Mitigating microbiological risks of potential pathogens carrying antibiotic resistance genes and virulence factors in receiving rivers: Benefits of wastewater treatment plant upgrade

● Abundance of MAGs carrying ARG-VF pairs unchanged in rivers after WWTP upgrade. ● Upgrade of WWTPs significantly reduced diversity of pathogenic genera in rivers. ● Upgrade of WWTPs reduced most VF (ARG) types carried by potential pathogens in rivers. ● Upgrade of WWTPs narrowed the pathogenic host ranges of ARGs and VFs in rivers. Wastewater treatment plants (WWTPs) with additional tertiary ultrafiltration membranes and ozonation treatment can improve water quality in receiving rivers. However, the impacts of WWTP upgrade (WWTP-UP) on pathogens carrying antibiotic resistance genes (ARGs) and virulence factors (VFs) in rivers remain poorly understood. In this study, ARGs, VFs, and their pathogenic hosts were investigated in three rivers impacted by large-scale WWTP-UP. A five-year sampling campaign covered the periods before and after WWTP-UP. Results showed that the abundance of total metagenome-assembled genomes (MAGs) containing both ARGs and VFs in receiving rivers did not decrease substantially after WWTP-UP, but the abundance of MAGs belonging to pathogenic genera that contain both ARGs and VFs (abbreviated as PAVs) declined markedly. Genome-resolved metagenomics further revealed that WWTP-UP not only reduced most types of VFs and ARGs in PAVs, but also effectively eliminated efflux pump and nutritional VFs carried by PAVs in receiving rivers. WWTP-UP narrowed the pathogenic host ranges of ARGs and VFs and mitigated the co-occurrence of ARGs and VFs in receiving rivers. These findings underline the importance of WWTP-UP for the alleviation of pathogens containing both ARGs and VFs in receiving rivers.     

Degradation of extracellular genomic,plasmid DNA and specific antibiotic resistance genes by chlorination

Extracellular DNA structure damaged by chlorination was characterized. Integrity of extracellular ARG genetic information after chlorination was determined. Typical chlorine doses will likely effectively diminish extracellular DNA and ARGs. Plasmid DNA/ARGs were less readily broken down than genomic DNA. The Bioanalyzer methodology effectively documented damage incurred to DNA. There is a need to improve understanding of the effect of chlorine disinfection on antibiotic resistance genes (ARGs) in order to advance relevant drinking water, wastewater, and reuse treatments. However, few studies have explicitly assessed the physical effects on the DNA. Here we examined the effects of free chlorine (1–20 mg Cl₂/L) on extracellular genomic, plasmid DNA and select ARGs. Chlorination was found to decrease the fluorometric signal of extracellular genomic and plasmid DNA (ranging from 0.005 to 0.05 mg/mL) by 70%, relative to a no-chlorine control. Resulting DNA was further subject to a fragment analysis using a Bioanalyzer, indicating that chlorination resulted in fragmentation. Moreover, chlorine also effectively deactivated both chromosomal- and plasmid-borne ARGs, mecA and tetA, respectively. For concentrations >2 mg Cl₂//L × 30 min, chlorine efficiently reduced the qPCR signal when the initial concentration of ARGs was 10⁵ copies/mL or less. Notably, genomic DNA and mecA gene signals were more readily reduced by chlorine than the plasmid-borne tetA gene (by ~2 fold). Based on the results of qPCR with short (~200 bps) and long amplicons (~1200 bps), chlorination could destroy the integrity of ARGs, which likely reduces the possibility of natural transformation. Overall, our findings strongly illustrate that chlorination could be an effective method for inactivating extracellular chromosomal- and plasmid-borne DNA and ARGs.     

