污水处理厂中磺胺类抗生素、抗性菌、抗性基因的特性

抗生素滥用引起抗性菌的扩散和抗生素抗性基因的污染已成为严重的公共卫生安全隐患。从某污水处理厂出水中分离磺胺类抗性菌,检测其对抗生素耐受性及磺胺类抗生素浓度、抗性基因(antibiotic resistance genes,ARGs)的含量。结果表明水中磺胺类抗生素检测出5种磺胺类抗生素,但浓度较低,3种磺胺类抗生素未检出;磺胺类抗性菌含量为3.18×102~2.24×104CFU/m L,分离的抗性菌对9种抗生素具有抗性,最高耐受力为512 mg/m L,其中,对氯霉素耐受能力最强,对青霉素、氨苄青霉素、头孢氨苄、环丙沙星、庆大霉素、阿奇霉素耐受力次之,对利福平耐受力最弱;抗性基因的绝对含量较低,其中最高含量sul1为105.648~106.956。该研究为STPs抗生素污染的风险评估与抗生素抗性基因污染控制提供了基础数据支持。     

无锡市污水处理厂抗生素抗性菌分布与去除特性研究

近年来,污水处理厂已成为环境中抗生素抗性菌的重要来源.本研究利用传统的异养菌培养法对无锡市6座污水处理厂进出水中的5种抗生素(氨苄霉素、红霉素、四环素、卡那霉素、环丙沙星)抗性菌进行调查与分析,并对比了在不同工艺和不同季节条件下抗性菌的去除效果.结果表明,5种抗性菌在污水处理厂进出水中均可检出,进水中抗性菌浓度为103~10~5CFU·m L~(-1),出水中抗性菌浓度为10~2~10~4CFU·m L~(-1),其中,氨苄霉素抗性菌最多,占总异养菌的比例超过50%.污水处理工艺没有选择性去除抗性菌的效果,出水中抗性菌占总异养菌的比例相对于进水有增加趋势.相比其它传统工艺(氧化沟、SBR等),MBR工艺对去除氨苄霉素、红霉素抗性菌表现出一定优势;夏季目标污水处理厂对抗性菌的去除量略多于冬季,且只有环丙沙星抗性菌的去除量表现出显著的季节性差异(p0.05).     

城市污水处理厂四环素抗性菌的筛选及其抗性基因分析

城市污水处理厂中抗生素抗性菌的富集和扩散对受纳水体及公共卫生安全造成严重的威胁.本研究从上海某城市污水处理厂活性污泥中筛选得到一株四环素抗性菌株TC-1,基于形态特征、生理生化特性和16S r DNA结果鉴定,菌株TC-1属于节杆菌属(Arthrobacter sp.).对节杆菌属菌株的全基因组分析表明,菌株TC-1含有多种抗生素抗性基因,包括四环素、氯霉素、大环内酯类和青霉素抗性基因.对比节杆菌属抗生素抗性基因的基因簇分析表明,这些抗性基因在节杆菌属中具有普遍存在性,表明其来源于相关菌株的遗传而非单个菌株的偶然获得.鉴于节杆菌属是潜在的病原性微生物,该结论可为废水生物处理系统中含抗生素抗性基因的致病微生物环境行为和风险评估提供参考.     

污水处理厂削减耐药菌与抗性基因的研究进展

长期滥用抗生素导致细菌耐药性增强,并使抗性广泛传播.污水处理厂既是耐药菌(antibiotic resistance bacteria,ARB)与抗性基因(antibiotic resistance gene,ARG)的储存库,排放的污水与污泥是向自然环境中传播抗性的重要污染源,也是削减ARB和ARG及控制抗性传播的重要环节.本文总结了天然水体中的耐药菌和抗性基因污染现状,分析了近年来耐药菌与抗性基因在污水/污泥处理过程中的转归与去除方面的研究进展,同时对将来的重点研究方向提出展望,以期为今后耐药菌和抗性基因的污染控制提供参考.     

