Inactivation, reactivation and regrowth of indigenous bacteria in reclaimed water after chlorine disinfection of a municipal wastewater treatment plant

Disinfection of reclaimed water prior to reuse is important to prevent the transmission of pathogens. Chlorine is a widely utilized disinfectant and as such is a leading contender for disinfection of reclaimed water. To understand the risks of chlorination resulting from the potential selection of pathogenic bacteria, the inactivation, reactivation and regrowth rates of indigenous bacteria were investigated in reclaimed water after chlorine disinfection. Inactivation of total coliforms, Enterococcus and Salmonella showed linear correlations, with constants of 0.1384, 0.1624 and 0.057 L/(mg·min) and R² of 0.7617, 0.8316 and 0.845, respectively. However, inactivation of total viable cells by measurement of metabolic activity typically showed a linear correlation at lower chlorine dose (0-22 (mg·min)/L), and a trailing region with chlorine dose increasing from 22 to 69 (mg·min)/L. Reactivation and regrowth of bacteria were most likely to occur after exposure to lower chlorine doses, and extents of reactivation decreased gradually with increasing chlorine dose. In contrast to total coliforms and Enterococcus, Salmonella had a high level of regrowth and reactivation, and still had 2% regrowth even after chlorination of 69 (mg·min)/L and 24 hr storage. The bacterial compositions were also significantly altered by chlorination and storage of reclaimed water, and the ratio of Salmonella was significantly increased from 0.001% to 0.045% after chlorination of 69 (mg·min)/L and 24 hr storage. These trends indicated that chlorination contributes to the selection of chlorine-resistant pathogenic bacteria, and regrowth of pathogenic bacteria after chlorination in reclaimed water with a long retention time could threaten public health security during wastewater reuse.     

Effects of O3/Cl2 disinfection on corrosion and opportunistic pathogens growth in drinking water distribution systems

The effects of O_3/Cl_2 disinfection on corrosion and the growth of opportunistic pathogens in drinking water distribution systems were studied using annular reactors(ARs).The corrosion process and most probable number(MPN) analysis indicated that the higher content of iron-oxidizing bacteria and iron-reducing bacteria in biofilms of the AR treated with O_3/Cl_2 induced higher Fe_3O_4 formation in corrosion scales.These corrosion scales became more stable than the ones that formed in the AR treated with Cl_2 alone.O_3/Cl_2 disinfection inhibited corrosion and iron release efficiently by changing the content of corrosion-related bacteria.Moreover,ozone disinfection inactivated or damaged the opportunistic pathogens due to its strong oxidizing properties.The damaged bacteria resulting from initial ozone treatment were inactivated by the subsequent chlorine disinfection.Compared with the AR treated with Cl_2 alone,the opportunistic pathogens M.avium and L.pneumophila were not detectable in effluents of the AR treated with O_3/Cl_2,and decreased to(4.60 ± 0.14) and(3.09 ± 0.12) log10(gene copies/g corrosion scales) in biofilms,respectively.The amoeba counts were also lower in the AR treated with O_3/Cl_2.Therefore,O_3/C_l2 disinfection can effectively control opportunistic pathogens in effluents and biofilms of an AR used as a model for a drinking water distribution system.     

Full-scale study of removal effect on Cyclops of zooplankton with chlorine dioxide

Cyclops of zooplankton propagated excessively in eutrophic water body and could not be effectively inactivated by the conventional disinfections process like chlorination due to its stronger resistance to oxidation. In this paper, a full-scale study of chlorine dioxide preoxidation cooperating with routine clarification process for Cyclops removal was conducted in a waterworks. The experimental results were compared with that of the existing prechlorination process in several aspects: including the Cyclops removal efficiencies of water samples taken from the outlets of sedimentation tank and sand filter and the security of drinking water and so on. The results showed that chlorine dioxide might be more effective to inactivate Cyclops than chlorine and Cyclops could be thoroughly removed from water by pre-dosing chlorine dioxide process. The GC-MS examination and Ames test further showed that the sort and amount of organic substance in the treated water by chlorine dioxide preoxidation were evidently less than that of prechlorination and the mutagenicity of drinking water treated by pre-dosing chlorine dioxide was substantially reduced compared with prechlorination.     

