Haijun Ma

Characterization of microbial community and antibiotic resistance genes in activated sludge under tetracycline and sulfamethoxazole selection pressure

To investigate the microbial community characteristics, antibiotic resistance genes (ARGs), and bioreactor effluent quality change under tetracycline (TC) and sulfamethoxazole (SMX) selection pressure, sequencing batch reactors (SBRs) were used with environmentally relevant concentration and high-level of TC and SMX concentrations (0, 5ppb, 50ppb and 10ppm). Chemical oxygen demand (COD) and ammonia nitrogen (NH4(+)N) removals appeared unchanged (p>0.05) with 5 and 50ppb, but decreased significantly with 10ppm (p<0.05). Extracellular polymeric substances (EPS) concentrations increased significantly with increasing TC or SMX concentrations (p<0.05). High-throughput 16S rRNA gene sequencing results suggested that Proteobacteria, Actinobacteria and Bacteroidetes were the three most abundant phyla in sludge samples. The Actinobacteria percentages increased with increasing TC or SMX concentration, while Proteobacteria and Bacteroidetes decreased. The microbial diversity achieved its maximum at 5ppb and decreased with higher concentrations. The total ARGs abundances in sludge increased with addition of TC or SMX, and the higher relative abundances were in the order of sul1>tetG>sul2>tetA>intI1>tetS>tetC. Pearson correlation analysis showed most ARGs (tetA, tetC, tetG, tetK, tetM, sul1) were significantly correlated with intI1 (p<0.01).

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.

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.

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.

Exogenous N -acyl homoserine lactones facilitate microbial adhesion of high ammonia nitrogen wastewater on biocarrier surfaces

Startup of biofilm process triggered by initial adhesion of bacteria is difficult in high ammonia nitrogen wastewater treatment. In this study, the influence of two commonly used N-acyl homoserine lactones (AHLs), N-Hexanoyl-l-homoserine lactone (C6-HSL) and N-Octanoyl-l-homoserine lactone (C8-HSL), on the adhesion of soluble macromolecules and bacteria in four types of high ammonia nitrogen wastewater to surfaces of model biocarriers (i.e. polystyrene, polyamide and polyethylene terephthalate) was investigated by using a quartz crystal microbalance with dissipation (QCM-D) monitoring technology. Results showed that the adhesion was enhanced by the addition of exogenous AHLs and there was more microbial retention attributed by C8-HSL. Greater deposition amount was generally found on PS and better enhanced performances of the adhesion were found on PA surface. Furthermore, viscoelastic film formed under synchronous high-low salinity and organic content and dominant bacteria of real wastewater determined the role of exogenous AHLs. The method of adding moderate amount of exogenous AHLs into bioreactors has important implications for accelerating the startup process treating high ammonia nitrogen wastewater by biofilm process.

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.

Cell membrane characteristics and microbial population distribution of MBBR and IFAS with different dissolved oxygen concentration

This paper investigated the influences of different dissolved oxygen (DO) concentration (0.71-1.32, 2.13-3.02 and 4.31-5.16 mg/L) on cell membrane characteristics and microbial population distribution of moving biofilm reactors. Two representative reactors, i.e., moving bed biofilm reactors and integrated fixed-film activated sludge were operated. Results indicated that both DO concentration of 0.71-1.32 mg/L and 4.31-5.16 mg/L could increase membrane lipid mobile fraction (49.4%-67.4%) of the microbes, however, through prompting the synthesis of branched fatty acids and unsaturated fatty acids, respectively. For the biofilms, the abundance of Bacteroidetes decreased and Actinobacteria increased with the increase of DO levels. The lowest EfOM content and the highest microbial diversities (1.14-1.52) was observed at DO of 2.13-3.02 mg/L. Redundancy analysis showed that changes of DO levels could alter cell membrane properties and bacterial community structures, and subsequently significantly influenced effluent organic matter composition of moving biofilm reactors.