污水处理厂二级出水中总异养菌群对6种抗生素的耐受性研究

以北京市2座污水处理厂二级出水为研究对象,通过考察总异养菌群、抗性菌比例、浓度及抗生素对细菌的半抑制浓度,研究了二级出水中一般细菌对青霉素、氨苄青霉素、头孢氨苄、氯霉素、四环素和利福平6种抗生素在不同浓度下的耐受性.结果表明,2座污水处理厂出水中青霉素、氨苄青霉素、头孢氨苄和氯霉素抗性菌比例较四环素和利福平高.当抗生素浓度为32mg.L-1时,污水处理厂G二级出水中头孢氨苄抗性菌比例最高为59%,而污水处理厂Q二级出水中氯霉素抗性菌比例最高为44%.头孢氨苄抗性菌在污水处理厂G、Q出水中的浓度分别高达4.0×103 CFU.mL-1和3.5×104 CFU.mL-1,而氯霉素抗性菌浓度分别高达4.9×102 CFU.mL-1和4.6×104 CFU.mL-1.污水处理厂G中异养菌对头孢氨苄的耐受能力最强,其半抑制浓度〉32 mg.L-1;污水处理厂Q中,异养菌对氯霉素的耐受能力最强,其半抑制浓度为23.1 mg.L-1.污水处理厂二级出水中部分抗生素抗性菌污染严重,且稳定存在于低浓度抗生素的处理出水.     

污水处理厂消毒技术对抗生素抗性菌的强化去除

基于消毒技术对污水处理厂出水中总异养菌(total heterotrophic bacteria,HPC)及5种抗生素抗性菌(antibiotic resistant bacteria,ARB):氨苄霉素抗性菌(AMP)、红霉素抗性菌(ERY)、四环素抗性菌(TET)、卡那霉素抗性菌(KAN)、环丙沙星抗性菌(CIP)的去除情况研究,分析消毒技术对ARB的强化去除效果.结果表明,实际污水处理厂的紫外消毒对ARB去除率仅为18.2%~40.9%,且AMP含量最高;另外,消毒技术对ERY有选择性去除效果,对其他4种ARB无明显选择性去除效果(P0.05);最后结合次氯酸钠、臭氧、紫外消毒处理对COD、NH_4~+-N的去除效果,确定强化去除ARB的消毒方式中臭氧,次氯酸钠和紫外的最佳浓度和剂量依次为5.0 mg·L~(-1)、25.0 mg·L~(-1)和45.0 m J·cm~(-2),ARB去除率依次为45.5%~74.5%、66.1%~85.5%、68.6%~85.5%,另外次氯酸钠耦合紫外消毒强化去除ARB的效果更佳.     

四环素胁迫对Shigella flexneri细菌四环素抗性基因抗性表达的影响过程

四环素(TC)抗生素在不同环境介质中已被广泛检出,为研究其对四环素抗性基因(TC-ARGs)丰度变化及表达水平的影响过程,以从活性污泥中筛选和纯化分离获得的弗氏志贺氏菌(Shigella flexneri)为研究对象,考察了不同浓度TC对其生长过程的作用影响,采用荧光定量PCR和逆转录PCR方法定量检测了不同抗性机制TC-ARGs,包括tetC、tetO和tetX基因的丰度变化及表达水平,并探讨了TC浓度与TC-ARGs丰度及其表达水平之间的相关关系.结果表明,在培养周期内(24 h),TC胁迫对Shigella flexneri细菌的生长具有抑制作用,细菌细胞浓度增长速率随TC暴露浓度的升高而降低,但对TC-ARGs丰度变化影响较小.TC胁迫能够促进Shigella flexneri细菌TC-ARGs的转录表达,tetC、tetO和tetX基因表达水平在整个培养周期内均先升高后降低.由相关性分析可知,TC浓度与TC-ARGs丰度及其表达水平之间相关关系不显著,但tetC和tetO基因丰度与其转录表达水平之间存在显著的正相关关系,表明其基因丰度一定程度上可用来衡量和评价其抗性表达水平.     