Transformation of sulfamethazine during the chlorination disinfection process: Transformation, kinetics, and toxicology assessment

Various disinfection byproducts(DBPs) form during the process of chlorination disinfection,posing potential threats to drinking water safety and human health. Sulfamethazine(SMT),the most commonly used and frequently detected veterinary antibiotic, was investigated in detail with regard to its transformation and kinetics in reactions with free available chlorine(FAC). Using liquid chromatography coupled to quadrupole time-of-flight tandem mass spectrometry, several DBPs were identified based on different confidence levels, and a variety of reaction types, including desulfonation, S–N cleavage, hydroxylation, and chlorine substitution, were proposed. The kinetic experiments indicated that the reaction rate was FAC-and pH-dependent, and SMT exhibits low reactivity toward FAC in alkaline conditions. The DBPs exhibited a much higher acute toxicity than SMT, as estimated by quantitative structure activity relationship models. More importantly, we observed that the FAC-treated SMT reaction solution might increase the genotoxic potential due to the generation of DBPs. This investigation provides substantial new details related to the transformation of SMT in the chlorination disinfection process.     

Total organic halogen (TOX) in human urine: A halogen-specific method for human exposure studies

Disinfection by-products(DBPs) are a complex mixture of compounds unintentionally formed as a result of disinfection processes used to treat drinking water. Effects of long-term exposure to DBPs are mostly unknown and were the subject of recent epidemiological studies. However,most bioanalytical methods focus on a select few DBPs. In this study, a new comprehensive bioanalytical method has been developed that can quantify mixtures of organic halogenated compounds, including DBPs, in human urine as total organic chlorine(TOCl), total organic bromine(TOBr), and total organic iodine(TOI). The optimized method consists of urine dilution, adsorption to activated carbon, pyrolysis of activated carbon, absorption of gases in an aqueous solution, and halide analysis with ion chromatography and inductively coupled plasma-mass spectrometry. Spike recoveries for TOCl, TOBr, and TOI measurements ranged between 78% and 99%. Average TOCl, TOBr, and TOI concentrations in five urine samples from volunteers who consumed tap water were 1850, 82, and 21.0 μg/L as X~-, respectively.Volunteers who consumed spring water(control) had TOCl, TOBr, and TOI average concentrations in urine of 1090, 88, and 10.3 μg/L as X~-, respectively. TOCl and TOI in the urine samples from tap water consumers were higher than the control. However, TOBr was slightly lower in tap water urine samples compared to mineral water urine samples, indicating other sources of environmental exposure other than drinking water. A larger sample population that consumes tap water from different cities and mineral water is needed to determine TOCl, TOBr, and TOI exposure from drinking water.     

Inactivation effect of pressurized carbon dioxide on bacteriophage Qβ and ΦX174 as a novel disinfectant for water treatment

The inactivation effects of pressurized CO2 against bacteriophage Qβ and ΦX174 were investigated under the pressure of 0.3–0.9 MPa, initial concentration of 107–109PFU/mL, and temperature of17.8°C–27.2°C. The optimum conditions were found to be 0.7 MPa and an exposure time of 25 min. Under identical treatment conditions, a greater than 3.3-log reduction in bacteriophage Qβ was achieved by CO2, while a nearly 3.0 log reduction was observed for phage ΦX174. The viricidal effects of N2O(an inactivation gas with similar characteristics to CO2), normal acid(HCl), and CO2 treatment with phosphate buffered saline affirmed the chemical nature of CO2 treatment. The pumping cycle, depressurization rate, and release of intracellular substances caused by CO2 were its viricidal mechanisms. The results indicate that CO2 has the potential for use as a disinfectant without forming disinfection by-products.     