Municipal wastewater treatment in China: Development history and future perspectives

The history of China’s municipal wastewater management is revisited. The remaining challenges in wastewater sector in China are identified. New concept municipal wastewater treatment plants are highlighted. An integrated plant of energy, water and fertilizer recovery is envisaged. China has the world’s largest and still growing wastewater sector and water market, thus its future development will have profound influence on the world. The high-speed development of China’s wastewater sector over the past 40 years has forged its global leading treatment capacity and innovation ability. However, many problems were left behind, including underdeveloped sewers and sludge disposal facilities, low sustainability of the treatment processes, questionable wastewater treatment plant (WWTP) effluent discharge standards, and lacking global thinking on harmonious development between wastewater management, human society and the nature. Addressing these challenges calls for fundamental changes in target design, policy and technologies. In this mini-review, we revisit the development history of China’s municipal wastewater management and identify the remaining challenges. Also, we highlight the future needs of sustainable development and exploring China’s own wastewater management path, and outlook the future from several aspects including targets of wastewater management, policies and technologies, especially the new concept WWTP. Furthermore, we envisage the establishment of new-generation WWTPs with the vision of turning WWTP from a site of pollutant removal into a plant of energy, water and fertilizer recovery and an integrated part urban ecology in China.     

Assessment of antibiotic resistance genes in dialysis water treatment processes

• Quantitative global ARGs profile in dialysis water was investigated. • Totally 35 ARGs were found in the dialysis treatment train. • 29 ARGs (highest) were found in carbon filtration effluent. • erm and mtrD-02 occurred in the final effluent. • The effluent was associated with health risks even after RO treatment. Dialysis water is directly related to the safety of hemodialysis patients, thus its quality is generally ensured by a stepwise water purification cascade. To study the effect of water treatment on the presence of antibiotic resistance genes (ARGs) in dialysis water, this study used propidium monoazide (PMA) in conjunction with high throughput quantitative PCR to analyze the diversity and abundance of ARGs found in viable bacteria from water having undergone various water treatment processes. The results indicated the presence of 35 ARGs in the effluents from the different water treatment steps. Twenty-nine ARGs were found in viable bacteria from the effluent following carbon filtration, the highest among all of the treatment processes, and at 6.96 Log (copies/L) the absolute abundance of the cphA gene was the highest. Two resistance genes, erm (36) and mtrD-02, which belong to the resistance categories macrolides-lincosamides-streptogramin B (MLSB) and other/efflux pump, respectively, were detected in the effluent following reverse osmosis treatment. Both of these genes have demonstrated the potential for horizontal gene transfer. These results indicated that the treated effluent from reverse osmosis, the final treatment step in dialysis-water production, was associated with potential health risks.     

Distribution of antibiotic resistance genes and their association with bacteria and viruses in decentralized sewage treatment facilities

• Distribution of ARGs in decentralized sewage facilities were investigated. • Bacitracin-ARGs were most predominant ARGs in rural wastewater. • ARGs were identified in bacterial and viral community. • ARGs of rpoB, drfE, gyrA and parC were both correlated with bacteria and phages. • More attention should be paid to the risk of spreading ARG by phages. The distribution of antibiotic resistance genes (ARGs) has been intensively studied in large-scale wastewater treatment plants and livestock sources. However, small-scale decentralized sewage treatment facilities must also be explored due to their possible direct exposure to residents. In this study, six wastewater treatment facilities in developed rural areas in eastern China were investigated to understand their risks of spreading ARGs. Using metagenomics and network analysis tools, ARGs and bacterial and viral communities were identified in the influent (INF) and effluent (EFF) samples. The dominant ARGs belonged to the bacitracin class, which are different from most of municipal wastewater treatment plants (WWTPs). The dominant hosts of ARGs are Acidovorax in bacterial communities and Prymnesiovirus in viral communities. Furthermore, a positive relationship was found between ARGs and phages. The ARGs significantly correlated with phages were all hosted by specific genera of bacteria, indicating that phages had contributed to the ARG’s proliferation in sewage treatment facilities. Paying significant concern on the possible enhanced risks caused by bacteria, viruses and their related ARGs in decentralized sewage treatment facilities is necessary.     

Presence,dissemination and removal of antibiotic resistant bacteria and antibiotic resistance genes in urban drinking water system: A review

