Effects of operational parameters on bacterial communities in Hong Kong and global wastewater treatment plants.

Wastewater treatment plants (WWTPs) are indispensable biotechnology facilities for modern cities and play an essential role in modern urban infrastructure by employing microorganisms to remove pollutants in wastewater, thus protecting public health and the environment. This study conducted a 13-month bacterial community survey of six full-scale WWTPs in Hong Kong with samples of influent, activated sludge (AS), and effluent to explore their synchronism and asynchronism of bacterial community. Besides, we compared AS results of six Hong Kong WWTPs with data from 1,186 AS amplicon data in 269 global WWTPs and a 9-year metagenomic sequencing survey of a Hong Kong WWTP. Our results showed the compositions of bacterial communities varied and the bacterial community structure of AS had obvious differences across Hong Kong WWTPs. The co-occurrence analysis identified 40 pairs of relationships that existed among Hong Kong WWTPs to show solid associations between two species and stochastic processes took large proportions for the bacterial community assembly of six WWTPs. The abundance and distribution of the functional bacteria in worldwide and Hong Kong WWTPs were examined and compared, and we found that ammonia-oxidizing bacteria had more diversity than nitrite-oxidizing bacteria. Besides, Hong Kong WWTPs could make great contributions to the genome mining of microbial dark matter in the global "wanted list." Operational parameters had important effects on OTUs' abundance, such as the temperature to the genera of Tetrasphaera, Gordonia and Nitrospira. All these results obtained from this study can deepen our understanding of the microbial ecology in WWTPs and provide foundations for further studies. IMPORTANCE: Wastewater treatment plants (WWTPs) are an indispensable component of modern cities, as they can remove pollutants in wastewater to prevent anthropogenic activities. Activated sludge (AS) is a fundamental wastewater treatment process and it harbors a highly complex microbial community that forms the main components and contains functional groups. Unveiling "who is there" is a long-term goal of the research on AS microbiology. High-throughput sequencing provides insights into the inventory diversity of microbial communities to an unprecedented level of detail. At present, the analysis of communities in WWTPs usually comes from a specific WWTP and lacks comparisons and verification among different WWTPs. The wide-scale and long-term sampling project and research in this study could help us evaluate the AS community more accurately to find the similarities and different results for different WWTPs in Hong Kong and other regions of the world.

A systematic study of microplastic occurrence in urban water networks of a metropolis.

The release of microplastics from sewage treatment works (STWs) into the oceans around coastal cities is well documented. However, there are fewer studies on the microplastic abundance in stormwater drains and their emissions into the coastal marine environment via sewage and stormwater drainage networks. Here, we comprehensively investigated microplastic abundance in 66 sewage and 18 sludge samples collected from different process stages at three typical STWs and 36 water samples taken from six major stormwater drains during the dry and wet seasons in Hong Kong, which is a metropolitan city in south China. The results showed that microplastics were detected in all the sewage and stormwater samples, with the abundance ranging from 0.07 to 91.9 and from 0.4 to 36.48 particles/L, respectively, and in all the sludge samples with the abundance ranging from 167 to 936 particles/g (d. w.). There were no significant seasonal variations in the microplastic abundance across all samples of sewage, sludge, and stormwater. For both waterborne sample types, a smaller size (0.02-0.3 mm) and fiber shape were the dominant characteristics of the microplastics. Polyethylene terephthalate (PET) and polypropylene (PP) were the most abundant polymer types in the sewage samples, while polyethylene (PE), PET, PP, and PE-PP copolymer were the most abundant polymer types in the stormwater samples. The estimated range of total daily microplastic loads in the effluent from STWs in Hong Kong is estimated to be 4.48 × 109 - 2.68 × 1010 particles/day, demonstrating that STWs are major pathways of microplastics in coastal environments despite the high removal percentage of microplastics in sewage treatment processes examined. This is the first comprehensive study on microplastics in the urban waters of a coastal metropolis. However, further studies on other coastal cities will enable an accurate estimation of the microplastic contribution of stormwater drains to the world's oceans.

Quantification of SARS-CoV-2 RNA in wastewater treatment plants mirrors the pandemic trend in Hong Kong.

