Evaluation of laboratory-made sludge for an anaerobic biodegradability test and its use for assessment of 13 chemicals

Laboratory-made sludge for a biogas based anaerobic biodegradability test was prepared as an alternative for digested sludge from wastewater treatment plants (WWTPs). Biodegradation activities and background gas productions of digested sludge from various WWTPs were found to vary significantly depending on the source, which adversely affected test reliability. Subsequently, test conditions such as sludge concentration and sludge washing were examined with the laboratory-made sludge and a sludge concentration of 1.0 g-SS/L without washing was determined to be most suitable. Under these conditions, biodegradability tests were conducted for 13 select chemicals and their relative toxicities to methanogenic bacteria were evaluated. The results of biodegradability tests showed that chemicals with -OH and -CH2OH radicals were readily biodegraded and those with -Cl, -NO2, -NH2, -SO3H and -CH3 had inhibited degradation responses m-nitriphenol and 2,4,6-trichlorophenol were highly toxic to methanogenic bacteria, with m-nitrophenol completely inhibiting methane fermentation as low as 20 mg/L.     

不同污泥源厌氧氨氧化污泥的比较

采用厌氧序批式反应器,以好氧硝化污泥和厌氧颗粒污泥为污泥源,通过对氨氮、亚硝酸盐氮、pH等指标的监测和数据分析、污泥颜色变化和菌落电镜照片的观察,研究2种不同污泥源厌氧氨氧化污泥的差异.结果表明,不同污泥源厌氧氨氧化污泥的颜色不同,污泥中具有厌氧氨氧化作用的优势菌不同;由厌氧颗粒污泥为污泥源培养出的厌氧氨氧化污泥具有较高的厌氧氨氧化活性.     

接种不同污泥源条件下厌氧氨氧化菌的特性

2组ASBR接种污泥源分别为好氧硝化污泥、好氧硝化污泥和厌氧氨氧化污泥按2：1比例}昆合的混合污泥。在相同条件下,经过驯化培养均实现了厌氧氨氧化的稳定运行。接种好氧硝化污泥的反应器的适应期为29d,经过105d的培养反应器成功启动;接种混合污泥的反应器的适应期为13d,经过49d反应器启动成功。从2组ASBR污泥中提取细菌总DNA,经过厌氧氨氧化菌特异引物Pla46rc／Amx820对污泥样品进行PCR扩增、克隆和测序等分析。实验结果表明,接种不同污泥源条件下的反应器中厌氧氨氧化菌的特性存在差异,接种污泥源为好氧硝化污泥的反应器中存在的厌氧氨氧化菌种为CandidatusKuenenia,而接种混合污泥的反应器中存在的厌氧氨氧化菌种为CandidatusAnammox—oglobus,与最初接种的混合污泥中的厌氧氨氧化菌相同。当接种污泥中存在厌氧氨氧化菌时,该菌株经过长时间的驯化可成为优势菌种,而当接种污泥中无厌氧氨氧化菌存在时,CandidatusKueneniasp．可以在反应器中占主导,具有更强的竞争优势。     

Isolation, identification and characterization of Bacillus amyloliquefaciens BZ-6, a bacterial isolate for enhancing oil recovery from oily sludge

Over 100 biosurfactant-producing microorganisms were isolated from oily sludge and petroleum-contaminated soil from Shengli oil field in north China. Sixteen of the bacterial isolates produced biosurfactants and reduced the surface tension of the growth medium from 71 to <30 mN m⁻¹ after 72 h of growth. These bacteria were used to treat oily sludge and the recovery efficiencies of oil from oily sludge were determined. The oil recovery efficiencies of different isolates ranged from 39% to 88%. Bacterial isolate BZ-6 was found to be the most efficient strain and the three phases (oil, water and sediment) were separated automatically after the sludge was treated with the culture medium of BZ-6. Based on morphological, physiological characteristics and molecular identification, isolate BZ-6 was identified as Bacillus amyloliquefaciens. The biosurfactant produced by isolate BZ-6 was purified and analyzed by high performance liquid chromatography-electrospray ionization tandem mass spectrometry. There were four ion peaks representing four different fengycin A homologues.     