Reviewed the change of ARGs and ARB in full-scale urban drinking water systems. Conventional processes are more promising than BAC process in ARGs removal. Mechanisms of ARGs enrichment and spread in BAC filter and DWDSs are discussed. Raise the need of future research on ARGs and ARB change in building plumbing systems. Antibiotic resistance in aquatic environment has become an important pollution problem worldwide. In recent years, much attention was paid to antibiotic resistance in urban drinking water systems due to its close relationship with the biosafety of drinking water. This review was focused on the mechanisms of antibiotic resistance, as well as the presence, dissemination and removal of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in the urban drinking water system. First, the presence of ARB and ARGs in the drinking water source was discussed. The variation of concentration of ARGs and ARB during coagulation, sedimentation and filtration process were provided subsequently, in which filtration was proved to be a promising technology to remove ARGs. However, biological activated carbon (BAC) process and drinking water distribution systems (DWDSs) could be incubators which promote the antibiotic resistance, due to the enrichment of ARGs and ARB in the biofilms attached to the active carbon and pipe wall. Besides, as for disinfection process, mechanisms of the inactivation of ARB and the promotion of conjugative transfer of ARGs under chlorine, ozone and UV disinfection were described in detail. Here we provide some theoretical support for future researches which aim at antibiotic resistance controlling in drinking water.     

Microplastics in municipal wastewater treatment plants: a case study of Denizli/Turkey

● High amounts of microplastics are released to receiving media from WWTPs. ● The effect of classical treatment processes on MP removal is important. ● MP load in the effluent of WWTPs is important for developing treatment technology. ● Additional physical treatment could help further reduce MP discharge. Plastic particles smaller than 5 mm are microplastics. They are among the significant pollutants that recently attracted attention. Great quantities of microplastics enter the sewage system daily and reach wastewater treatment plants (WWTPs). As a result, WWTPs are potential microplastic sources. Hence, they create a pathway for microplastics to reach aquatic environments with treated wastewater discharge. Studies on microplastic characterization in WWTPs have gained momentum in academia. This study investigates the abundance, size, shape, color, polymer type, and removal efficiencies of microplastics in a municipal wastewater treatment plant (WWTP) in Denizli/Turkey. The results showed that the dominant microplastic shape in wastewater samples was fibers (41.78%–60.77%) in the 100–500 µm (58.57%–80.07%) size range. Most of the microplastics were transparent-white (32.86%–58.93%). The dominant polymer types were polyethylene (54.05%) and polyethylene vinyl acetate (37.84%) in raw wastewater. Furthermore, the microplastic removal efficiencies of the Denizli Central WWTP as a whole and for individual treatment units were evaluated. Although the microplastic pollution removal efficiency of the Denizli Central WWTP was over 95%, the microplastic concentration discharged daily into the receiving environment was considerably high (1.28 × 10¹⁰ MP/d). Thus, Denizli Central WWTP effluents result in a high volume of emissions in terms of microplastic pollution with a significant daily discharge to the Çürüksu Stream.     

Effect of the ultraviolet/chlorine process on microbial community structure,typical pathogens,and antibiotic resistance genes in reclaimed water

• UV/chlorine can effectively remove VBNC pathogens, ARGs and MGEs in reclaimed water. • Microbial community was changed with reduced diversity during UV/chlorine process. • CRBs-carried MGEswere the predominant groups during UV/chlorine process. • No direct co-selection strategy was shared between UV/chlorine and resistome. Urban wastewater contains a wide range of pathogens and antibiotic resistance genes (ARGs), which are a serious concern if reusing treated wastewater. However, few studies have explored the microbial communities in reclaimed water using ultraviolet (UV)/chlorine treatment and assessed the changes of the resistome. This study investigated the occurrence of typical pathogens, ARGs, and bacterial communities in UV/chlorine-treated reclaimed water samples. The numbers of culturable and viable but non-culturable pathogens were effectively reduced to 0 CFU/mL within 1–10 and 10–30 min after UV/chlorine treatment, respectively. Meanwhile, the physicochemical indices of water quality were not affected. UV/chlorine treatment could significantly change the bacterial community structure of reclaimed water, showing a decrease in bacterial abundance and diversity. Chlorine-resistant Acinetobacter and Mycobacterium were the dominant bacterial genera (>50%) after UV/chlorine treatment. Moreover, the number of ARGs and mobile genetic elements (MGEs) decreased with an increase in UV/chlorine exposure. However, eight ARGs and three MGEs were consistently detected in more than three seasons, making these major concerns because of their potential role in the persistence and dissemination of antibiotic resistance. Overall, the results of this study suggest that UV/chlorine treatment can potentially improve the microbiological safety of reclaimed water. And more attention should be paid to the pathogens that are both chlorine-resistant and carry MGEs because of their potential for resistance transmission.     

