Changes of microbial community structures and functional genes during biodegradation of phenolic compounds under high salt condition

The changes of microbial community structures and functional genes during the biodegradation of single phenol and phenol plus p-cresol under high salt condition were explored.It was found that the phenol-fed system (PFS) exhibited stronger degrading abilities and more stable biomass than that of the phenol plus p-cresol-fed system (PCFS).The microbial community structures were revealed by a modern DNA fingerprint technique,ribosomal intergenic spacer analysis (RISA).The results indicated that the microbial community of PFS changed obviously when gradually increased phenol concentration,while PCFS showed a little change.16S rRNA sequence analysis of the major bands showed that Alcanivorax sp.genus was predominant species during phenolic compounds degradation.Furthermore,amplified functional DNA restriction analysis (AFDRA) on phenol hydroxylase genes showed that the fingerprints were substantially different in the two systems,and the fingerprints were not the same during the different operational periods.     

生物滴滤塔处理苯酚气体研究

采用生物滴滤塔处理苯酚气体,考察了苯酚去除性能的影响因素.结果表明,生物滴滤塔能高效处理苯酚气体,苯酚去除效率可达99.5%,长期运行平均去除效率在98%左右.适宜的运行条件为:停留时间20.6 s,循环液pH值7.0,喷淋密度1.67 m3·(m2·h)-1.采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术研究处理苯酚气体的生物滴滤塔填料表面的微生物,结果表明,生物滴滤塔内有5种降解苯酚的优势菌种:Polaromonas sp.、Acinetobacter sp.、Acidovorax sp.、Veillonella parvula和Corynebacterium sp..采用GC-MS分析出口气样,结果表明丙酮酸(CH3COCOOH)为生物降解苯酚的中间产物,并推测了苯酚生物降解的可能途径.     

Kinetics and mechanisms of p-nitrophenol biodegradation by Pseudomonas aeruginosa HS-D38

The kinetics and mecswithhanisms of p-nitrophenol (PNP) biodegradation by Pseudomonas aeruginosa HS-D38 were investigated. PNP could be used by HS-D38 strain as the sole carbon, nitrogen and energy sources, and PNP was mineralized at the maximum concentration of 500 mg/L within 24 h in an mineral salt medium (MSM). The analytical results indicated that the biodegradation of PNP fit the first order kinetics model. The rate constant k_PNP is 2.039 × 10⁻²/h in MSM medium, K_PNP+N is 3.603 × 10⁻²/h with the addition of ammonium chloride and K_PNP+C is 9.74 × 10⁻³/h with additional glucose. The addition of ammonium chloride increased the degradation of PNP. On the contrary, the addition of glucose inhibited and delayed the biodegradation of PNP. Chemical analysis results by thin-layer chromatography (TLC), UV-Vis spectroscopy and gas chromatography (GC) techniques suggested that PNP was converted to hydroquinone (HQ) and further degraded via 1,2,4-benzenetriol (1,2,4-BT) pathway.     

Effective removal of antibiotic resistance genes and potential links with archaeal communities during vacuum-type composting and positive-pressure composting

As a major reservoir of antibiotics, animal manure contributes a lot to the augmented environmental pressure of antibiotic resistance genes (ARGs). This might be the first study to explore the effects of different ventilation types on the control of ARGs and to identify the relationships between archaeal communities and ARGs during the composting of dairy manure. Several ARGs were quantified via Real-time qPCR and microbial communities including bacteria and archaea were analyzed by High-throughput sequencing during vacuum-type composting (VTC) and positive-pressure composting (PPC). The total detected ARGs and class I integrase gene (intI1) under VTC were significantly lower than that under PPC during each stage of the composting (p < 0.001). The relative abundance of potential human pathogenic bacteria (HPB) which were identified based on sequencing information and correlation analysis decreased by 74.6% and 91.4% at the end of PPC and VTC, respectively. The composition of archaeal communities indicated that methane-producing archaea including Methanobrevibacter, Methanocorpusculum and Methanosphaera were dominant throughout the composting. Redundancy analysis suggested that Methanobrevibacter and Methanocorpusculum were positively correlated with all of the detected ARGs. Network analysis determined that the possible hosts of ARGs were different under VTC and PPC, and provided new sights about potential links between archaea and ARGs. Our results showed better performance of VTC in reducing ARGs and potential HPB and demonstrated that some archaea could also be influential hosts of ARGs, and caution the risks of archaea carrying ARGs.     