厌氧序批式反应器中丝状颗粒污泥的形成

以葡萄糖为基质,研究浮动盖式ASBR中颗粒污泥的形成。实验结果表明,污泥在120 d时完全颗粒化,颗粒污泥具有良好的沉降性能(25 m/h)及良好的产甲烷活性,甲酸、乙酸、丙酸及丁酸最大代谢活性分别达到0.216、0.825、0.237和0.796 g COD/(g VSS.d)。电镜扫描(SEM)发现,整个污泥结构均一,均由丝状菌构成。     

城市垃圾焚烧渗沥液中Ca2+对厌氧EGSB处理效果的影响简

研究了城市生活垃圾焚烧厂渗沥液中Ca²⁺对厌氧颗粒污泥膨胀床反应器（EGSB）处理效果的影响,并采用静态实验方法考察了Ca²⁺对厌氧颗粒污泥产甲烷活性的影响。实验结果表明,进水COD为17 000 mg/L的条件下,当Ca²⁺浓度低于6 000 mg/L时,EGSB对COD去除率达93%以上;当Ca²⁺浓度高于6 000 mg/L时,COD去除率随运行时间明显下降,并在污泥中形成大量沉淀。静态实验结果表明,废水中低浓度Ca²⁺促进了厌氧颗粒污泥的产甲烷活性,但高浓度Ca²⁺明显抑制了其产甲烷活性,这是导致高Ca²⁺浓度条件下EGSB对COD去除率降低的主要原因。研究表明,颗粒污泥产甲烷活性恢复程度随Ca²⁺浓度增加而减弱。     

Isolation and characterization of mesotrione-degrading Bacillus sp. from soil

Dissipation kinetics of mesotrione, a new triketone herbicide, sprayed on soil from Limagne (Puy-de-Dôme, France) showed that the soil microflora were able to biotransform it.Bacteria from this soil were cultured in mineral salt solution supplemented with mesotrione as sole source of carbon for the isolation of mesotrione-degrading bacteria. The bacterial community structure of the enrichment cultures was analyzed by temporal temperature gradient gel electrophoresis (TTGE). The TTGE fingerprints revealed that mesotrione had an impact on bacterial community structure only at its highest concentrations and showed mesotrione-sensitive and mesotrione-adapted strains. Two adapted strains, identified as Bacillus sp. and Arthrobacter sp., were isolated by colony hybridization methods.Biodegradation assays showed that only the Bacillus sp. strain was able to completely and rapidly biotransform mesotrione. Among several metabolites formed, 2-amino-4-methylsulfonylbenzoic acid (AMBA) accumulated in the medium. Although sulcotrione has a chemical structure closely resembling that of mesotrione, the isolates were unable to degrade it.     

Long-term Hg pollution-induced structural shifts of bacterial community in the terrestrial isopod (Porcellio scaber) gut

In previous studies we detected lower species richness and lower Hg sensitivity of the bacteria present in egested guts of Porcellio scaber (Crustacea, Isopoda) from chronically Hg polluted than from unpolluted environment. Basis for such results were further investigated by sequencing of 16S rRNA genes of mercury-resistant (Hg^r) isolates and clone libraries. We observed up to 385 times higher numbers of Hg^r bacteria in guts of animals from polluted than from unpolluted environment. The majority of Hg^r strains contained merA genes. Sequencing of 16S rRNA clones from egested guts of animals from Hg-polluted environments showed elevated number of bacteria from Pseudomonas, Listeria and Bacteroidetes relatives groups. In animals from pristine environment number of bacteria from Achromobacter relatives, Alcaligenes, Paracoccus, Ochrobactrum relatives, Rhizobium/Agrobacterium, Bacillus and Microbacterium groups were elevated. Such bacterial community shifts in guts of animals from Hg-polluted environment could significantly contribute to P. scaber Hg tolerance.     