Assessment and management of the performance risk of a pilot reclaimed water disinfection process

Chlorination disinfection has been widely used in reclaimed water treatment plants to ensure water quality. In order to assess the downstream quality risk of a running reclaimed water disinfection process, a set of dynamic equations was developed to simulate reactions in the disinfection process concerning variables of bacteria, chemical oxygen demand （COD）, ammonia and monochloramine. The model was calibrated by the observations obtained from a pilot disinfection process which was designed to simulate the actual process in a reclaimed water treatment plant. A Monte Carlo algorithm was applied to calculate the predictive effluent quality distributions that were used in the established hierarchical assessment system for the downstream quality risk, and the key factors affecting the downstream quality risk were defined using the Regional Sensitivity Analysis method. The results showed that the seasonal upstream quality variation caused considerable downstream quality risk; the effluent ammonia was significantly influenced by its upstream concentration; the upstream COD was a key factor determining the process effluent risk of bacterial, COD and residual disinfectant indexes; and lower COD and ammonia concentrations in the infiuent would mean better downstream quality.     

Intensified inactivation of model and environmental bacteria by an atmospheric-pressure air-liquid discharge plasma compared with chlorination

Water-borne pathogenic bacteria are always the top priority to be removed through disinfection process in water treatment due to their threat to human health. It was necessary to develop novel disinfection methods since the conventional chlorine disinfection was inefficient in inactivating chlorine-resistant bacteria, inducing the viable but non-culturable (VBNC) bacteria and forming disinfection by-products (DBPs). In this study, the inactivation of four model strains including Gram-negative (G⁻), Gram-positive (G⁺) and environmental samples by atmospheric-pressure air-liquid discharge plasma (ALDP) was assessed systematically. The results showed that ALDP was superior in inactivating all of the samples compared with chlorination. During 10 min ALDP treatment, the G⁻ bacteria were completely inactivated, and the G⁺ one was inactivated by more than 4.61 logs. The inactivation of bacteria from a campus lake and a wastewater treatment plant effluent exceeded 99.82% and 97.78%, respectively. For G⁻ bacteria, ALDP resulted in a much lower (10²∼10³ times) levels of VBNC cells than chlorination. ALDP could effectively remove the chlorine-resistant bacteria. More than 96.41% of the intracellular DNA and 99.99% of the extracellular DNA were removed, whereas it was only 56.35% and 12.82% for chlorination. ALDP had a stronger ability to destroy cell structure than chlorination, presumably due to the existence of ROS (·OH, ¹O₂ and O₂⁻). GC-MS analysis showed that ALDP produced less DBPs than chlorination. These findings provided new insights for the application of discharge plasma in water disinfection, which could be complemental or alternative to the conventional disinfection methods.     

Reduction behaviors of permanganate by microbial cells and concomitant accumulation of divalent cations of Mg2+, Zn2+, and Co2+

Permanganate treatment is widely used for disinfection of bacteria in surface-contaminated water. In this paper, the fate of the dissolved permanganate in aqueous solution after contact with cells of Pseudomonas fluorescens was studied. Concomitant accumulation of divalent cations of Mg~(2+), Zn~(2+), and Co~(2+) during precipitation of Mn oxides was also studied. The time course of the Mn concentration in solution showed an abrupt decrease after contact of Mn(VII) with microbial cells, followed by an increase after ～ 24 hr.XRD analysis of the precipitated Mn oxides, called biomass Mn oxides, showed the formation of low-crystalline birnessite. Visible spectroscopy and X-ray absorption near edge structure(XANES) analyses indicated that dissolved Mn(VII) was reduced to form biomass Mn oxides involving Mn(IV) and Mn(III), followed by reduction to soluble Mn(II).The numbers of electron transferred from microbial cells to permanganate and to biomass Mn oxides for 24 hr after the contact indicated that the numbers of electron transfer from microbial cell was approximately 50 times higher to dissolved permanganate than to the biomass Mn oxides in present experimental conditions. The 24 hr accumulation of divalent cations during formation of biomass Mn oxides was in the order of Co~(2+) Zn~(2+) Mg~(2+).XANES analysis of Co showed that oxidation of Co~(2+) to Co~(3+) resulted in higher accumulation of Co than Zn and Mg. Thus, treatment of surface water by KMnO_4 solution is effective not only for disinfection of microorganisms, but also for the elimination of metal cations from surface water.     