污水厂二级出水中抗生素抗性细菌的紫外灭活与复活特性

抗生素抗性细菌(ARB)作为新兴污染物受到了广泛关注,但紫外线(UV)消毒对ARB去除效果的研究尚不够充分。以某城市污水处理厂过滤后的二级出水为研究对象,通过分析UV消毒前后水中四环素、氨苄西林、氯霉素及链霉素抗性细菌丰度的变化,探究了UV消毒对ARB的去除作用及ARB的光复活与暗修复潜能。结果表明:UV对过滤后二级出水中的4种ARB具有一定灭活能力,4种ARB对UV消毒的耐受能力由高到低分别为:氨苄西林抗性细菌>四环素抗性细菌>链霉素抗性细菌≈氯霉素抗性细菌。当UV消毒剂量为20 mJ/cm2时,氯霉素与链霉素抗性细菌可被完全灭活,消毒后24 h内,这2种ARB未出现光复活或暗修复现象,这一剂量的UV不能完全灭活氨苄西林与四环素抗性细菌,且被灭活后的ARB可实现部分光复活及暗修复。当UV消毒剂量达到80 mJ/cm2时,这4种ARB均被全部灭活,消毒后6 h内,氨苄西林抗性细菌出现了复活现象。不论是在光照还是避光的条件下,UV消毒24 h后水中的总异养菌群中有超过70.32%的细菌对氨苄西林具有抗性,因此,单独UV消毒并不能有效地控制ARB从污水处理厂向环境中的传播。     

四环素抗生素对污泥中四环素抗性基因丰度和表达水平的作用影响

为研究四环素(TC)抗生素对其抗性基因(TC-ARGs)转录表达的作用影响,以从活性污泥中筛选获得的四环素抗性细菌(TRB)作为研究对象,采用荧光定量PCR和逆转录PCR方法检测了7种TC-ARGs,包括tet A、tet C、tet G、tet M、tet O、tet W和tet X基因的丰度和表达水平,并探讨了TC与TC-ARGs丰度及其表达水平之间的相关关系.结果表明,在整个培养周期内,tet A、tet G和tet W基因丰度随TC暴露浓度的增大总体呈现上升趋势,但其余TC-ARGs基因丰度整体波动较大. TC胁迫对不同TC-ARGs的转录表达水平作用影响差别较大,其中,tet A基因表达水平相对稳定,且随TC浓度的升高而上调,在TC浓度为100 mg·L^-1时,其上调倍数高达5. 3倍.在短期(1 d) TC胁迫下,TC-ARGs转录表达水平随TC浓度的升高整体呈现上调趋势.由相关性分析可知,tet A和tet W基因丰度与其转录表达水平之间存在显著的线性相关性,表明其基因丰度可在一定程度上衡量和评价其抗性表达水平,进而反映其功能活性和环境风险.     

BPAC-UF对二级出水中抗生素抗性基因去除效能研究

该文采用生物粉末活性炭(BPAC)-超滤(UF)组合工艺去除某污水厂二级出水中抗生素抗性基因(ARGs)和可溶性有机碳(DOC),并研究了ARGs在不同分子量区间的分布以及存在形态。结果表明:较直接UF工艺,BPAC-UF组合工艺对二级出水中ARGs和DOC的去除效果更优,组合工艺对tet A、tet C、tet X和sulⅠ的去除量为101.02～102.19,对DOC去除率为36.5%;组合工艺对不同类型有机物的去除效果更好,其对酪氨酸类蛋白质有机物去除率为21.7%,对其他类型有机物去除率均在60%以上;组合工艺膜出水中ARGs主要集中在大于30 kDa分子量区间,并且游离态ARGs浓度高于细胞态ARGs浓度;较直接UF,BPAC-UF组合工艺对细胞态和游离态ARGs的去除效果更好,去除率均在90%以上。     

Characterization and source analysis of indoor/outdoor culturable airborne bacteria in a municipal wastewater treatment plant

The potential health risks of airborne bacteria emission from a wastewater treatment process have been concerned. However, few studies have investigated the differences in community structure between indoor and outdoor bacteria. In this work, the characterization of airborne bacteria was studied in a municipal wastewater treatment plant in Beijing, China. Two indoor (i.e., fine screen room and sludge dewatering house) and two outdoor (i.e., aeration tank and control site) sampling sites were selected. An Andersen six-stage impactor was used for collecting culturable airborne bacteria in the air, and Illumina MiSeq sequencing was conducted to track the emission source of the culturable airborne bacteria. The results indicate that, compared with the outdoor aeration tank site, the concentrations of culturable airborne bacteria in the indoor fine screen room with poor ventilation were more than ten times higher and the particle size was about twice as large. The community structures of indoor and outdoor culturable airborne bacteria were obviously different. Enterobacteriaceae and opportunistic pathogens were detected in indoor culturable airborne bacteria, with wastewater and sludge dewatering machine identified as the primary sources. Conversely, Enterobacteriaceae and opportunistic pathogens were not detected in outdoor culturable airborne bacteria. Outdoor high wind speed might have resulted in rapid dilution and mixing of culturable airborne bacteria generated from the aeration tank with the ambient air. The results of the present research suggest that covering pollution sources, increasing ventilation rates, and using protective measures for personnel should be implemented to decrease the exposure risk to indoor culturable airborne bacteria.     

