL) in industrial anaerobic granular sludge

Although AHL-mediated quorum sensing (QS) signaling has been proved to be ecologically important in biofilm formation and aerobic granulation process, the biological role of AHL in anaerobic granule has not been experimentally investigated. In this paper, we explored the AHL level in 10 full-scale industrial anaerobic granular bioreactors and detected a total of 4 kinds of AHLs. C8-HSL and C10-HSL were indicated to be the potentially universal QS signal molecules in anaerobic granules and involved in extracellular polymeric substance (EPS) production and granulation process. The add-back experiments further supported the hypothesis that C8-HSL and C10-HSL might play an important role in facilitating anaerobic granulation through regulation of EPS synthesis. 12 Microorganisms might be regulated by AHL to play an important role in EPS production. This study provides a foundation for exploring the function of AHL in anaerobic granular systems, which may shed light on advanced AHL-based anaerobic granulation strategy.

Estimation of spatial distribution of quorum sensing signaling in sequencing batch biofilm reactor (SBBR) biofilms

Quorum sensing (QS) signaling, plays a significant role in regulating formation of biofilms in the nature; however, little information about the occurrence and distribution of quorum sensing molecular in the biofilm of carriers has been reported. In this study, distribution of QS signaling molecules (the acylated homoserine lactones-AHLs, and AI-2), extracellular polymeric substances (EPS) and the mechanical properties in sequencing batch biofilm reactor (SBBR) biofilms have been investigated. Using increased centrifugal force, the biofilms were detached into different fractions. The AHLs ranged from 5.2ng/g to 98.3ng/g in different fractions of biofilms, and N-decanoyl-dl-homoserine lactone (C10-HSL) and N-dodecanoyl-dl-homoserine lactone (C12-HSL) in the biofilms obtained at various centrifugal forces displayed significant differences (p<0.01). Interspecies communication signal autoinducer-2(AI-2) in the biofilms ranged from 79.2ng/g to 98.3ng/g. Soluble EPS and loosely bound EPS content in the different fractions of biofilms displayed significant positive relationship with the distribution of C12-HSL (r=0.86, p<0.05). Furthermore, 49.62% of bacteria in the biofilms were positively related with AHLs with 22.76% was significantly positively (p<0.05) related with AHLs. Biofilm adhesion and compliance was the strongest in the tightly-bound biofilm, the weakest in the supernatant/surface biofilm, which was in accordance with the distribution of C12 HSL(r=0.77, p<0.05) and C10-HSL(r=0.75, p<0.05), respectively. This study addressed on better understanding of possible methods for the improvement of wastewater bio-treatment through biofilm application.

Impact of selected non-steroidal anti-inflammatory pharmaceuticals on microbial community assembly and activity in sequencing batch reactors

This study covers three widely detected non-steroidal anti-inflammatory pharmaceuticals (NSAIDs), diclofenac (DCF), ibuprofen (IBP) and naproxen (NPX), as NSAIDs pollutants. The objective is to evaluate the impact of NSAIDs at their environmental concentrations on microbial community assembly and activity. The exposure experiments were conducted under three conditions (5 μg L-1 DCF, 5 μg L-1 DCF+5 μg L-1 IBP and 5 μg L-1 DCF+5 μg L-1 IBP+ 5 μg L-1 NPX) in sequencing batch reactors (SBRs) for 130 days. Removals of COD and NH4+-N were not affected but total nitrogen (TN) removal decreased. IBP and NPX had the high removal efficiencies (79.96% to 85.64%), whereas DCF was more persistent (57.24% to 64.12%). In addition, the decreased removals of TN remained the same under the three conditions (p > 0.05). The results of oxidizing enzyme activities, live cell percentages and extracellular polymeric substances (EPS) indicated that NSAIDs damaged the cell walls or microorganisms and the mixtures of the three NSAIDs increased the toxicity. The increased Shannon-Wiener diversity index suggested that bacterial diversity was increased with the addition of selected NSAIDs. Bacterial ribosomal RNA small subunit (16S) gene sequencing results indicated that Actinobacteria and Bacteroidetes were enriched, while Micropruina and Nakamurella decreased with the addition of NSAIDs. The enrichment of Actinobacteria and Bacteroidetes indicated that both of them might have the ability to degrade NSAIDs and thereby could adapt well with the presence of NSAIDs.

Determination and Variation of Core Bacterial Community in a Two-Stage Full-Scale Anaerobic Reactor Treating High-Strength Pharmaceutical Wastewater

Knowledge on the functional characteristics and temporal variation of anaerobic bacterial populations is important for better understanding of the microbial process of two-stage anaerobic reactors. However, owing to the high diversity of anaerobic bacteria, close attention should be prioritized to the frequently abundant bacteria that were defined as core bacteria and putatively functionally important. In this study, using MiSeq sequencing technology, the core bacterial community of 98 operational taxonomic units (OTUs) was determined in a two-stage upflow blanket filter reactor treating pharmaceutical wastewater. The core bacterial community accounted for 61.66% of the total sequences and accurately predicted the sample location in the principal coordinates analysis scatter plot as the total bacterial OTUs did. The core bacterial community in the first-stage (FS) and second-stage (SS) reactors were generally distinct, in that the FS core bacterial community was indicated to be more related to a higher-level fermentation process, and the SS core bacterial community contained more microbes in syntrophic cooperation with methanogens. Moreover, the different responses of the FS and SS core bacterial communities to the temperature shock and influent disturbance caused by solid contamination were fully investigated. Co-occurring analysis at the Order level implied that Bacteroidales, Selenomonadales, Anaerolineales, Syneristales, and Thermotogales might play key roles in anaerobic digestion due to their high abundance and tight correlation with other microbes. These findings advance our knowledge about the core bacterial community and its temporal variability for future comparative research and improvement of the two-stage anaerobic system operation.