Investigation of UV-TiO2 photocatalysis and its mechanism in Bacillus subtilis spore inactivation

The inactivation levels of Bacillus subtilis spores for various disinfection processes （ultraviolet （UV）, TiO2 and UV-TiO2） were compared. The results showed that the inactivation effect of B. subtilis spores by UV treatment alone was far below that for bacteria without endospores. TiO2 alone in the dark, as a control experiment, exhibited almost no inactivation effect. Compared with UV treatment alone, the inactivation effect increased significantly with the addition of TiO2. Increases of the UV irradiance and Ti02 concentration both contributed to the increase of the inactivation effect. Lipid peroxidation was found to be the underlying mechanism of inactivation. Malondialdehyde （MDA）, the degradation product of lipid peroxidation, was used as an index to determine the extent of the reaction. The MDA concentration surged surprisingly to 3.24 nmol/mg dry cell with the combination disinfection for 600 see （0.10 mW/cm2 irradiance and 10 mg/L TiO2）. In contrast, for UV alone or TiO2 in the dark, the MDA concentration was 0.38 and 0.25 nmol/mg dry cell, respectively, under the same conditions. This indicated that both UV and TiO2 were essential for lipid peroxidation. Changes in cell ultrastructure were observed by transmission electron microscopy. The cell membrane was heavily damaged and cellular contents were completely lysed with the UV-TiO2 process, suggesting that lipid peroxidation was the root of the enhancement in inactivation efficiency.     

二氧化氯对不同微生物的灭活特性及其对群落结构特征的影响

二氧化氯是一种性能优良、应用广泛的消毒剂,可通过破坏细胞或病毒的组成结构、阻碍细胞代谢等方式实现微生物灭活。在自配水条件下,以二氧化氯投加量×消毒时间计算,二氧化氯剂量在15（mg·min）/L时,可实现对常见病毒（包括肠病毒71型、大肠杆菌噬菌体MS2等）3 log以上灭活率,在60（mg·min）/L时,可实现对常见细菌（包括大肠杆菌、金黄色葡萄球菌等）1.5 log以上灭活率,但灭活隐孢子虫卵则需要更高的剂量（如1.9 log灭活率可能需约600（mg·min）/L剂量）;在实际污水厂进水中,30（mg·min）/L二氧化氯剂量只能分别实现0.8 log和0.5 log的大肠杆菌和总大肠菌群灭活率。二氧化氯消毒效果随温度升高显著提升,对于不同微生物,pH的变化对二氧化氯消毒效果的影响可能存在不同,而水中的有机物通常会因消耗二氧化氯而降低消毒效果,但在自然水体中也存在由于天然有机物可能的影响导致消毒效果优于自配水的情况。关于二氧化氯消毒后细菌群落结构的变化研究不多,仅有少量研究涉及市政污水、再生水、饮用水等。二氧化氯消毒一定时间后,悬浮态和生物膜上的微生物均可能出现再生长现象,但再生长过程中这些残生细菌的群落结构变化及其生长分泌特性仍有待研究。     