宏基因组方法分析医药化工废水厂中抗生素耐药菌及耐性基因

废水厂是抗生素耐药菌(ARB)和抗生素耐药基因(ARGs)的巨大储存地.为调查医药化工废水处理厂中的ARB和ARGs,采用了宏基因组技术对医药化工废水中的活性污泥进行取样分析.结果显示,医药化工废水厂微生物组成主要是细菌类,主要细菌门是Proteobacteria,主要属是Hyphomicrobium,主要种是Hyphomicrobium zavarzinii.共检测到74类ARGs,最主要的类型是sav1866、dfr E和mfd.网络分析揭示了ARGs与微分类单元之间的共存模式,即ARGs与废水厂中属级的微生物分类群高度相关.抗生素特异的外排泵是该微生物群落主要的抗生素耐药机制,并且外排泵中耐药结节化细胞分化家族(RND)外排泵占主要部分.该微生物群落最主要的功能通路是代谢相关,并存在许多与人类疾病相关的基因,其中主要是细菌感染性疾病.结果表明,医药化工废水厂蕴藏着丰富的ARB和ARGs,ARGs的累积会增加潜在环境风险,需要加强对医药化工废水厂中ARB和ARGs的监控,并且ARB和ARGs的分析研究对于选择深度处理技术来有效去除ARB和ARGs具有重要的指导意义.     

Characteristics of microbial community functional structure of a biological coking wastewater treatment system

Nitrogenous heterocyclic compounds are key pollutants in coking wastewater; however, the functional potential of microbial communities for biodegradation of such contaminants during biological treatment is still elusive. Herein, a high throughput functional gene array (GeoChip 5.0) in combination with Illumina HiSeq2500 sequencing was used to compare and characterize the microbial community functional structure in a long run (500 days) bench scale bioreactor treating coking wastewater, with a control system treating synthetic wastewater. Despite the inhibitory toxic pollutants, GeoChip 5.0 detected almost all key functional gene (average 61,940 genes) categories in the coking wastewater sludge. With higher abundance, aromatic ring cleavage dioxygenase genes including multi ring1,2diox; one ring2,3diox; catechol represented significant functional potential for degradation of aromatic pollutants which was further confirmed by Illumina HiSeq2500 analysis results. Response ratio analysis revealed that three nitrogenous compound degrading genes- nbzA (nitro-aromatics), tdnB (aniline), and scnABC (thiocyanate) were unique for coking wastewater treatment, which might be strong cause to increase ammonia level during the aerobic process. Additionally, HiSeq2500 elucidated carbozole and isoquinoline degradation genes in the system. These findings expanded our understanding on functional potential of microbial communities to remove organic nitrogenous pollutants; hence it will be useful in optimization strategies for biological treatment of coking wastewater.     

Biodegradation of phenol and /w-cresol by mutated Candida tropicalis

The phenol and m-cresol biodegradations were studied using the mutant strain CTM 2 obtained by the He-Ne laser irradiation on wild-type Candida tropicalis. The results showed that C. tropicalis exhibited the increased capacity of phenolic compounds degradation after laser irradiation. It could degrade 2600 mg/L phenol and 300 mg/L m-cresol by 5% inoculum concentration, respectively. In the dual-substrate biodegradation system, 0-500 mg/L phenol could accelerate m-cresol biodegradation, and 300 mg/L m-cresol could be completely utilized within 46 hr at the presence of 350 mg/L phenol. Besides, the maximum biodegradation of m-cresol could reach 350 mg/L with 80 mg/L phenol within 61 hr. Obviously, phenol, as a growth substrate, could promote CTM 2 to degrade m-cresol, and was always preferentially utilized as carbon source. Comparatively, low-concentration m-cresol could result in a great inhibition on phenol degradation. In addition, the kinetic behaviors of cell growth and substrate biodegradation were described by kinetic model proposed in our laboratory.     