Fate of bisphenol A during treatment with the litter-decomposing fungi Stropharia rugosoannulata and Stropharia coronilla

Bisphenol A is an endocrine disrupting compound, which is ubiquitous in the environment due to its wide use in plastic and resin production. Seven day old cultures of the litter-decomposing fungus Stropharia coronilla removed the estrogenic activity of bisphenol A (BPA) rapidly and enduringly. Treatment of BPA with purified neutral manganese peroxidase (MnP) from this fungus also resulted in 100% reduction of estrogenic activity, as analyzed using a bioluminescent yeast assay, and in the formation of polymeric compounds. In cultures of Stropharia rugosoannulata, estrogenic activity also quickly disappeared but temporarily re-emerged in the further course of cultivation. LC-MS analysis of the extracted estrogenic culture liquid revealed [M−H]⁻ ions with m/z values of 219 and 235. We hypothesize that these compounds are ring fission products of BPA, which still exhibit one intact hydroxyphenyl group to interact with estrogen receptors displayed by the yeast.     

The exposition of a calcareous Mediterranean soil to toxic concentrations of Cr, Cd and Pb produces changes in the microbiota mainly related to differential metal bioavailability

The involvement of the bacterial community of an agricultural Mediterranean calcareous soil in relation to several heavy metals has been studied in microcosms under controlled laboratory conditions. Soil samples were artificially polluted with Cr(VI), Cd(II) and Pb(II) at concentrations ranging from 0.1 to 5000 mg kg⁻¹ and incubated along 28 d. The lowest concentrations with significant effects in soil respirometry were 10 mg kg⁻¹ Cr and 1000 mg kg⁻¹ Cd and Pb. However, only treatments showing more than 40% inhibition of respirometric activity led to significant changes in bacterial composition, as indicated by PCR-DGGE analyses. Presumable Cr- and Cd-resistant bacteria were detected in polluted microcosms, but development of the microbiota was severely impaired at the highest amendments of both metals. Results also showed that bioavailability is an important factor determining the impact of the heavy metals assayed, and even an inverted potential toxicity ranking could be achieved if their soluble fraction is considered instead of the total concentration. Moreover, multiresistant bacteria were isolated from Cr-polluted soil microcosms, some of them showing the capacity to reduce Cr(VI) concentrations between 26% and 84% of the initial value. Potentially useful strains for bioremediation were related to Arthrobacter crystallopoietes, Stenotrophomonas maltophilia and several species of Bacillus.     

Tissue S/N ratios and stable isotopes (δS and δN) of epilithic mosses (Haplocladium microphyllum) for showing air pollution in urban cities in Southern China

In urban cities in Southern China, the tissue S/N ratios of epilithic mosses (Haplocladium microphyllum), varied widely from 0.11 to 0.19, are strongly related to some atmospheric chemical parameters (e.g. rainwater SO₄²⁻/NH₄⁺ ratios, each people SO₂ emission). If tissue S/N ratios in the healthy moss species tend to maintain a constant ratio of 0.15 in unpolluted area, our study cities can be divided into two classes: class I (S/N > 0.15, S excess) and class II (S/N < 0.15, N excess), possibly indicative of stronger industrial activity and higher density of population, respectively. Mosses in all these cities obtained S and N from rainwater at a similar ratio. Sulphur and N isotope ratios in mosses are found significantly linearly correlated with local coal δ³⁴S and NH₄⁺-N wet deposition, respectively, indicating that local coal and animal NH₃ are the major atmospheric S and N sources.     

Isolation and characterization of lead-tolerant Ochrobactrum intermedium and its role in enhancing lead accumulation by Eucalyptus camaldulensis

In this study, the potential of rhizospheric bacteria in promoting the growth and Pb accumulation by the woody plant Eucalyptus camaldulensis under hydroponic conditions was investigated for the first time. Three Pb-tolerant bacteria were isolated from the rhizosphere of E. camaldulensis grown in Pb-contaminated soils in the Bo Ngam Pb mine, Thailand. Based on analysis of partial 16S rRNA gene sequence, the three isolates were identified as Microbacterium paraoxydans BN-2, Ochrobactrum intermedium BN-3, and Bacillus fusiformis BN-4. Among these strains, O. intermedium BN-3 showed the highest tolerance to not only Pb but also Cd and Zn. After growth in the presence of Pb, the membranes of O. intermedium BN-3 cells exhibited an increase in unsaturated fatty acid levels but a decrease in fluidity. In hydroponic studies, inoculation of O. intermedium BN-3 significantly increased the biomass and Pb accumulation by E. camaldulensis compared to the uninoculated control. The results suggested the role of the natural rhizospheric bacteria localized to the root surface of E. camaldulensis in promoting Pb accumulation and plant growth. Our results indicate that O. intermedium BN-3 and other indigenous rhizospheric bacteria have the potential to improve the efficiency of phytoremediation of Pb-contaminated sites.     