Distribution of endotoxins in full scale pharmaceutical wastewater treatment plants and its relationship with microbial community structure

Endotoxins are potential toxics impacting human health through respiration derived in wastewater treatment plants (WWTPs), yet the formation of endotoxins during wastewater treatment processes is still lacking research. In our study, the distribution of endotoxins and bacterial community structure in the wastewater of three full scale pharmaceutical WWTPs were explored using the limulus amebocyte lysate (LAL) test and MiSeq technique. Results showed that higher endotoxin activities in the influent of Plant A and Plant C (560 and 1140 EU/mL), stemming from the fermentation process, were found compared to that of Plant B (135 EU/mL), coming from the process of chemical synthesis. During the anaerobic treatment and the cyclic activated sludge system (CASS) in the three WWTPs, the endotoxin activity increased, while it declined in the aerobic treatment system. In all bioreactors, the relative abundance of Gram-negative bacteria accounted for 50.0-94.6%. Bacteria with high lipopolysaccharide (LPS) in LAL assay were found at the genus level of Bacteroides, Enterococcus, Desulfovibrio, and Megasphaera.

Effect of mixing intensity on hydrolysis and acidification of sewage sludge in two-stage anaerobic digestion: Characteristics of dissolved organic matter and the key microorganisms

Mixing should be optimized in anaerobic digestion (AD) systems to achieve excellent biomaterials production in the sewage sludge (SS) management in wastewater treatment plant. AD depends on the coordinated activity of hydrolysis, acidification and methanogenesis. However, the effect of mixing intensity on characteristics of hydrolysis and acidification in AD of SS is still poorly understood. This study focused on the mixing intensity (30, 60, 90 and 120 rpm) effect on the characteristics of dissolved organic matter (DOM) and the key microorganisms in the hydrolysis and acidification of SS. Results showed that enhanced hydrolysis and acidification efficiency was obtained at mixing of 90 and 120 rpm (p < 0.05), while the maximum acetic acid (388 ± 21 mg/L) was produced at 90 rpm. Mixing at 90 rpm enhanced the release of protein and polysaccharide as well as humic acid. Further analyses of DOM molecular features revealed that 90 rpm led to the highest molecular diversity and easily biodegradable molecules (lipid and proteins/amino sugars), which contributed to the maximum hydrolysis and acidification efficiency. Firmicutes and Actinobacteria significantly increased with mixing intensity (p<0.05), and Chloroflexi and Fusobacteria were enriched at mixing of 90 rpm, which favored the hydrolysis of SS. The enrichment of Clostridium XI and Clostridium sensu stricto contributed to the acidification of DOM at 90 and 120 rpm. The results of this study can advance our knowledge about mixing intensity effects on the AD systems of SS. This research also showed how increasing mixing intensity to a relatively high speed can enhance the hydrolysis and acidification efficiency of SS.

The biological role of N-acyl-homoserine lactone-based quorum sensing (QS) in EPS production and microbial community assembly during anaerobic granulation process

Although N-acyl-L-homoserine lactone (AHL) based quorum sensing (QS) phenomenon has been observed in mature anaerobic granules, the biological role of AHL-based QS system in anaerobic granulation process remains unexplored. For the first time, a long-term anaerobic bioreactor was operated for 168 days to investigate the biological role of AHL in the granulation process which was divided into three phases (phase I: floccular, phase II: granulation, phase III: maturation). Two different AHLs including C8-HSL and C10-HSL were characterized at nanogram levels. The AHL level was elevated over 20-fold and strongly positively correlated with extracellular polymeric substances (EPS) production and sludge particle size during phase I-II. Exogenous addition of AHL to the floccular sludge also resulted in significantly increased EPS production. Metadata analysis suggested that the granulation process was accompanied by an increase in the abundance of QS-relevant microorganisms. The strong relationships (R > 0.9233, p < 0.01) among AHL concentrations, EPS (except loosely bound EPS), granulation and community variation indicated that AHL-mediated QS played an important role in coordinating community level behaviors associated with granulation, potentially through the regulation of EPS production and composition. This study gives a deep insight into the underlying QS-relevant mechanism of anaerobic granulation process.