Quantification of multi-antibiotic resistant opportunistic pathogenic bacteria in bioaerosols in and around a pharmaceutical wastewater treatment plant

Pharmaceutical wastewater treatment plants(WWTPs) are thought to be a "seedbed" and reservoirs for multi-antibiotic resistant pathogenic bacteria which can be transmitted to the air environment through aeration. We quantified airborne multi-antibiotic resistance in a full-scale plant to treat antibiotics-producing wastewater by collecting bioaerosol samples from December2014 to July 2015. Gram-negative opportunistic pathogenic bacteria(GNOPB) were isolated, and antibiotic susceptibility tests against 18 commonly used antibiotics, including 11 β-lactam antibiotics, 3 aminoglycosides, 2 fluoroquinolones, 1 furan and 1 sulfonamide, were conducted.More than 45% of airborne bacteria isolated from the pharmaceutical WWTP were resistant to three or more antibiotics, and some opportunistic pathogenic strains were resistant to 16 antibiotics, whereas 45.3% and 50.3% of the strains isolated from residential community and municipal WWTP showed resistance to three or more antibiotics. The calculation of the multiple antibiotic resistance(MAR) index demonstrated that the air environment in the pharmaceutical WWTP was highly impacted by antibiotic resistance, while the residential community and municipal WWTP was less impacted by antibiotic resistance. In addition, we determined that the dominant genera of opportunistic pathogenic bacteria isolated from all bioaerosol samples were Acinetobacter, Alcaligenes, Citrobacter, Enterobacter, Escherichia, Klebsiella, Pantoea, Pseudomonas and Sphingomonas. Collectively, these results indicate the proliferations and spread of antibiotic resistance through bioaerosols in WWTP treating cephalosporin-producing wastewater, which imposed a potential health risk for the staff and residents in the neighborhood, calling for administrative measures to minimize the air-transmission hazard.     

Functional relationship between ammonia-oxidizing bacteria and ammonia-oxidizing archaea populations in the secondary treatment system of a full-scale municipal wastewater treatment plant

The abundance of ammonia-oxidizing bacteria and archaea and their amoA genes from the aerobic activated sludge tanks, recycled sludge and anaerobic digesters of a full-scale wastewater treatment plant (WWTP) was determined. Polymerase chain reaction and denaturing gradient gel electrophoresis were used to generate diversity profiles, which showed that each population had a consistent profile although the abundance of individual members varied. In the aerobic tanks, the ammonia-oxidizing bacterial (AOB) population was more than 350 times more abundant than the ammonia-oxidizing archaeal (AOA) population, however in the digesters, the AOA population was more than 10 times more abundant. Measuring the activity of the amoA gene expression of the two populations using RT-PCR also showed that the AOA amoA gene was more active in the digesters than in the activated sludge tanks. Using batch reactors and ddPCR, amoA activity could be measured and it was found that when the AOB amoA activity was inhibited in the anoxic reactors, the expression of the AOA amoA gene increased fourfold. This suggests that these two populations may have a cooperative relationship for the oxidation of ammonia.     

南昌某城市污水处理厂进水浓度低成因分析研究

针对南昌某城市污水处理厂进水浓度低的现象,为明确造成污水处理厂进水浓度低的原因,通过对南昌市2018—2020年降雨特征对污水处理厂进水污染物浓度影响的统计分析,估算了降水对雨污合流制管网污水浓度的影响.考察了合流制管网4个污水提升泵站、污水处理厂的日处理水量,结合水量平衡三角法,核算地下水、河水、雨水的渗入.以及利用倒排查法对源居民小区内/外-污水提升泵站-污水处理厂的水质沿程采样分析,得出相关结论.研究结果表明：①2018—2019年上半年污水处理厂进水COD在降雨后2~5 h进水浓度明显降低;但2019年下半年—2020年这种相关性明显削弱.②污水处理厂日进水COD受泵站的日流量的影响,呈显著负相关.③水量三角平衡定量分析表明地下水、河水混入28%~38%,雨水混入11%~18%.④水质沿程的分析显示小区到某些泵站的浓度明显降低,雨天COD、氨氮浓度分别减少了69.78%、75.21%,晴天COD、氨氮浓度分别降低了66.78%、76.94%,表明相应的小区外-泵站段的市政管网是污水处理厂进水浓度降低的关键管段,亟需进一步重点排查和改造.⑤老旧小区排水浓度受天气的影响远比新小区大,雨天分别比晴天减少79.23%、67.87%和54.40%,可能发生了大量雨水混入.     