Biodegradation of phenol and m-cresol by mutated Candida tropicalis

The phenol and m-cresol biodegradations were studied using the mutant strain CTM 2 obtained by the He-Ne laser irradiation on wild-type Candida tropicalis. The results showed that C. tropicalis exhibited the increased capacity of phenolic compounds degradation after laser irradiation. It could degrade 2600 mg/L phenol and 300 mg/L m-cresol by 5% inoculum concentration, respectively. In the dual-substrate biodegradation system, 0–500 mg/L phenol could accelerate m-cresol biodegradation, and 300 mg/L m-cresol could be completely utilized within 46 hr at the presence of 350 mg/L phenol. Besides, the maximum biodegradation of m-cresol could reach 350 mg/L with 80 mg/L phenol within 61 hr. Obviously, phenol, as a growth substrate, could promote CTM 2 to degrade m-cresol, and was always preferentially utilized as carbon source. Comparatively, low-concentration m-cresol could result in a great inhibition on phenol degradation. In addition, the kinetic behaviors of cell growth and substrate biodegradation were described by kinetic model proposed in our laboratory.     

Kinetics of phenol and m-cresol biodegradation by an indigenous mixed microbial culture isolated from a sewage treatment plant

An acclimatized mixed microbial culture, predominantly Pseudomonas sp., was enriched from a sewage treatment plant, and its potential to simultaneously degrade mixtures of phenol and m-cresol was investigated during its growth in batch shake flasks. A 2² full factorial design with the two substrates at two different levels and different initial concentration ranges (low and high), was employed to carry out the biodegradation experiments. The substrates phenol and m-cresol were completely utilized within 21 h when present at low concentrations of 100 mg/L for each, and at high concentration of 600 mg/L for each, a maximum time of 187 h was observed for their removal. The biodegradation results also showed that the presence of phenol in low concentration range (100–300 mg/L) did not inhibit m-cresol biodegradation. Whereas the presence of m-cresol inhibited phenol biodegradation by the culture. Moreover, irrespective of the concentrations used, phenol was degraded preferentially and earlier than m-cresol. A sum kinetics model was used to describe the variation in the substrate specific degradation rates, which gave a high coefficient of determination value (R² > 0.98) at the low concentration range of the substrates. From the estimated interaction parameter values obtained from this model, the inhibitory effect of phenol on m-cresol degradation by the culture was found to be more pronounced compared to that of m-cresol on phenol. This study showed a good potential of the indigenous mixed culture in degrading mixed substrate of phenolics.     

Long-term nitrogen and phosphorus removal,shifts of functional bacteria and fate of resistance genes in bioretention systems under sulfamethoxazole stress

To understand the long-term performance of bioretention systems under sulfamethoxazole (SMX) stress, an unplanted bioretention system (BRS) and two modified BRSs with coconut-shell activated carbon (CAC) and CAC/zero-valent-iron (Fe0) granules (CAC-BRS and Fe/CAC-BRS) were established. Both CAC-BRS and Fe/CAC-BRS significantly outperformed BRS in removing total nitrogen (TN)(CAC-BRS:82.48%; Fe/CAC-BRS:78.08%; BRS:47.51%), total phosphorous (TP)(CAC-BRS:79.36%; Fe/CAC-BRS:98.26%; BRS:41.99%),and...     