Metagenomic analysis on resistance genes in water and microplastics from a mariculture system

• Total 174 subtypes of ARGs were detected by metagenomic analysis. • Chloramphenicol resistance genes were the dominant ARGs in water and microplastics. • The abundances of MRGs were much higher than those of ARGs. • Proteobacteria, Bacteroidetes, and Actinobacteria were the dominant phylum. • Microplastics in mariculture system could enrich most of MRGs and some ARGs. Microplastics existing widely in different matrices have been regarded as a reservoir for emerging contaminants. Mariculture systems have been observed to host microplastics and antibiotic resistance genes (ARGs). However, more information on proliferation of ARGs and metal resistance genes (MRGs) in mariculture system at the presence of microplastics is needed. This study used metagenomic analysis to investigate the distribution of ARGs and MRGs in water and microplastics of a typical mariculture pond. Total 18 types including 174 subtypes of ARGs were detected with the total relative abundances of 1.22/1.25 copies per 16S rRNA copy for microplastics/water. Chloramphenicol resistance genes were the dominant ARGs with the abundance of 0.35/0.42 copies per 16S rRNA copy for microplastics/water. Intergron intI1 was dominant gene among 6 detected mobile genetic elements (MGEs) with the abundance of 75.46/68.70 copies per 16S rRNA copy for water/microplastics. Total 9 types including 46 subtypes of MRGs were detected with total abundance of 5.02 × 10²/6.39 × 10² copies per 16S rRNA copy for water/ microplastics while genes resistant to copper and iron served as the dominant MRGs. Proteobacteria, Bacteroidetes, and Actinobacteria accounted for 84.2%/89.5% of total microbial community. ARGs with relatively high abundance were significantly positively related to major genera, MGEs, and MRGs. Microplastics in mariculture system could enrich most of MRGs and some ARGs to serve as potential reservoir for these pollutants. The findings of this study will provide important information on resistance gene pollution at presence of microplastics in the mariculture system for further proposing suitable strategy of environmental management.     

城市污水处理厂微生物气溶胶污染和粒径分布特征

城市污水处理厂微生物气溶胶的污染与控制逐渐成为人们关注的热点问题。为明确城市污水处理厂微生物气溶胶浓度和粒径分布特征,选择鄂尔多斯市某城市污水处理厂的格栅间、配水池、氧化沟、二沉池、污泥脱水间和厂区门口6个功能区为对象,对各功能区产生的异养细菌、真菌和放线菌气溶胶浓度分布、污染特性和微生物粒子粒径分布特性进行研究。结果表明,该污水处理厂不同功能区内异养细菌、真菌和放线菌的浓度分布存在显著性差异,其中二沉池和污泥脱水间产生的异养细菌和放线菌浓度均较高,真菌浓度较高的区域为厂门口和二沉池;根据相关微生物气溶胶污染评价标准,污泥脱水间逸散的异养细菌浓度均达到污染级,二沉池、氧化沟和厂门口逸散的异养细菌浓度达轻微污染级,格栅间和配水池未受污染,而真菌浓度在各个功能区均未受到污染;各功能区产生的异养细菌、真菌和放线菌粒子粒径分布无明显差异,异养细菌、真菌和放线菌粒子粒径分别集中在第1、4级,第3、4级和第1、2、3级,大体上均呈正态分布;各功能区均逸散出一定比例的可以直接吸入人体呼吸道的微生物粒子,可能对人体健康具有一定的潜在风险。研究结果可为城市污水处理厂运行过程中微生物气溶胶的污染控制提供科学依据。     

Characterization of the genes involved in nitrogen cycling in wastewater treatment plants using DNA microarray and most probable number-PCR

To improve nitrogen removal performance of wastewater treatment plants (WWTPs), it is essential to understand the behavior of nitrogen cycling communities, which comprise various microorganisms. This study characterized the quantity and diversity of nitrogen cycling genes in various processes of municipal WWTPs by employing two molecular-based methods:most probable number-polymerase chain reaction (MPN-PCR) and DNA microarray. MPN-PCR analysis revealed that gene quantities were not statistically different among processes, suggesting that conventional activated sludge processes (CAS) are similar to nitrogen removal processes in their ability to retain an adequate population of nitrogen cycling microorganisms. Furthermore, most processes in the WWTPs that were researched shared a pattern:the nirS and the bacterial amoA genes were more abundant than the nirK and archaeal amoA genes, respectively. DNA microarray analysis revealed that several kinds of nitrification and denitrification genes were detected in both CAS and anaerobic-oxic processes (AO), whereas limited genes were detected in nitrogen removal processes. Results of this study suggest that CAS maintains a diverse community of nitrogen cycling microorganisms; moreover, the microbial communities in nitrogen removal processes may be specific.

