抗生素抗性异养细菌在不同水体中的生态分布研究

比较了城市污水处理厂曝气池混合液、鱼塘养殖水和黄河引灌水3种水体中可培养异养细菌总数、不同抗生素抗性菌群比例以及抗性有色异养细菌的比例.3种水体中可培养异养细菌总数分别达到2.8×107、1.2×107和8.9×105CFU/mL,其中,污水处理厂曝气池混合液中青霉素、红霉素和链霉素抗性比例最高,分别达到84.5%、66.1%和62.5%,明显高于另外2种水体中抗性比例,而且在该水体中检测到了102～104CFU/mL对青霉素、链霉素和庆大霉素不同组合的交叉抗性菌群.另外,在所有含抗生素的培养基平板上,有色菌群(黄色和红色)的比例都明显高于相应的未加抗生素的培养基平板,说明抗生素抗性与细菌的色素积累呈正相关,其中,黄河引灌水中的抗性有色异养菌群的比例高于其他2种水体.     

城市污水处理厂污水处理系统中四环素类药物耐药菌的分布特性研究

采用微生物平板涂布法对2家城市污水处理厂污水处理各阶段的四环素类药物(四环素、金霉素、土霉素)耐药菌分布进行了测定,分析了污水处理的各阶段对四环素类药物耐药菌的去除效果。结果显示,2家污水处理厂污水处理系统中均能检出土霉素、四环素、金霉素耐药菌,数量分布均为土霉素耐药菌>四环素耐药菌>金霉素耐药菌。3种耐药菌的浓度均在加氯消毒出水中为最小(在0～103 cfu/mL),在外排污泥中最大(高于进水浓度,在105～107 cfu/mL)。四环素耐药菌数从进水的1.1×105～1.6×105 cfu/mL降为出水的0～5.5×102 cfu/mL,说明污水处理过程对四环素耐药菌有一定的去除效果,但四环素耐药菌占总菌数的百分比没有明显下降或上升,相比于进水的1.1%～3.4%,出水仍然保持在3.1%～4.3%左右,表明污水处理过程中没有选择性的富集或去除四环素耐药菌。     

北京市某城市污水处理厂微生物气溶胶污染特性研究

以北京市某城市污水处理厂的格栅间、曝气池、污泥浓缩池和污泥脱水间为对象,研究不同功能区的空气微生物浓度、粒径分布及微生物组成.结果表明:(1)该污水处理厂4个功能区空气异养细菌浓度平均值为3.3×104cfu/m3,范围为1.0×102～4.3×105cfu/m3;真菌浓度平均值为6.1×103cfu/m3,范围为7.5×10～7.6×104cfu/m3.异养细菌和真菌浓度存在显著性差异,尤以曝气池上空最高,其次是污泥脱水间,浓度最低的区域为格栅间和污泥浓缩池.各功能区均存在不同程度的异养细菌和真菌污染.(2)该污水处理厂不同功能区逸散出的异养细菌分布比例最高的为第2～5级,真菌主要分布在第3～5级.4个功能区空气中粒径为2.10～4.70μm(第3、4级)的异养细菌和真菌粒子分别占到了总数的37％～40％和46％～56％,而粒径为0.65～2.10μm(第5、6级)的异养细菌和真菌粒子则分别占到了总数的30％～33％和31％～37％,说明该污水处理厂不同功能区内的空气微生物存在一定的健康风险,可能对人呼吸道构成感染威胁.(3)假单孢菌(Pseudomonas)为该污水处理厂不同功能区均检出的优势异养细菌,而优势真菌种属为毛霉(Mucor)和曲霉(Aspergillus).     