氯消毒对产ESBLs菌β-内酰胺酶类抗性基因接合转移的抑制作用

以产超广谱β-内酰胺酶（ESBLs）的大肠杆菌T413和NK5449分别作为供体菌和受体菌进行接合试验,研究氯消毒处理供体菌对β-内酰胺酶类抗性基因接合转移的抑制作用.结果表明,供体菌质粒上的bla_CTX-M和bla_TEM基因可接合转移至受体菌,接合转移频率为6.57×10^-2.当氯消毒接触时间为30min,有效氯浓度在0.25~1.5mg/L时,接合转移频率未出现数量级的降低,而有效氯浓度为2mg/L时,接合转移频率则骤降至2.02×10^-5.在CT值为60（mg·min）/L的各种组合中,4mg/L×15min对接合转移的抑制效果最佳.氯消毒后的供体菌数量与接合转移频率呈正相关.经高剂量氯消毒后的供体菌具有较高的再生长率,但接合转移频率却很低.高剂量氯消毒会损伤供体菌菌体结构和功能,使胞外分泌物减少,阻碍质粒传递.     

“红虫”杀灭前后自来水的安全评价

采用发光细菌急性毒性试验,配合大型蚤和斑马鱼胚胎毒性试验验证,测定有"红虫"存在的自来水以及用氯胺、二氧化氯和过氧化氢对"红虫"进行杀灭后自来水的生物毒性,判定其对人体健康可能造成的危害。结果显示:"红虫"自来水并没有检测出明显的生物毒性;氯胺在起始剂量不超过4mg/L时不仅有较好的控制效果,而且用其杀灭"红虫"后的自来水具有较低的健康风险;采用低浓度过氧化氢(0.005%)对"红虫"进行杀灭,其杀灭后自来水仍具有较强生物毒性,最终认为不宜作为自来水常规消毒剂;使用6～8mg/L的二氧化氯可有效杀灭"红虫",杀灭后自来水的生物毒性为低毒,其最终消毒副产物对人体的健康风险较低。     

不同消毒方式对饮用水生物活性炭出水消毒效果的对比研究

针对水厂生物活性炭深度处理工艺可能产生的细菌泄露问题,采用上海徐泾水厂生物活性炭出水,通过对异养菌平板计数和卤代烃气相色谱检测分析,比较了次氯酸钠和氯胺2种消毒方式对出厂水水质安全保障的效果.在水温30℃下,采用NaClO消毒剂进行消毒处理时,初始余氯值达到1.84 mg/L、接触30 min即可保障灭活率lg(N0/...     

给水管网中耐氯性细菌的灭活特性研究

使用4种常见消毒剂对从实际管网中分离出来的7株耐氯性细菌进行消毒实验.结果表明,这7株细菌均具有较高的耐氯性,其中1株耐氯性最高的类龟分支杆菌自由氯99.9%灭活的CT值为120 mg.(L.min)-1,另外2株血红鞘氨醇单胞菌和甲基杆菌99.9%灭活的CT值分别为7 mg.(L.min)-1和4 mg.(L.min)-1.比较4种消毒剂的消毒效果发现,二氧化氯和单过硫酸氢钾的消毒效果较好,能够在30 min内使分支杆菌的灭活率达到5个数量级.自由氯由于衰减较快,消毒效果不佳.一氯胺能够维持一定的消毒剂浓度,但由于其氧化性较弱,因此需要提高浓度,才能满足消毒要求.能在1 h内灭活3个数量级以上分支杆菌的消毒剂投加量为:3.0 mg/L一氯胺、1.0 mg/L二氧化氯(以Cl2计)和1.0 mg/L单过硫酸氢钾(以Cl2计).考虑到我国水厂消毒的实际情况,建议采用间歇性提高一氯胺浓度或改换二氧化氯消毒的方法,提高对耐氯性细菌的灭活效果.     