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Effectiveness of tertiary treatment processes in removing different classes of emerging contaminants from domestic wastewater

● Different advanced treatment processes were tested for ECs removal from wastewater. ● UV radiation showed low to moderate removal for 5 of the 38 micropollutants. ● Among tested membrane processes, nanofiltration showed the better performance. ● The use of PAC achieved high or partially removal for 31 out of the 38 compounds. ● The environmental and economical evaluation of a pilot-scale PAC unit is suggested. In this work, 38 different organic emerging contaminants (ECs), belonging to various chemical classes such as pharmaceuticals (PhCs), endocrine-disrupting chemicals (EDCs), benzotriazoles (BTRs), benzothiazoles (BTHs), and perfluorinated compounds (PFCs), were initially identified and quantified in the biologically treated wastewater collected from Athens’ (Greece) Sewage Treatment Plant (STP). Processes already used in existing STPs such as microfiltration (MF), nanofiltration (NF), ultrafiltration (UF), UV radiation, and powdered activated carbon (PAC) were assessed for ECs’ removal, under the conditions that represent their actual application for disinfection or advanced wastewater treatment. The results indicated that MF removed only one out of the 38 ECs and hence it was selected as pretreatment step for the other processes. UV radiation in the studied conditions showed low to moderate removal for 5 out of the 38 ECs. NF showed better results than UF due to the smaller pore sizes of the filtration system. However, this enhancement was observed mainly for 8 compounds originating from the classes of PhCs and PFCs, while the removal of EDCs was not statistically significant. Among the various studied technologies, PAC stands out due to its capability to sufficiently remove most ECs. In particular, removal rates higher than 70% were observed for 9 compounds, 22 were partially removed, while 7 demonstrated low removal rates. Based on our screening experiments, future research should focus on scaling-up PAC in actual conditions, combining PAC with other processes, and conduct a complete economic and environmental assessment of the treatment.     

Occurrence and migration of microplastics and plasticizers in different wastewater and sludge treatment units in municipal wastewater treatment plant

● Reduce the quantifying MPs time by using Nile red staining. ● The removal rate of MPs and PAEs in wastewater and sludge were investigated. ● MPs and PAEs were firstly analyzed during thermal hydrolysis treatment. ● The removal of PAEs from wastewater and sludge was mainly biodegradation. Microplastics (MPs) and plasticizers, such as phthalate esters (PAEs), were frequently detected in municipal wastewater treatment plants (MWTP). Previous research mainly studied the removal of MPs and PAEs in wastewater. However, the occurrence of MPs and PAEs in the sludge was generally ignored. To comprehensively investigate the occurrence and the migration behaviors of MPs and PAEs in MWTP, a series of representative parameters including the number, size, color, shape of MPs, and the concentrations of PAEs in wastewater and sludge were systematically investigated. In this study, the concentrations of MPs in the influent and effluent were 15.46±0.37 and 0.30±0.14 particles/L. The MP removal efficiency of 98.1% was achieved and about 73.8% of MPs were accumulated in the sludge in the MWTP. The numbers of MPs in the sludge before and after digestion were 4.40±0.14 and 0.31±0.01 particles/g (dry sludge), respectively. Fourier Transform Infrared Spectroscopy (ATR FT-IR) analysis showed that the main types of MPs were polyethylene terephthalate (PET), polypropylene (PP), polyethylene (PE), and polystyrene (PS). Six PAEs, including phthalate (DMP), diethyl phthalate (DEP), diisobutyl phthalate (DIBP), ortho dibutyl phthalate (DBP), butyl benzyl phthalate (BBP), and bis(2-ethyl) hexyl phthalate (DEHP), were detected in the MWTP. The concentrations of total PAEs (ΣPAEs) in the influent and effluent were 76.66 and 6.28 µg/L, respectively. The concentrations of ΣPAEs in the sludge before and after digestion were 152.64 and 31.70 µg/g, respectively. In the process of thermal hydrolysis, the number and size of MPs decreased accompanied by the increase of the plasticizer concentration.     