生物强化处理石油污染土壤理化性质和微生物学特性的纵向分布特征

采用原位强化生物修复技术对某区块石油污染土壤进行为期16个月的生物修复,考察了处置后污染土壤理化性质、微生物学特性以及石油烃组成的纵向分布特征。实验结果表明,经过修复后各土层的石油烃去除率是表层土IN-3(50.42%)中层土IN-2(23.54%)底层土IN-1(10.51%);IN-1处于缺氧环境,存在硫酸盐还原和反硝化作用,使得土壤pH值从7.86±0.03降低至7.27±0.03,土壤总氮从2.53±0.13 g/kg降低至0.77±0.04 g/kg;厌氧菌的种群数量是IN-1(10.43±0.71×104CFU/g)IN-3(6.74±0.39×104CFU/g)IN-2(5.15±0.42×104CFU/g),放线菌数量与石油烃含量显著负相关(r=-0.989,p=0.0110.05);IN-3对饱和份和芳香份的降解率最高,分别达到了70.27%和54.52%,远高于IN-2和IN-1;模拟蒸馏结果表明,IN-3正构烷烃得到了很大程度的去除,缺氧的IN-1对正构烷烃去除得较少;厌氧菌数量与胶质和沥青质去除率之间成正相关关系,对于污染源较为分散的污染区域,采用原位生物强化修复时可以考虑引入厌氧修复。     

不同畜禽养殖场中气载抗生素抗性基因的污染特征与季节性变化

由于大量的抗生素被用于畜禽养殖行业,畜禽养殖场中的抗生素抗性基因(ARGs,Antibiotic resistance genes)问题正逐渐变得严重。为调查畜禽养殖场中气载ARGs的污染特征及季节性变化,分别在冬季和夏季采集陕西省渭南市养猪场和养鸡场的空气和粪便样品,分析不同种类气载ARGs的污染水平及ARGs与环境因子的相关性。结果表明：畜禽养殖场空气中主要检出的ARGs类型为万古霉素类抗性基因、MLSB类抗性基因、四环素类抗性基因、FCA类抗性基因;在不同类型养殖场中气载ARGs总浓度趋势为养鸡场>养猪场,且养鸡场中每种ARGs浓度比养猪场中高约2个数量级;在不同季节养殖场中,气载ARGs总浓度趋势为冬季>夏季,且每种ARGs在冬季均高于夏季2～3个数量级。Pearson相关性分析结果表明,养殖场中气载ARGs总浓度与PM2.5呈显著相关(P<0.05),与其他环境因子无显著相关性。本研究成果可为控制畜禽养殖场中气载ARGs污染的传播及防控提供参考。     

水中抑制物对定量PCR与膜过滤法检测E.coli的影响

针对水中病原微生物的污染,选择大肠埃希氏菌E.coli作为病原示踪剂,以E.coli染色体上β-葡萄糖醛酸酶uid A目的基因建立了SYBR Green实时荧光定量PCR的检测方法。该定量PCR方法灵敏度高,检测限可达104CFU/L,线性关系良好,相关系数为R2=0.999。研究表明,定量PCR与菌液浓度呈显著正相关R2=0.935。通过人工投加腐殖酸、COD,研究了水中抑制物对定量PCR和膜过滤MF培养法的影响。结果显示,腐殖酸可使E.coli在培养皿上的菌落变小聚集,且显色不明显,当腐殖酸量为20 mg/L时,浓度为50 CFU/100 m L的E.coli完全受到抑制,没有菌落出现。腐殖酸对PCR扩增的抑制作用明显,当腐殖酸浓度增加到10 mg/L,定量PCR检测结果基因拷贝数减少约1 log,增加到20 mg/L定性PCR检测结果为阴性。     

生活污水中肠道病毒分布特性的定量PCR研究

为了明确污水中肠道病毒的分布及去除规律,采用定量PCR方法对西安市3个污水厂进行了为期一年的监测。结果表明,肠道病毒和人类肠道病毒在进水和二级出水中呈较高浓度,约分布在102~105copies/m L和102~104copies/m L范围内。肠道病毒和人类肠道病毒浓度呈现季节性变化,秋季浓度最高,冬季次之,春夏最低。经统计学分析发现,肠道病毒和人类肠道病毒浓度在3个水厂间无显著差异性(p0.05)且肠道病毒和人类肠道病毒很好地服从对数正态分布。通过病毒的检测数据及分布特性,明确了函数参数并确定人类肠道病毒占肠道病毒的39%。经过常规二级处理,肠道病毒可去除1.12 log左右,与人类肠道病毒去除率相近,且不同水厂对病毒的去除率也相近。     