Influence of extracellular polymeric substances (EPS) on Cd adsorption by bacteria

The role of extracellular polymeric substances (EPS) in Cd adsorption by Bacillus subtilis and Pseudomonas putida was investigated using a combination of batch adsorption experiments, potentiometric titrations, Fourier transform infrared spectroscopy (FTIR). An increased adsorption capacity of Cd was observed for untreated bacteria relative to that for EPS-free bacteria. Surface complexation modeling of titration data showed the similar pK_a values of functional groups (carboxyl, phosphate and hydroxyl) between untreated and EPS-free bacteria. However, site concentrations on the untreated bacteria were found to be higher than those on the EPS-free bacteria. FTIR spectra also showed that no significant difference in peak positions was observed between untreated and EPS-free bacteria and carboxyl and phosphate groups were responsible for Cd adsorption on bacterial cells. The information obtained in this study is of fundamental significance for understanding the interaction mechanisms between heavy metals and biofilms in natural environments.     

城市生活污水中志贺氏菌ipaH毒力基因的定量PCR检测

基于ipaH毒力基因的实时荧光定量PCR检测,建立适合城市污水中志贺氏菌的定量检测方法。使用从临床分离出的志贺氏菌构建重组质粒作为实时荧光定量PCR的标准品。在ipaH基因模板量2.58×100～2.58×106copy范围内具有良好的线性关系,每100 mL水样中含有2.58×101copy以上的ipaH基因即可被检出。从西安市生活污水分离得到2株野生型志贺氏菌,分析ipaH基因数量和菌体数量的关系,从而确定ipaH基因定量检测志贺氏菌的可行性。该方法灵敏、快速、特异性好,适用于城市生活污水中志贺氏菌的检测。     

Mercury biomethylation assessment in the estuary of Bilbao (North of Spain)

The relationship between the microbial methylation of mercury and the microbial activities in sediments and water collected from the estuary of Bilbao (North of Spain) was studied in three different sampling points and in two different seasons. Three different cultures were prepared with a sediment slurry to distinguish between biotic and abiotic methylation pathways and the variations of the methylmercury concentration and the variations of the population of total number of bacteria (TDC), anaerobic heterotrophic bacteria (AHB), sulphate-reducing bacteria (SRB) and Desulfovibrio were measured. From this work, it can be concluded that the variation of MeHg concentrations is a result of the methylation/demethylation processes in the sediments, and that the abiotic processes have a negligible contribution to those processes. According to the statistical analysis of the results (partial least squares analysis) a significant statistical correlation was established between methylmercury and the SRB counts.     

Effects of solids retention time on the performance of bioreactors bioaugmented with a 17β-estradiol-utilizing bacterium, Sphingomonas strain KC8

This study investigated the performance of lab-scale sequencing batch reactors (SBRs) that were inoculated with nitrifying activated sludge and bioaugmented with a Sphingomonas strain KC8 (a 17β-estradiol-degrading bacterium). The bioaugmented SBRs were supplied with synthetic wastewater (average initial total organic carbon (TOC) = 175 mg L⁻¹ and average initial ammonia-N = 25 mg L⁻¹) and daily dose of 17β-estradiol (1 mg L⁻¹) and operated under three solid retention times (SRTs) of 5, 10, and 20 d. After three times periods of the operating SRTs, the overall removal of TOC (>87%) and ammonia (>91%) was similar in all the SBRs. Higher 17β-estradiol removals (>99%) were observed for the SBRs. Neither estrogens nor estrogenic activity was detected in the treated water, except some samples from the SBR operating under 5 d of SRT. The ratios of known estrogen degraders (Sphingomonas strain KC8 and ammonia-oxidizing bacteria) and amoA gene to the total bacterial population decreased as SRT increased, suggesting the presence of unknown estrogen-degraders in SBRs operating at SRT = 10 and 20 d. Real-time-terminal-restriction fragment length polymorphism analysis showed that the evenness of microbial community structures was not affected by the SRT; while, the diversity indices suggest that longer SRTs might lead to more diverse microbial community structure. Overall, the results suggested that bioaugmented bioreactors operating at long SRTs (10 and 20 d) were effective in removing 17β-estradiol to the non-estrogenic treatment endpoint.     