渔业复垦塌陷地抗生素抗性基因与微生物群落

水产养殖行业抗生素的广泛使用引起了抗生素抗性基因（antibiotic resistance genes,ARGs）污染问题.为探究渔业复垦塌陷地ARGs污染特征,利用宏基因组学技术检测分析了渔业复垦塌陷地ARGs的相对丰度和微生物群落结构.研究区域共检测出29种ARGs,bacA在所有样品中相对丰度最高,达1.96×10^-5~1.19×10^-4.沉积物中磺胺类和四环素类ARGs相对丰度较高,井水中多药类ARGs相对丰度较高.微生物群落结构表明变形菌门（Proteobacteria）在所有样品中为最优势细菌门,沉积物中绿弯菌门（Chloroflexi）和广古菌门（Euryarchaeota）较为优势.属水平上,硫杆菌属（Thiobacillus）为沉积物中最优势细菌属,不动杆菌属（Acinetobacter）和假单胞菌属（Pseudomonas）为井水中优势细菌属.ARGs与微生物相关性分析表明,菌属与ARGs间主要呈中度相关,多种菌属与ARGs显著正相关,ARGs的分布受微生物群落结构的重要影响.渔业复垦塌陷地沉积物与井水均受到ARGs污染,应加强相应控制措施保护区域环境.     

恶臭假单胞菌（Pseudomonas putida LY1）共代谢降解苯酚和4-氯苯酚系统的降解动力学研究

对恶臭假单胞菌（Pseudomonas putida LY1）共代谢降解苯酚和4-氯苯酚（4-CP）系统进行了降解实验和动力学研究.结果表明,恶臭假单胞菌可以有效地降解苯酚,苯酚浓度为50mg·L-1时细菌生长速度最快,4-氯苯酚浓度的增加会对细菌产生一定的抑制作用.同时,用改进的Haldane方程模拟恶臭假单胞菌LY...     

磺胺抗性消长与堆肥进程的交互特征

基于添加和不添加磺胺药[m(磺胺二甲嘧啶SM2)∶m(磺胺-6-甲氧嘧啶SMM)=1∶1]的好氧堆肥试验,分析堆肥过程中鸡粪理化性质、微生物群落代谢特征、磺胺抗生素以及5种抗性基因变化,探明磺胺抗性消长与堆肥过程的交互特征.结果表明,磺胺药添加抑制了堆肥基础呼吸,延长了堆体达到高温的时间,减缓了养分转化速度,显著影响堆肥中期微生物群落结构特征.鸡粪中SMM和SM2在堆肥14 d内即可完全降解,且SMM降解速率高于SM2.随堆肥进行,sul1和sul2呈先下降后轻微回升的趋势,磺胺药添加对sul1和Int I1丰度无明显提高作用,但可促进sul2扩散.堆肥过程中,tet Q和tet W变化与磺胺抗性基因不同,但磺胺药添加亦增加tet Q和tet W相对丰度.冗余分析结果显示,温度与sul1、sul2和Int I1明显负相关,与tet Q和tet W无明显相关性;5种抗性基因相对丰度均与碳氮比和硝态氮含量负相关,与p H、含水率和铵态氮含量正相关.     

Photoelectrochemical performance of birnessite films and photoelectrocatalytic activity toward oxidation of phenol

Birnessite films on fluorine-doped tin oxide(FTO) coated glass were prepared by cathodic reduction of aqueous KMnO_4. The deposited birnessite films were characterized with X-ray diffraction, Raman spectroscopy, scanning electron microscopy and atomic force microscopy.The photoelectrochemical activity of birnessite films was investigated and a remarkable photocurrent in response to visible light was observed in the presence of phenol, resulting from localized manganese d–d transitions. Based on this result, the photoelectrocatalytic oxidation of phenol was investigated. Compared with phenol degradation by the electrochemical oxidation process or photocatalysis separately, a synergetic photoelectrocatalytic degradation effect was observed in the presence of the birnessite film coated FTO electrode.Photoelectrocatalytic degradation ratios were influenced by film thickness and initial phenol concentrations. Phenol degradation with the thinnest birnessite film and initial phenol concentration of 10 mg/L showed the highest efficiency of 91.4% after 8 hr. Meanwhile, the kinetics of phenol removal was fit well by the pseudofirst-order kinetic model.     