堆肥对污泥中四环素类抗生素及抗性基因的影响

近年来,抗生素及抗性基因的污染与扩散成为全球关注的热点。污泥,作为污水处理系统的副产物,是环境中抗生素抗性基因的重要存储库。污泥经堆肥后可作为肥料施用于农田,然而关于污泥堆肥过程中抗性基因丰度变化的研究却很少。对污泥堆肥过程中四环素类抗生素(四环素、土霉素和金霉素)和四环素类抗性基因(tet(A),tet(C),tet(M),tet(O)和tet(X))的动态变化进行了定量研究。结果表明,四环素、土霉素和金霉素的浓度经堆肥处理后分别减少85.6%,91.4%和85.3%。高温是导致四环素类抗生素降解的主要因素。经堆肥处理后,tet(A)和tet(X)的相对丰度增加,tet(C)、tet(M)和tet(O)的相对丰度略微下降,说明堆肥处理对污泥中四环素类抗性基因的削减效果有限。     

粉末活性炭-超滤组合工艺对二级出水中污染物的去除及膜污染机制研究

采用粉末活性炭(PAC)-超滤(UF)组合工艺处理污水处理厂二级出水,探讨不同PAC投加量下PAC-UF组合工艺对溶解性有机物(DOC)及抗生素抗性基因(ARGs)的去除效果,分析PAC对UF膜污染的缓解机制。结果表明,与直接UF相比,PAC-UF组合工艺可有效降低出水DOC和ARGs含量;水中4种ARGs与微生物含量、整合子intI1、DOC浓度间呈显著相关关系,说明去除上述指标有助于削减不同类型ARGs;PAC可吸附水中小分子量有机物,提高膜比通量,改善UF膜的反冲洗效果,PAC投加量为20mg/L时效果最好;PAC投加量增加可使滤饼层变得致密,使UF膜的不可逆污染阻力下降,但总污染阻力增加;直接UF与PAC-UF组合工艺的膜污染主导机制均为滤饼层污染,其中PAC-UF组合工艺受滤饼层污染机制影响更大。综合考虑污染物去除及膜污染缓解效果,采用低投加量(20mg/L)的PAC-UF组合工艺处理二级出水最为适宜。     

嗜热栖热菌降解氟喹诺酮类抗生素

氟喹诺酮类抗生素在各种环境基质中积累造成的生态和耐药基因污染等问题已引起广泛的关注。为了能够有效去除环境中氟喹诺酮类抗生素污染并且探究其生物代谢途径,利用嗜热菌Thermus sp. C419在高温(70℃)条件下降解2种典型的氟喹诺酮类抗生素(诺氟沙星和恩诺沙星),分析了菌株C419对这2种药物在单一和混合添加时的降解特性;通过UPLC-MS/MS检测了其相关的降解产物,并推测了可能的代谢途径;利用平板扩散法对生物降解后的氟喹诺酮类药物进行抑菌活性测定。结果表明:氟喹诺酮类化合物可被菌株C419有效降解,降解率为60%～80%;该生物降解过程符合一级动力学模型,培养基中氟喹诺酮类化合物浓度越高,降解率越高,降解半衰期越短;菌株C419对诺氟沙星的生物降解有3条可能的降解途径和7种降解产物,对恩诺沙星的生物降解有4条可能的降解途径和6种降解产物。此外,与2种药物的母体化合物相比,生物降解后药物对不同细菌的抗菌活性均有一定程度的降低,这说明嗜热菌株C419在热环境中去除氟喹诺酮类污染物方面可能会具有良好的实用性和应用前景。     

MSMA resistance studies

Monosodium methanearsonate (MSMA)-resistant and -susceptible common cocklebur (Xanthium strumarium L.) and cotton (Gossypium hirsutum L.) were treated with MSMA. Plant parameters analyzed were: glutathione synthetase activity, selected amino acid (arginine, glutamic acid, alanine, citrulline, glutamine, and glutathione) content and arsenic content (MSMA, total arsenic, and arsonate). No reduction of arsenic from the parent pentavalent form present in MSMA to the trivalent form was detected. Arginine, glutamic acid, and glutamine content increased in tissue three days after MSMA treatment. Glutathione content decreased during the first three days after treatment; however, five days after treatment the resistant biotype of cocklebur and cotton had elevated glutathione levels (8-20 times greater, respectively). Glutathione Synthetase activity was higher in cotton than in either of the cocklebur biotypes; MSMA did not affect its activity in cotton or either cocklebur biotype. Resistant biotypes have a slightly higher activity than the susceptible biotype. Tolerance of cotton to MSMA may be related to glutathione synthetase activity and possibly to the presence of phytochelatins. Also, increased glutathione levels in the resistant biotype may implicate phytochelatin involvement in the resistance mechanism.     