Influences and mechanisms of nanofullerene on the horizontal transfer of plasmid-encoded antibiotic resistance genes between E. coli strains

• Sub-inhibitory levels of nC₆₀ promote conjugative transfer of ARGs. • nC₆₀ can induce ROS generation, oxidative stress and SOS response. • nC₆₀ can increase cell membrane permeability and alter gene expression. • Results provide evidence of nC₆₀ promoting antibiotic resistance dissemination. The spread and development of antibiotic resistance globally have led to severe public health problems. It has been shown that some non-antibiotic substances can also promote the diffusion and spread of antibiotic resistance genes (ARGs). Nanofullerene (nC₆₀) is a type of nanomaterial widely used around the world, and some studies have discovered both the biological toxicity and environmental toxicity of nC₆₀. In this study, cellular and molecular biology techniques were employed to investigate the influences of nC₆₀ at sub-minimum inhibitory concentrations (sub-MICs) on the conjugation of ARGs between the E. coli strains. Compared with the control group, nC₆₀ significantly increased the conjugation rates of ARGs by 1.32‒10.82 folds within the concentration range of 7.03‒1800 mg/L. This study further explored the mechanism of this phenomenon, finding that sub-MICs of nC₆₀ could induce the production of reactive oxygen species (ROS), trigger SOS-response and oxidative stress, affect the expression of outer membrane proteins (OMPs) genes, increase membrane permeability, and thus promote the occurrence of conjugation. This research enriches our understanding of the environmental toxicity of nC₆₀, raises our risk awareness toward nC₆₀, and may promote the more rational employment of nC₆₀ materials.     

Environmental effects and risk control of antibiotic resistance genes in the organic solid waste aerobic composting system: A review

• ARGs were detected in livestock manure, sludge, food waste and fermentation dregs. • The succession of microbial community is an important factor affecting ARGs. • Horizontal transfer mechanism of ARGs during composting should be further studied. Antibiotic resistance genes (ARGs) have been diffusely detected in several kinds of organic solid waste, such as livestock manure, sludge, antibiotic fermentation residues, and food waste, thus attracting great attention. Aerobic composting, which is an effective, harmless treatment method for organic solid waste to promote recycling, has been identified to also aid in ARG reduction. However, the effect of composting in removing ARGs from organic solid waste has recently become controversial. Thus, this article summarizes and reviews the research on ARGs in relation to composting in the past 5 years. ARGs in organic solid waste could spread in different environmental media, including soil and the atmosphere, which could widen environmental risks. However, the conventional composting technology had limited effect on ARGs removal from organic solid waste. Improved composting processes, such as hyperthermophilic temperature composting, could effectively remove ARGs, and the HGT of ARGs and the microbial communities are identified as vital influencing factors. Currently, during the composting process, ARGs were mainly affected by three response pathways, (I) “Microenvironment-ARGs”; (II) “Microenvironment-microorganisms-ARGs”; (III) “Microorganisms-horizontal gene transfer-ARGs”, respectively. Response pathway II had been studied the most which was believed that microbial community was an important factor affecting ARGs. In response pathway III, mainly believed that MGEs played an important role and paid less attention to eARGs. Further research on the role and impact of eARGs in ARGs may be considered in the future. It aims to provide support for further research on environmental risk control of ARGs in organic solid waste.     

Feedforward control for nitrogen removal in a pilot-scale anaerobic-anoxic-oxic plant for municipal wastewater treatment

To improve the efficiency of nitrogen removal with lower energy consumption, the study of feedforward control was carried out on a pilot-scale anaerobic-anoxic-oxic (AAO) plant for the treatment of municipal wastewater. The effluent qualities of the pilot plant under different control strategies were investigated. The results indicated that the change of external recycle was not a suitable approach to regulate the sludge concentration of plug-flow reactors; adjusting the aeration valve and dissolved oxygen set-point according to ammonia load could overcome the impact of influent fluctuation; and the denitrification potential could be estimated based on the transit time of anoxic zone and the relative content of carbon resource entering the anoxic zone. Simple feedforward control strategies for aeration and internal recycle were subsequently proposed and validated. The nitrogen removal was successfully improved in the pilot plant. The effluent total nitrogen had decreased by 29.9% and was steadily controlled below 15 mg·L^-1. Furthermore, approximately 38% of the energy for aeration had been saved.     