Wastewater-based epidemiology (WBE) for the SARS-CoV-2 virus in wastewater treatment plants (WWTPs) has emerged as a cost-effective and unbiased tool for population-level testing in the community. In the present study, we conducted a 6-month wastewater monitoring campaign from three WWTPs of different flow rates and catchment area characteristics, which serve 28 % (2.1 million people) of Hong Kong residents in total. Wastewater samples collected daily or every other day were concentrated using ultracentrifugation and the SARS-CoV-2 virus RNA in the supernatant was detected using the N1 and E primer sets. The results showed significant correlations between the virus concentration and the number of daily new cases in corresponding catchment areas of the three WWTPs when using 7-day moving average values (Kendall's tau-b value: 0.227-0.608, p < 0.001). SARS-CoV-2 virus concentration was normalized to a fecal indicator using PMMoV concentration and daily flow rates, but the normalization did not enhance the correlation. The key factors contributing to the correlation were also evaluated, including the sampling frequency, testing methods, and smoothing days. This study demonstrates the applicability of wastewater surveillance to monitor overall SARS-CoV-2 pandemic dynamics in a densely populated city like Hong Kong, and provides a large-scale longitudinal reference for the establishment of the long-term sentinel surveillance in WWTPs for WBE of pathogens which could be combined into a city-wide public health observatory.

Fate of bisphenol A, perfluorooctanoic acid and perfluorooctanesulfonate in two different types of sewage treatment works in Hong Kong.

This study aimed at investigating the removal efficiencies of perfluorooctanoate (PFOA), perfluorooctane sulfonate (PFOS) and bisphenol A (BPA) of two major sewage treatment works in Hong Kong: Stonecutters Island STW (SCISTW) which adopts chemically enhanced primary treatment (CEPT) process and Sha Tin STW (STSTW) which employs biological treatment. Liquid portion (LP) and particulate matter (PM) of liquid sewage samples and sludge samples were collected and followed by liquid chromatograph system. It was found that BPA (44.6 ± 35.1%) generally achieved higher TRE than the two other chemicals (PFOS (-18.8 ± 34.8%) and PFOA (-104 ± 86.8%)) in STSTW (p < 0.05). Most of the PFOA, PFOS and BPA was discharged through final effluent (PFOA: 95.6 ± 1.00% and 94.5 ± 3.13%; PFOS: 77.7 ± 1.48% and 72.6 ± 6.07%; BPA: 99.2 ± 0.950% and 92.8 ± 7.25%, respectively) rather than stored in the sludge/cake (PFOA: 4.45 ± 1.00% and 5.47 ± 3.13%, PFOS: 22.3 ± 1.48% and 27.4 ± 6.07%, BPA: 0.844 ± 0.950% and 7.20 ± 7.25%, respectively). After the sewage purification process, the two STW released considerable amounts of PFOA, PFOS and BPA in the final effluent (PFOA: 0.638 ± 0.227 kg/year; PFOS: 0.409 ± 0.126 kg/year; BPA: 10.4 ± 3.83 kg/year in STSTW; PFOA: 3.08 ± 1.415 kg/year; PFOS: 2.13 ± 0.452 kg/year; BPA: 714 ± 768 kg/year in SCISTW) and in the sludge (PFOA: 0.0360 ± 0.0250 kg/year; PFOS: 0.149 ± 0.00100 kg/year; BPA: 1.09 ± 1.47 kg/year in STSTW; PFOA: 0.139 ± 0.0670 kg/year; PFOS: 0.606 ± 0.0780 kg/year; BPA: 3.05 ± 3.95 kg/year in SCISTW). This study may help to provide crucial information for further development of municipal sewage system in treating synthetic emerging chemicals.

Profiles and removal efficiency of polycyclic aromatic hydrocarbons by two different types of sewage treatment plants in Hong Kong.