固定化微生物技术在难降解有机污染物治理中的研究进展

综述了90年代以来固定化微生物技术用于难降解有机污染物治理中的最新研究进展,这些难降解有机污染物包括:酚类(苯酚、氯代酚、甲酚、硝基酚等),芳香烃类(单环芳烃、多环芳烃、杂环芳烃)及其他化合物(酰胺类、氰、农药、杀虫剂、合成洗涤剂等)。并对固定化技术的应用前景及存在的问题进行了评述。      

好氧-厌氧两相堆肥过程中抗生素耐药基因的变化特征及影响因素研究

针对堆肥处理过程中抗生素耐药基因（antibiotic resistance genes,ARGs）去除效率不高、诱导潜在致病菌传播扩散等环境安全问题,采用好氧-厌氧两相堆肥处理猪粪、秸秆,探讨ARGs丰度及微生物群落的变化特征,并明确影响ARGs相对丰度变化的关键因子.结果表明:与单一好氧相比,好氧-厌氧两相堆肥能更有效地降低部分ARGs及潜在致病菌的相对丰度,其中ermB、tetK的相对丰度分别降低了39.68%、72.19%,假单胞菌属（Pseudomonas）、梭菌属（Clostridium_sensu_stricto_1）的相对丰度分别降低了96.09%、90.07%.主成分分析（Principal Component Analysis,PCA）及冗余分析（Redundancy analysis,RDA）得出,微生物群落和环境因子均显著影响ARGs相对丰度的变化（P < 0.05）,贡献率分别为82.0%、71.2%.其中,梭菌属（Clostridium_sense_stricto_1）、黄杆菌属（Flavobacterium）、尿素芽孢杆菌属（Ureibacillus）和海洋杆菌属（Oceanobacillus）是影响ARGs相对丰度变化的主要微生物菌属;温度、电导率、含水率和pH是影响ARGs相对丰度变化的关键环境因子,贡献率分别为22.0%、16.7%、16.2%和15.0%.研究显示:与好氧堆肥相比,好氧-厌氧两相堆肥能更有效地去除ARGs,并抑制潜在致病菌的繁殖扩散,从而减少环境风险;然而部分ARGs和潜在致病菌仍存在增殖现象,因此亟需进一步优化好氧-厌氧两相堆肥工艺.      

长期施用猪粪水稻土抗生素抗性基因污染研究

为了全面研究猪粪有机肥对水稻土抗生素抗性及分布格局的影响,选择295对抗生素抗性基因引物,采用高通量荧光定量PCR对水稻土中的抗生素抗性基因污染情况进行了研究.结果表明,未施用猪粪水稻土检测出66种抗性基因,而施用猪粪水稻土则检测出107种抗生素抗性基因,施用猪粪后水稻土中抗生素抗性基因种类显著增加(P<0.05).相对于未施用猪粪水稻土,施用猪粪水稻土有49种抗生素抗性基因丰度显著增加(P<0.05),其中施用猪粪水稻土中喹诺酮类/氯霉素类抗性基因的mexF的丰度相对于未施用猪粪水稻土增加1791倍.高通量定量结果表明,施用猪粪的水稻土抗生素抗性基因分布格局发生显著变化,高通量定量技术是研究环境抗生素抗性基因的一个非常实用的工具.     