Nitrogen loading effects on nitrification and denitrification with functional gene quantity/transcription analysis in biochar packed reactors at 5 °C

This study investigated the nitrogen transformation rates of different nitrogen-loading (20, 30, and 50 mg TN/L) biochar packed reactors (C:N:P = 100:5:1) within 125 days at 5 °C. The results showed that high nitrogen loading resulted in an NH4+ (TN) removal efficiency decline from 98% (57%) to 83% (29%), with biochar yielding a higher NH4+, TN and DON removal rate than conventional activated sludge. Moreover, all biochar packed reactors realized a quick start-up by dropping in temperature stage by stage, and the effluent dissolved organic nitrogen (DON) concentrations of R20, R30, and R50 were 0.44 ± 0.18, 0.85 ± 0.35, and 0.66 ± 0.26 mg/L, respectively. The nirS/amoA, nxrA/amoA, and amoA/(narG + napA) were deemed to be the markers of ammonium oxidation rate (SAOR), specific nitrite oxidation rate (SNOR), and specific nitrate reduction rate (SNRR), respectively. Compared with functional gene quantity data, transcription data (mRNA) introduced into stepwise regression analyses agreed well with nitrogen transformation rates. High nitrogen loading also resulted in the cell viability decreased in R50. Nitrogen loadings and operation time both led to a significant variation in cell membrane composition, and unsaturated fatty acids (UFAs) significantly increased in R30 (46.49%) and R50 (36.34%). High-throughput sequencing revealed that nitrogen loadings increased the abundance of nitrifying bacteria (e.g., Nitrospira) and reduced the abundance of denitrifying bacteria (e.g., Nakamurella, Thermomonas, and Zoogloea) through linear discriminant analysis (LDA).

Insight into the characteristics, removal, and toxicity of effluent organic matter from a pharmaceutical wastewater treatment plant during catalytic ozonation

Changes in the characteristics, removal efficiency, and toxicity of pharmaceutical effluent organic matter (EfOM) after catalytic ozonation were investigated in this study. After a 90-min treatment with a catalytic ozonation process (COP) in the presence of MnO2 ceramsite, the total organic carbon (TOC), UV254, colority, protein, and humic acid removal rates were 13.24%, 60.83%, 85.42%, 29.36% and 74.19%, respectively. The polysaccharide content increased by 12.73 mg/L during the COP for reaction times between 0 and ~50 min and decreased by 6.97 mg/L between 50 and ~90 min. Furthermore, 64.44% of the total colority was detected in the hydrophobic organic matter (HOM) fraction, and after the COP, and 88.69% of the colority in the HOM was eliminated. Meanwhile, only 59.18% of the colority in the hydrophilic organic matter (HIM) fraction was removed. GC-MS analysis showed that 38 organic pollutant species were completely removed, 8 were partially removed, and 7 were generated. After 90 min of COP treatment, the pharmaceutical EfOM toxicity was effectively reduced based on the higher incubation and lower mortality rates.

Effect of Solids Retention Time on Effluent Dissolved Organic Nitrogen in the Activated Sludge Process: Studies on Bioavailability, Fluorescent Components, and Molecular Characteristics

Wastewater-derived dissolved organic nitrogen (DON) should be minimized by municipal wastewater treatment plants (MWWTPs) to reduce its potential impact on receiving waters. Solids retention time (SRT) is a key control parameter for the activated sludge (AS) process; however, knowledge of its impact on effluent DON is limited. This study investigated the effect of SRT on the bioavailability, fluorescent components, and molecular characteristics of effluent DON in the AS process. Four lab-scale AS reactors were operated in parallel at different SRTs (5, 13, 26, and 40 days) for treatment of primary treated wastewater collected from an MWWTP. Results showed the positive effect of prolonged SRT on DON removal. AS reactors during longer SRTs, however, cannot sequester the bioavailable DON (ABDON) and occasionally contribute to greater amounts of ABDON in the effluents. Consequently, effluent DON bioavailability increased with SRT ( R2 = 0.619, p < 0.05, ANOVA). Analysis of effluent DON fluorescent components and molecular characteristics indicated that the high effluent DON bioavailability observed at long SRTs is contributed by the production of microbially derived nitrogenous organics. The results presented herein indicate that operating an AS process with a longer SRT cannot control the DON forms that readily stimulate algal growth.

Removal Characteristics of Dissolved Organic Nitrogen and Its Bioavailable Portion in a Postdenitrifying Biofilter: Effect of the C/N Ratio

Addition of external carbon sources to postdenitrification biofilters (DNFs) is frequently used in municipal wastewater treatment plants to enhance dissolved inorganic nitrogen removal. However, little is known about its influence on the removal of dissolved organic nitrogen (DON). This study investigated the effect of the carbon-to-nitrogen (C/N) ratio (3, 4, 5, and 6) on the removal characteristics of DON and bioavailable DON (ABDON) in the pilot-scale DNFs treating real secondary effluent. Results showed that DNFs effluent DON accounted for 31.2-39.8% of the effluent total nitrogen. The maximum effluent DON and ABDON concentrations both occurred in DNF operated at a C/N ratio of 3. There was no significant difference in effluent DON concentrations in DNFs at C/N ratios of 4, 5, and 6; however, effluent ABDON and DON bioavailability significantly decreased with C/N ratios (p < 0.05, t-test). According to the chemical composition analysis, effluent DON at high C/N ratios tends to contain less % molecular weight < 1 kDa nitrogenous organic compounds and proteins/amino sugars-like nitrogenous organic formulas, which is likely responsible for its low bioavailability. Overall, this study indicates the benefit of a high C/N ratio during the DNF process in terms of controlling the DON forms that readily stimulate algal growth.