Sewage discharge could be a major source of polycyclic aromatic hydrocarbons (PAHs) in the coastal waters. Stonecutters Island and Shatin Sewage Treatment Works (SCISTW and STSTW) in Hong Kong, adopted chemically enhanced primary treatment and biological treatment, respectively. This study aimed at (1) determining the removal efficiencies of PAHs, (2) comparing the capabilities in removing PAHs, and (3) characterizing the profile of each individual PAHs, in the two sewage treatment plants (STPs). Quantification of 16 PAHs was conducted by a Gas Chromatography. The concentrations of total PAHs decreased gradually along the treatment processes (from 301±255 and 307±217ng/L to 14.9±12.1 and 63.3±54.1ng/L in STSTW and SCISTW, respectively). It was noted that STSTW was more capable in removing total PAHs than SCISTW with average total removal efficiency 94.4%±4.12% vs. 79.2%±7.48% (p<0.05). The removal of PAHs was probably due to sorption in particular matter, confirmed by the higher distribution coefficient of individual and total PAHs in solid samples (dewatered sludge contained 92.5% and 74.7% of total PAHs in SCISTW and STSTW, respectively) than liquid samples (final effluent-total contained 7.53% and 25.3% of total PAHs in STSTW and SCISTW, respectively). Despite the impressive capability of STSTW and SCISTW in removing PAHs, there was still a considerable amount of total PAHs (1.85 and 39.3kg/year, respectively for the two STPs) being discharged into Hong Kong coastal waters, which would be an environmental concern.

Linking Microbial Community, Environmental Variables, and Methanogenesis in Anaerobic Biogas Digesters of Chemically Enhanced Primary Treatment Sludge.

Understanding the influences of biotic and abiotic factors on microbial community structure and methanogenesis are important for its engineering and ecological significance. In this study, four biogas digesters were supplied with the same inoculum and feeding sludge but operated at different sludge retention time (7 to 16 days) and organic loading rates for 90 days to determine the relative influence of biotic and environmental factors on the microbial community assembly and methanogenic performance. Despite different operational parameters, all digester communities were dominated by Bacteroidales, Clostridiales, and Thermotogales and followed the same trend of population dynamics over time. Network and multivariate analyses suggest that deterministic factors, including microbial competition (involving Bacteroidales spp.), niche differentiation (e.g., within Clostridiales spp.), and periodic microbial immigration (from feed sludge), are the key drivers of microbial community assembly and dynamics. A yet-to-be-cultured phylotype of Bacteroidales (GenBank ID: GU389558.1 ) is implicated as a strong competitor for carbohydrates. Moreover, biogas-producing rate and methane content were significantly related with the abundances of functional populations rather than any operational or physicochemical parameter, revealing microbiological mediation of methanogenesis. Combined, this study enriches our understandings of biological and environmental drivers of microbial community assembly and performance in anaerobic digesters.

Anaerobic digestion of chemically enhanced primary treatment (CEPT) sludge and the microbial community structure.

The effectiveness and treatment conditions of FeCl3- and AlCl3-coagulated municipal sewage sludge from chemically enhanced primary treatment (CEPT) using anaerobic digestion (AD) and the structure of microbial community were investigated. The results based on 297 measurements under different operational conditions demonstrate good average AD performance of CEPT sludge, that is, percent volatile solid reduction of 58 %, specific biogas production (or biogas yield) of 0.92 m(3)/kg volatile solids (VS) destroyed, and methane content of 65.4 %. FeCl3 dosing, organic loading rate, temperature, and hydraulic retention time all significantly affected AD performance. FeCl3 dosing greatly improved specific methane production (methane yield) by 38-54 % and significantly reduced hydrogen sulfide (H2S) content in biogas (from up to 13,250 to <200 ppm), contributing to higher methane recovery and simplified biogas cleaning for power generation. Metagenomic analysis suggested that anaerobic digesters of both CEPT sludge and combined primary and secondary sludge were dominated by Bacteroidetes, Proteobacteria, Firmicutes, Actinobacteria, Thermotogae, and Chloroflexi. However, Methanomicrobia methanogens were better enriched in the anaerobic digesters of CEPT sludge than in the combined sludge. Further, different sources of CEPT sludge with various chemical properties nurtured shared and unique microbial community composition. Combined, this study supports AD as an efficient technology for CEPT sludge treatment and poses first insights into the microbial community structure.