Characterization of the airborne bacteria community at different distances from the rotating brushes in a wastewater treatment plant by 16S rRNA gene clone libraries

Biological risks of bioaerosols emitted from wastewater treatment processes have attracted wide attention in the recent years. However, the culture-based analysis method has been mostly adopted for detecting the bacterial community in bioaerosols, which may result in the underestimation of total microorganism concentration as not all microorganisms are cultivable. In this study, oligonucleotide fingerprinting of 16S rRNA genes was applied to reveal the composition and structure of the bacterial community in bioaerosols from an Orbal oxidation ditch in a Beijing wastewater treatment plant (WWTP). Bioaerosols were collected at different distances from the aerosol source, rotating brushes, and the sampling height was 1.5 m which is the common respiratory height of a human being. The bacterial communities of bioaerosols were diverse, and the lowest bacterial diversity was found at the sampling site just after the rotating brush rotating brush. A large proportion of bacteria in bioaerosols were affiliated with Proteobacteria and Bacteroidetes. Numerous bacteria present in the bioaerosols also emerged in water, indicating that the bacterial community in the bioaerosols was related to that of the aerosols’ sources. The forced aeration of rotating brushes brought about observably distinct bacterial communities between sampling sites situated before and after the rotating brush. Isolation sources of closest relatives in bioaerosols clone libraries were associated with the aqueous environment in the WWTP. Common potential pathogens in bioaerosols as well as those not reported in previous research were also analyzed in this study. Measures should be adopted to reduce the emission of bioaerosols and prevent their exposure to workers.     

Optimum municipal wastewater treatment plant design with consideration of uncertainty

A newly developed model for the optimum municipal wastewater treatment plant (MWTP) design is presented. Through introducing the interval variables, the model attempts to consider the effects of uncertainties caused by the fluctuation of the wastewater quality and quantity during the design of MWTP. The model solution procedure is illustrated in detail, and a numerical example is given to verify the feasibility and advantage of the model. Furthermore, the possibility of the model application is briefly outlined.     

Monitoring of 943 organic micropollutants in wastewater from municipal wastewater treatment plants with secondary and advanced treatment processes

To perform a systematic survey on the occurrence and removal of micropollutants during municipal wastewater treatment, 943 semi-volatile organic chemicals in 32 wastewater samples including influents of secondary treatments, secondary effluents and final effluents(effluents of advanced treatments), which were collected from seven full-scale municipal wastewater treatment plants(MWTPs) in China, were examined by gas chromatography-mass spectrometry(GC-MS) coupled with an automated identification and quantification system with a database(AIQS-DB). In total, 196 and 145 chemicals were detected in secondary and final effluents, respectively. The majority of the total concentrations(average removal efficiency, 87.0%±5.9%) of the micropollutants were removed during secondary treatments. However, advanced treatments achieved different micropollutant removal extents from secondary effluents depending on the different treatment processes employed. Highly variable removal efficiencies of total concentrations(32.7%–99.3%) were observed among the different advanced processes. Among them,ozonation-based processes could remove 70.0%–80.9% of the total concentrations of studied micropollutants. The potentially harmful micropollutants, based on their detection frequency and concentration in secondary and final effluents, were polycyclic aromatic hydrocarbons(PAHs)(2-methylnaphthalene, fluoranthene, pyrene, naphthalene and phenanthrene), phosphorus flame retardants(tributyl phosphate(TBP), tris(2-chloroethyl)phosphate(TCEP) and tris(1,3-dichloro-2-propyl) phosphate(TDCP)), phthalates(bis(2-ethylhexyl)phthalate(DEHP)), benzothiazoles(benzothiazole,2-(methylthio)-benzothiazol, and 2(3H)-benzothiazolone) and phenol. This study indicated that the presence of considerable amounts of micropollutants in secondary effluent creates the need for suitable advanced treatment before their reuse.