人工湿地中抗生素抗性大肠杆菌和抗性基因的去除与分布

抗生素的滥用导致抗生素抗性菌和抗性基因随生活污水和养殖废水的排放在环境中肆意散播,其去除及环境行为越来越受到关注。采用K-B纸片法测定了9套不同工艺构型模拟人工湿地中大肠杆菌对7种抗生素的抗性率,并应用多重PCR检测磺胺类sul1、2、3与四环素tetA、B、C、D抗性基因,探究人工湿地对抗性菌的去除效率及抗性菌、抗性基因的分布规律。结果显示,人工湿地能有效去除污水中70%左右的抗性大肠杆菌,有效降低了细菌抗性的传播风险;共计分离出535株大肠肝菌中有378株对一种以上抗生素有抗性性,以四环素、磺胺类和氨苄西林抗性率最高,达到25%以上,其他4种抗性率较低,不足20%;2种抗性基因的检出率都在70%以上;对不同采样点大肠杆菌的抗性性及抗性基因的比较发现,各部分大肠杆菌的抗性水平、多重抗性指数(MRI)以及抗性基因(sul、tet)检出率和组合数表现出:基质≥出水>进水,推测抗性菌被湿地基质截留,在基质生物膜上发生抗性基因的重组,并释放抗性菌,提高了出水中抗性水平和抗性基因检出率。     

植物抗逆性研究进展

本文论述了逆境对植物各方面造成的伤害及植物产生的相应反应 ,重点对植物在生理生化上的变化、对抗性生理领域中的一些学说和最新研究结果作了介绍 ,并指出相生相克现象也可作为抗逆生理的一个方面加以研究 ,概念扩展后的抗逆生理学必将在农业、环保等方面得到更广泛的应用     

Detection of antibiotic resistance and tetracycline resistance genes in Enterobacteriaceae isolated from the Pearl rivers in South China

This study investigated antibiotic resistance profiles and tetracycline resistance genes in Enterobacteriaceae family isolates from the Pearl rivers. The Enterobacteriaceae isolates were tested for susceptibility to seven antibiotics ampicillin, chloramphenicol, ciprofloxacin, levofloxacin, sulphamethoxazole/trimethoprim, tetracycline and trimethoprim. In Liuxi reservoir, with an exception to ampicillin resistant strains (11%) no other antibiotic resistance bacterial strains were detected. However, multiple drug resistance in bacterial isolates from the other sites of Pearl rivers was observed which is possibly due to sewage discharge and input from other anthropogenic sources along the rivers. Four tetracycline resistance genes tet A, tet B, tet C and tet D were detected in the isolates from the rivers. The genes tet A and tet B were widely detected with the detection frequencies of 43% and 40% respectively. Ciprofloxacin and levofloxacin resistant enteric bacteria were also isolated from the pig and duck manures which suggest a wider distribution of human specific drugs in the environment. This investigation provided a baseline data on antibiotic resistance profiles and tetracycline resistance genes in the Pearl rivers delta.     

Natural resistance of rose petals to microbial attack

Petals of red, yellow and white roses (Rosa damascene Mill.) of the family Rosaceae were extracted with (1:1) methylene chloride/methanol and tested for their antimicrobial activities against four species of Gram-positive bacteria (Bacillus cereus, Bacillus subtilis, Micrococcus luteus and Staphylococcus aureus), five species of Gram-negative bacteria (Enterobacter aerogenes, Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa and Serratia marcescens) and five species of fungi (Penicillium notatum, Aspergillus niger, Rhizopus stolonifer, Saccharomyces cerevisiae and Fusarium oxysporum). All of the crude extracts showed a wide range of antimicrobial activities according to the tested organism and rose's type. Micrococcus luteus was found to be the most susceptible bacteria to all crude extracts. Red and yellow petal extracts showed much higher antibacterial activity than the white petals extract. Bacillus subtilis was found to be the least susceptible to all extracts. The fungus, Penicillium notatum was found to be the most susceptible with white petal extract being the most effective. Saccharomyces cerevisiae and Fusarium oxysporum were the least susceptible to all extracts. White roses extract showed much higher antifungal activities against Penicillium notatum than red or yellow roses, therefore, it was subjected to several bioassay guided chromatographic fractionations and purification to isolate the active chemical(s) responsible for the antifungal activity. Chemical structure of the isolated antifungal compounds were identified by spectroscopy techniques and found to be a γ-sitosterol and (Z,Z)-9,12-octadecadienoic acid. Antibacterial activity of the various types of rose extracts were due to complex mixtures of organic compounds which are still under chemical investigation and will be published later.     