Profiles and removal efficiency of polybrominated diphenyl ethers by two different types of sewage treatment work in Hong Kong.

This study was to investigate removal efficiencies and profiles of 14 polybrominated diphenyl ether (PBDE) congeners by two different types of sewage treatment work (STW) in Hong Kong: Stonecutters Island STW (SCISTW) which uses chemically enhanced primary treatment (CEPT) process and Sha Tin STW (STSTW) which adopts biological treatment. The results indicated that both SCISTW and STSTW had a high total removal efficiency for BDE-47, BDE-99 BDE-209 and total PBDEs (SCISTW: 71.6 ± 15.8, 84.7 ± 12.3, 96.0 ± 2.62 and 87.4 ± 8.02%, respectively; STSTW: 74.8 ± 9.5, 90.7 ± 9.14, 96.2 ± 2.41 and 89.3 ± 2.62%, respectively) and PBDEs were chiefly removed by sorption. However, the profile of PBDEs demonstrated that the relative proportions of BDE-28 and BDE-47 in total PBDEs markedly increased, while that of BDE-209 decreased in the effluent samples of the two sewage treatment works, especially in STSTW. The percentage of BDE-209 in total PBDEs in effluent (49.3%) of SCISTW was 21.2% lower than that in influent (70.5%), and the percentage of BDE-209 in total PBDEs in effluent (13.8%) of STSTW was 34.1% reduced from influent (47.9%). Despite overall removal, the percentage of BDE-47 in total PBDEs in effluent (17.6%) of SCISTW was 6.85% higher than that in influent (10.7%), and the percentage of BDE-47 in total PBDEs in effluent (33.5%) of STSTW was 18.1% increased from influent (16.8%). The increase in proportion of BDE-47 in the effluent might raise environmental and public health concerns. Our study is a first attempt in reporting the PBDE congener profiles in different phases of sewage treatment.

Pilot trial study of a compact macro-filtration membrane bioreactor process for saline wastewater treatment.

Conventional membrane bioreactor (MBR) systems have increasingly been studied in recent decades. However, their applications have been limited due to their drawbacks such as low flux, membrane fouling, and high operating cost. In this study, a compact macro-filtration MBR (MfMBR) process was developed by using a large pore size membrane to mitigate the membrane fouling problem. A pilot trial of MfMBR process was set up and operated to treat 10 m(3)/day of saline wastewater within 4 h. The system was operated under an average permeate flux of 13.1 m(3)/(m(2)·day) for 74 days. The average total suspended solids, total chemical oxygen demand, biological oxygen demand, total Kjeldahl nitrogen, and total nitrogen removal efficiencies achieved were 94.3, 83.1, 98.0, 93.1, and 63.3%, respectively, during steady-state operation. The confocal laser scanning microscopy image indicated that the backwash could effectively remove the bio-cake and dead bacteria. Thus, the results showed that the MfMBR process, which is essentially a primary wastewater treatment process, had the potential to yield the same high quality effluent standards as the secondary treatment process; thereby suggesting that it could be used as an option when the economic budget and/or land space is limited.

Metagenomic analysis of sludge from full-scale anaerobic digesters operated in municipal wastewater treatment plants.

This study applied Illumina high-throughput sequencing to explore the microbial communities and functions in anaerobic digestion sludge (ADS) from two wastewater treatment plants based on a metagenomic view. Taxonomic analysis using SILVA SSU database indicated that Proteobacteria (9.52-13.50 %), Bacteroidetes (7.18 %-10.65 %) and Firmicutes (7.53 %-9.46 %) were the most abundant phyla in the ADS. Differences of microbial communities between the two types of ADS were identified. Genera of Methanosaeta and Methanosarcina were the major methanogens. Functional analysis by SEED subsystems showed that the basic metabolic functions of metagenomes in the four ADS samples had no significant difference among them, but they were different from other microbial communities from activated sludge, human faeces, ocean and soil. Abundances of genes in methanogenesis pathway were also quantified using a methanogenesis genes database extracted from KEGG. Results showed that acetotrophic was the major methanogenic pathway in the anaerobic sludge digestion.