基于宏基因组技术分析MBR膜清洗后污泥中抗性基因

污水处理厂作为抗生素抗性基因（antibiotic resistance genes,ARGs）的重要储存库,是自然界ARGs的主要来源之一.膜生物反应器（membrane bioreactor,MBR）被认为是一种能够有效去除污水处理厂中ARGs的技术工艺.MBR膜截留的废水中胶体、颗粒物、悬浮物及微生物代谢物中存在着大量的病原菌与抗性基因,而目前关于膜清洗后污泥中抗性基因的分布特征和规律尚不明确.本文采用宏基因组技术对MBR膜清洗后污泥中抗性基因进行了分析.结果显示,膜清洗后污泥中共检测出39门,其中优势菌门为Proteobacteria、Nitrospirae和Actinobacteria,优势菌属为Nitrospira、Pseudomonas和Bradyrhizobium.污泥样品含有的病原菌属占所有菌属的10.54%,其中Pseudomonas属相对丰度最高,占到所有菌属的3.94%.样品中共注释出17类ARGs和16类金属抗性基因（metal resistance genes,MRGs,15类单金属抗性基因和1类多重金属抗性基因）.其中,多药类抗生素抗性基因相对丰度最高,占49.08%.金属抗性基因中多重金属类抗性基因相对丰度最高,占该污泥样品的34.58%,单金属抗性基因中对铜的抗性基因数量最多,占19.99%.该膜清洗后污泥中微生物群落最主要的功能通路为代谢相关,并存在大量与人类疾病相关的代谢通路相关基因,其中涉及细菌耐药和细菌传染疾病的基因数量最多,分别为占人类疾病相关的代谢通路已注释序列的34.50%和16.62%.由此可见,膜清洗后污泥中蕴藏着丰富的ARGs、MRGs以及病原菌属,具有潜在的环境健康风险,需要加强对膜清洗后污泥中ARGs、MRGs以及病原菌的管控.本文为选择合适的技术工艺有效去除膜清洗后污泥中ARGs、MRGs以及病原菌提供指导.     

Effect of temperature switchover on the degradation of antibiotic chloramphenicol by biocathode bioelectrochemical system

Exposure to chloramphenicol(CAP),a chlorinated nitroaromatic antibiotic,can induce CAP-resistant bacteria/genes in diverse environments. A biocathode bioelectrochemical system(BES) was applied to reduce CAP under switched operational temperatures.When switching from 25 to 10°C,the CAP reduction rate(kCAP) and the maximum amount of the dechlorinated reduced amine product(AMCl,with no antibacterial activity) by the biocathode communities were both markedly decreased. The acetate and ethanol yield from cathodophilic microbial glucose fermentation(with release of electrons) was also reduced. Formation of the product AMCl was enhanced by the biocathode dechloridation reaction compared with that produced from pure electrochemical or microbial dechloridation processes. The electrochemical and morphological analyses of cathode biofilms demonstrated that some cathodophilic microbes could adapt to low temperature and play a key role in CAP degradation. The resilient biocathode BES has a potential for the treatment of CAP-containing wastewater in temperature fluctuating environments.     

活性炭对城市有机固废厌氧消化过程抗生素抗性基因行为特征的影响

城市有机固废是抗生素抗性基因（antibiotic resistance genes,ARGs）的来源和储存库之一,其生物处理过程中ARGs的赋存变化规律需深入研究.采用定量PCR方法分析城市有机固废厌氧消化过程中多种类型ARGs和整合子基因的变化特征,探究了不同粒径的活性炭对目标基因行为特征的影响以及ARGs变化的潜在微生物机制.结果表明,无论是否添加活性炭,厌氧消化过程对初始体系中的总ARGs均具有削减作用,总ARGs绝对丰度的削减率为29.95%~63.40%.城市有机固废厌氧消化最终体系中,粉末活性炭（powered activated carbon,PAC）添加组中总ARGs丰度显著高于对照组（P<0.05）,即PAC削弱了ARGs的削减效果,颗粒活性炭对ARGs变化无显著影响.厌氧消化过程中,ARGs的潜在宿主细菌主要属于梭菌纲（Clostridia）、拟杆菌纲（Bacteroidia）和互营养菌纲（Synergistia）.PAC添加时,潜在宿主细菌的富集是目标基因增殖的重要原因,且Clostridia可能是厌氧消化过程中ARGs消长的主要驱动因子.本研究结果将为了解城市有机固废厌氧消化过程中ARGs的转归特征以及外源添加活性炭对ARGs的影响机制提供参考.