Cornmeal-induced resistance to ciprofloxacin and erythromycin in enterococci

Triclosan is used as an antibacterial agent in household items and personal care products. Since this compound is found in maternal milk of humans and bodies of wild animals, there is growing concern among some consumer groups and scientific community that triclosan is adverse for humans and wild animals. In order to estimate adverse actions of triclosan, the effects of triclosan on intracellular Zn²⁺ concentration and cellular thiol content were studied in rat thymocytes by the use of flow cytometer with appropriate fluorescent probes. Triclosan at 1-3 μM (sublethal concentrations) increased the intensity of FluoZin-3 fluorescence (intracellular Zn²⁺ concentration) and decreased the intensity of 5-chloromethylfluorescein (5-CMF) fluorescence (cellular thiol content). Negative correlation (r = −0.985) between triclosan-induced changes in FluoZin-3 and 5-CMF fluorescences was found. Removal of external Zn²⁺ did not significantly affect the triclosan-induced augmentation of FluoZin-3 fluorescence, suggesting an intracellular Zn²⁺ release by triclosan. These actions of triclosan were similar to those of H₂O₂ and triclosan significantly potentiated the cytotoxicity of H₂O₂. Therefore, the results may suggest that triclosan at sublethal concentrations induces oxidative stress that decreases cellular thiol content, resulting in an increase in intracellular Zn²⁺ concentration by Zn²⁺ release from intracellular store(s). Since recent studies show many physiological roles of intracellular Zn²⁺ in cellular functions, the triclosan-induced disturbance of cellular Zn²⁺ homeostasis may induce adverse actions on the cells.     

Natural resistance of rose petals to microbial attack

Petals of red, yellow and white roses (Rosa damascene Mill.) of the family Rosaceae were extracted with (1:1) methylene chloride/methanol and tested for their antimicrobial activities against four species of Gram-positive bacteria (Bacillus cereus, Bacillus subtilis, Micrococcus luteus and Staphylococcus aureus), five species of Gram-negative bacteria (Enterobacter aerogenes, Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa and Serratia marcescens) and five species of fungi (Penicillium notatum, Aspergillus niger, Rhizopus stolonifer, Saccharomyces cerevisiae and Fusarium oxysporum). All of the crude extracts showed a wide range of antimicrobial activities according to the tested organism and rose's type. Micrococcus luteus was found to be the most susceptible bacteria to all crude extracts. Red and yellow petal extracts showed much higher antibacterial activity than the white petals extract. Bacillus subtilis was found to be the least susceptible to all extracts. The fungus, Penicillium notatum was found to be the most susceptible with white petal extract being the most effective. Saccharomyces cerevisiae and Fusarium oxysporum were the least susceptible to all extracts. White roses extract showed much higher antifungal activities against Penicillium notatum than red or yellow roses, therefore, it was subjected to several bioassay guided chromatographic fractionations and purification to isolate the active chemical(s) responsible for the antifungal activity. Chemical structure of the isolated antifungal compounds were identified by spectroscopy techniques and found to be a γ-sitosterol and (Z,Z)-9,12-octadecadienoic acid. Antibacterial activity of the various types of rose extracts were due to complex mixtures of organic compounds which are still under chemical investigation and will be published later.     

无机微滤膜过滤阻力的研究

本文采用孔径为 0 .8μm、0 .2 μm和 5 0nm的无机微滤膜 ,考察了不同情况下膜的过滤阻力分布情况。对于造纸黑液 ,当浓度较低时 ,过滤阻力由膜阻力、吸附阻力、堵孔阻力和浓差极化阻力共同控制 ;当浓度较高时 ,吸附阻力起主要作用。过滤 1g/L聚乙二醇溶液的试验结果表明 ,不同孔径膜的过滤阻力构成是不同的 ,孔径越小则吸附阻力越小 ,但浓差极化阻力会显著增加     

可张孔曝气器曝气能耗试验研究

通过试验研究，总结出可张孔曝气阻力损失的影响因素，以优化产品的结构设计参数，降低产品在工程应用中的能耗，获取更好的经济效益。