Identifying human-induced influence on microbial community: A comparative study in the effluent-receiving areas in Hangzhou Bay

Microbial compositions showed high differences in two study areas. COD was the key anthropogenic indicator in the coastal wastewater disposal area. Distinctive microbes capable of degrading toxic pollutants were screened. Microbial communities in effluent-receiving areas followed “niche theory”. Microbial community structure is affected by both natural processes and human activities. In coastal area, anthropegenetic activity can usually lead to the discharge of the effluent from wastewater treatment plant (WWTP) to sea, and thus the water quality chronically turns worse and marine ecosystem becomes unhealthy. Microorganisms play key roles in pollutants degradation and ecological restoration; however, there are few studies about how the WWTP effluent disposal influences coastal microbial communities. In this study, sediment samples were collected from two WWTP effluent-receiving areas (abbreviated as JX and SY) in Hangzhou Bay. First, based on the high-throughput sequencing of 16S rRNA gene, microbial community structure was analyzed. Secondly, several statistical analyses were conducted to reveal the microbial community characteristics in response to the effluent disposal. Using PCoA, the significant difference of in microbial community structure was determined between JX and SY; using RDA, water COD and temperature, and sediment available phosphate and ammonia nitrogen were identified as the key environmental factors for the community difference; using LDA effect size analysis, the most distinctive microbes were found and their correlations with environmental factors were investigated; and according to detrended beta-nearest-taxon-index, the sediment microbial communities were found to follow “niche theory”. An interesting and important finding was that in SY that received more and toxic COD, many distinctive microbes were related to the groups that were capable of degrading toxic organic pollutants. This study provides a clear illustration of eco-environmental deterioration under the long-term human pressure from the view of microbial ecology.     

北江河水中抗生素抗性基因污染初步研究（Preliminary Studies on the Pollution Levels of Antibiotic Resistance Genes in the Water of Beijiang River, South China）

环境中细菌的耐药性,尤其是对抗生素的耐药性已成为影响生态环境的重要因素.论文采用抗生素抗性平板法调查了北江河水中四环素、红霉素及磺胺类这3类抗生素耐药性细菌的存在,采用定性PCR及荧光定量PCR方法分别研究了该水域sul1和sul2这2种磺胺类抗生素抗性基因(ARGs)的存在和含量水平.结果表明,所采集北江水域的9个样品中有5个对四环素有耐药性,7个对红霉素有耐药性,8个对磺胺二甲嘧啶有耐药性;定性PCR实验并经基因测序结果证实,5个样品含有sul1,4个样品含有sul2.进一步的PCR定量分析结果显示,7个样品中均检出sul1和sul2磺胺抗性基因,它们与内对照基因16S-rRNA表达量比值分别在10-2.56~10-0.52及10-3.25~10-1.24范围内,该结果显著高于美国科罗拉多州北部河流的研究结果.此外,数据分析也发现,sul1和sul2磺胺抗性基因的含量水平与该区域水中磺胺含量分布具有一定的相关性,表明外源性抗生素对河流的污染是诱导抗性基因的重要因素.     

An extraction-assay system: Evaluation on flavonols in plant resistance to Pb and Cd by supercritical extraction-gas chromatography

In this research, supercritical carbon dioxide extraction (SFE) showed better extraction effect when compared with Solid- liquid extraction (SLE), Soxhlet extraction (SE) and Ultrasonic extraction (UE), not only in the rate but also the time. The comparison among these three extraction modifiers, including acetone, ethanol and methanol demonstrated that ethanol was preferred to SFE due to its high extraction effect and low toxicology. In addition, parameter of SFE, influence of temperature and pressure were investigated, and the best extraction effect was achieved at the optima conditions, temperature of 40°C and the pressure of 35 MPa. Thus, SFE is a highly effective method for flavonols extraction, requiring minimum energy and producing non-toxic byproduct. SFE-GC system is applied for the evaluation on flavonols that plays a key role in plant resistance to heavy metal, with its content and synthetase gene expression significantly increasing in plant when threatened by heavy metal. Besides, results indicated that flavonols can improve plant resistance to oxidative stress by quenching the redundant ROS in matrix.

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