给水管网管壁铁细菌生长特性模拟及控制对策研究

利用AR反应器对管壁铁细菌的生长特性、影响因素以及悬浮铁细菌与管壁铁细菌的相关关系进行了研究.管壁铁细菌在反应器运行12 d后达到稳定生长,出水中铁细菌数量比进水增加1 lg,剪切力对于铁细菌达到稳定生长的时间有影响,但对稳定生长后挂片上的铁细菌数量没有明显的影响.稳定余氯0.3 mg/L可以有效控制悬浮和管壁铁细菌的生长,数量降低1 lg;对于管垢内的铁细菌即使余氯增加到1.0 mg/L仍无法杀灭;余氯浓度0.05 mg/L无法有效控制悬浮和管壁附着铁细菌的生长.对于稳定生长下的管壁铁细菌,增加余氯浓度至1.25 mg/L,可使管壁铁细菌数量降低2 lg~3 lg.稳定生长状态下,悬浮铁细菌与管壁铁细菌数量具有线性关系.并在试验基础上提出给水管网铁细菌生长控制对策:维持管网余氯0.3 mg/L;定期采用较高浓度消毒剂冲洗管道;采用新型防腐管材或加快旧管网改造,提高管网抗腐蚀性能等.     

二氧化氯对水中流感病毒的消毒效果

通过考察MDCK细胞病变程度,研究二氧化氯(ClO₂)对水中流感病毒(包括流感病毒Ⅰ型 、流感病毒Ⅱ型和流感病毒Ⅲ型)的消毒效果.对各种试验条件,如消毒剂投加量、消毒时间 及pH值等影响因素进行了研究,并与液氯(Cl₂)消毒效果进行了对比.结果表明,ClO₂投加量为 40mg/L时,作用20min,ClO₂对试验的流感病毒具有很好的灭活效果,且在pH值为3.0～8.0的范 围内均达到很好的消毒效果.而液氯在试验的投加量、时间和pH值范围内均不能灭活流感病 毒.      

再生水紫外线-氯联合消毒工艺特性研究

以二级出水作为消毒对象,比较了紫外线-氯联合消毒与单一紫外线消毒对水中肠道指示菌的灭活效果,考察了联合消毒工艺副产物(三卤甲烷)生成情况.结果表明,单一紫外线消毒对肠道指示菌的灭活曲线在剂量大于10mJ/cm2时存在拖尾现象,剂量为80mJ/cm2时对总大肠菌群最高对数灭活率约为2.5;而在紫外线剂量20mJ/cm2、氯投加量8mg/L(接触时间30min)条件下,紫外线-氯联合消毒对总大肠菌群的对数灭活率达到7.0.20mJ/cm2紫外线与8mg/L氯消毒组合及80mJ/cm2紫外线与3mg/L氯消毒组合均可有效灭活总大肠菌群浓度至3CFU/L以下.经紫外线照射后再氯消毒,二级出水中三卤甲烷生成量仅为10~55mg/L.80mJ/cm2紫外线照射再氯消毒后三卤甲烷生成量略高于20mJ/cm2紫外线照射再氯消毒的情形.紫外线消毒后投加氯消毒,可有效提高再生水消毒效果,控制消毒副产物生成量.     

氯消毒和紫外消毒对城市污水处理厂二沉池出水中粪大肠菌群耐药性的影响

为考察城市污水处理厂消毒方式对出水中细菌耐药性的影响,使用二沉池出水进行氯消毒和紫外消毒试验. 选择AMP(氨苄西林)、TET(盐酸四环素)、CIP(环丙沙星)和CHL(氯霉素)作为典型的抗生素,研究消毒前后粪大肠菌群耐药率的变化. 结果表明:当水中ρ(有效氯)在0~1.0mg/L范围内增加时,粪大肠菌群对AMP和CIP的耐药率都有所升高,而对TET的耐药率却呈下降趋势. 当使用1.0mg/L氯消毒之后,粪大肠菌群对AMP、TET、CIP、CHL的耐药率分别为35.1%、5.6%、62.3%、0. 粪大肠菌群对AMP、TET和CIP的耐药性在氯消毒后会逐渐恢复. 氯消毒后48h,粪大肠菌群对AMP、TET和CIP的耐药率均高于消毒前. 在较低的紫外辐照剂量 (约16mJ/cm2以下)范围内,随着紫外辐照剂量的升高,粪大肠菌群对多种抗生素的耐药率均有降低,而较高的紫外辐照剂量则可能导致粪大肠菌群耐药率升高.