磷酸活化-微波热解改性污泥对镉的吸附性能

以城市污水处理厂剩余污泥为原料,利用磷酸活化-微波热解制取改性污泥。以此污泥作为吸附剂,对含Cd2+废水进行了吸附实验研究。考察了溶液反应时间、Cd2+浓度、pH值和吸附剂用量对镉吸附去除效果的影响;利用等温吸附实验作出吸附等温线,并考察了改性污泥吸附剂吸附Cd2+的动力学方程。实验结果表明,改性污泥对Cd2+有良好的吸附性能,吸附最佳pH值为6.0,吸附较好地符合一级动力学吸附模型和Langmuir-Freundlich等温吸附方程,吸附为物理吸附,吸附反应发生12 h后达到吸附平衡。     

Field and laboratory studies of the fate and enantiomeric enrichment of venlafaxine and O-desmethylvenlafaxine under aerobic and anaerobic conditions

The stereoselectivity of R,S-venlafaxine and its metabolites R,S-O-desmethylvenlafaxine, N-desmethylvenlafaxine, O,N-didesmethylvenlafaxine, N,N-didesmethylvenlafaxine and tridesmethylvenlafaxine was studied in three processes: (i) anaerobic and aerobic laboratory scale tests; (ii) six wastewater treatment plants (WWTPs) operating under different conditions; and (iii) a variety of wastewater treatments including conventional activated sludge, natural attenuation along a receiving river stream and storage in operational and seasonal reservoirs. In the laboratory and field studies, the degradation of the venlafaxine yielded O-desmethylvenalfaxine as the dominant metabolite under aerobic and anaerobic conditions. Venlafaxine was almost exclusively converted to O-desmethylvenlafaxine under anaerobic conditions, but only a fraction of the drug was transformed to O-desmethylvenlafaxine under aerobic conditions. Degradation of venlafaxine involved only small stereoisomeric selectivity. In contrast, the degradation of O-desmethylvenlafaxine yielded remarkable S to R enrichment under aerobic conditions but none under anaerobic conditions. Determination of venlafaxine and its metabolites in the WWTPs agreed well with the stereoselectivity observed in the laboratory studies. Our results suggest that the levels of the drug and its metabolites and the stereoisomeric enrichment of the metabolite and its parent drug can be used for source tracking and for discrimination between domestic and nondomestic wastewater pollution. This was indeed demonstrated in the investigations carried out at the Jerusalem WWTP.     

水质对紫外消毒在两种典型再生水中应用的影响

以大肠杆菌为对象,研究了再生水水质变化对紫外消毒效果和光复活的影响。结果表明,紫外对大肠杆菌有很强的灭活作用,在紫外剂量为4mJ／cm^2时,大肠杆菌的灭活率达到了4．41个对数级。腐殖酸、铁和2种再生水水体中其他溶解性物质会影响光吸收和紫外透射率,但对紫外消毒动力学无影响。在颗粒物浓度为0～200mg／L的范围内,外源高岭土和活性污泥等颗粒物的投加对紫外消毒效果影响较小,而再生水水样W1中原有的2．61／1：g／L的颗粒物则会极大地影响消毒效果,使UV对细菌的灭活出现明显的拖尾现象。腐殖酸会增强紫外损伤大肠杆菌的光复活能力,但2种再生水中细菌的光复活能力相对磷酸盐缓冲溶液（PBS）中减弱,减弱程度在不同水样中有所不同。