环境抗生素污染的微生物修复进展

近年来随着抗生素在畜牧业、水产养殖业以及医疗行业的广泛应用,大量抗生素通过排泄物进入环境,导致我国大面积水体及土壤环境中抗生素残留量急剧增高。环境中不同种类的抗生素的残留导致微生物种群结构失衡,对生态环境及人类造成极大危害。因此,解决抗生素残留问题是21世纪新型环境污染物领域的一个重要课题。已有研究显示,一些微生物能够以抗生素为碳源生存,可用于降解环境中残留抗生素,但人们对微生物降解抗生素的降解机制了解较少。文中概括了近十年来抗生素降解菌株和菌群对抗生素的去除情况,以及应用微生物菌群处理抗生素残留的技术方法,同时对未来利用微生物修复法减少环境中抗生素残留进行了展望。     

土壤环境中典型抗生素残留及其与微生物互作效应研究进展

抗生素大量不规范使用,其残留对环境和人类健康造成严重威胁。污水灌溉和生物有机肥的施用使土壤成为抗生素的主要归趋地之一,抗生素残留对土壤微生物产生不同程度的影响。首先综述了土壤抗生素污染现状,进一步论述了土壤抗生素残留对抗性基因诱导和微生物群落结构的影响,重点介绍了微生物方法在去除抗生素污染方面的重要作用,并指出了土壤抗生素污染研究中存在的问题,对今后主要研究方向进行展望。     

4种市售抗生素对发酵床垫料中益生微生物生长的影响

目的研究不同剂量土霉素、盐酸环丙沙星、克拉霉素、乙酰甲喹对垫料中益生微生物生长的影响。方法以饲料中抗生素添加量、抗生素在动物体内转化率为依据,观察0.1×国标、1×国标和10×国标对应的抗生素残留对垫料中主要益生微生物生长的影响。结果与空白组相比,0.1×国标的土霉素和克拉霉素残留对3种益生微生物影响小,盐酸环丙沙星和乙酰甲喹残留使枯草芽胞杆菌和酵母菌数量不可检测;1×国标量的土霉素残留对3种益生微生物影响较小,而盐酸环丙沙星、克拉霉素、乙酰甲喹残留造成了益生微生物的不可检测;除植物乳杆菌在10×国标的土霉素残留中生长变化小外,所有检测菌在该浓度其他抗生素残留中均不可检测。结论土霉素、盐酸环丙沙星、克拉霉素、乙酰甲喹残留对垫料中主要益生微生物生长均有不同程度的影响。     

土壤中抗性基因的产生,扩散传播以及消减的研究进展

近年来,土壤中残留的大量抗生素不可避免的导致耐药微生物和抗性基因的增加和扩散,引起一系列土壤污染和生态风险。作为一类新兴污染物,抗性基因的污染水平已经远远超出我们的预想,因此对土壤中抗性基因的分布水平、扩散传播及消减技术的研究刻不容缓。本文对国内外土壤中抗生素和抗性基因残留水平进行了总结分析,探讨了土壤中抗性基因的产生、扩散的内在动力和机制。同时,分析了土壤中抗性基因分布和扩散的影响因素,如:抗生素残留水平,土壤理化性质和环境条件等。在此基础上,探讨了土壤抗性基因阻隔和消减技术,包括传统降解方法:高温,光照催化、微波-H2O2-微生物联合处理技术等,并提出新型消解技术:取代活性基团、靶位修饰以及改变外排泵的通透性等。讨论未来在控制抗性基因生态风险,降低其在土壤中的丰度,有效阻截技术的发展趋势。     

微生物学检测技术在食品抗生素残留检测的研究应用

抗生素的普遍应用使得其在食品中广泛残留,对人体健康造成严重危害。目前,用于食品抗生素残留检测方法较多,微生物法是常用的筛选方法之一,具有简便、经济、高通量、特异性及灵敏度好等优点。本研究综述了微生物法在检测食品中抗生素残留的优缺点和影响因素等应用情况。     

环境介质中的抗生素及其微生物生态效应

环境介质中的抗生素因存在浓度较低被称为微量污染物,其对生态系统和人类健康的影响已逐步得到认知。长期以来,抗生素被用于人和畜禽细菌性感染疾病的治疗。然而,随着集约化养殖业的发展,抗生素被大量添加于饲料中来预防畜禽和鱼虾的养殖疾病。因此,环境介质中抗生素种类和含量随着畜禽和水产养殖业的快速发展逐年增加。本文综述了环境中抗生素的来源、残留浓度及其环境微生物生态学效应。医用、兽用抗生素和人畜粪便的农用是抗生素进入环境的主要来源,其在不同环境介质中残留浓度不一：地表水含量为μg?L-1,土壤含量为?g?kg-1,沉积物含量为μg?kg-1—mg?kg-1之间。抗生素进入土壤、水和沉积物等环境介质,经吸附-解吸、迁移和降解等过程重新分配,其降解方式主要有水解、光解和生物降解。抗生素影响环境介质中微生物的生物量、活性和群落结构,并诱导产生抗性基因,但对生态系统服务及其功能的干扰和影响尚有待进一步研究。     

微生物法检测实验动物血清中的抗生素残留

目的建立实验动物血清中抗生素残留检测方法,保证微生物质量检测结果的准确。方法采用微生物抑制法,用嗜热脂肪芽孢杆菌作为指示菌,对1640份实验动物血清进行抗生素残留检测。并与Premi@Test抗生素检测试剂盒进行比对。结果在受试样品中,检测出小鼠、大鼠、豚鼠、兔和犬的血清中存在抗生素残留,阳性率分别为27.2%、16.4%、39.9%、23.7%和47.6%。抽取82份血清和18份阴阳对照品,与Premi@Test检测的结果相比较差异不显著（X2=16.680,P〉0.05）。血清中抗生素的残留与DHL琼脂平皿的菌落生长有一定相关性（X2=9.939,P〈0.05）。结论本研究采用嗜热脂肪芽孢杆菌作为指示菌的微生物抑制法,能够对动物血清中的抗生素进行检测,具有良好的敏感性和重复性,成本低,操作简便。为实验动物体内抗生素残留检测和微生物学检测结果的准确性提供依据。     

昆虫肠道微生物及其功能研究方法进展

昆虫是世界上种类最为丰富、分布最为广泛的动物类群，其肠道内栖息着复杂且多样的微生物。不同昆虫因肠道结构、肠道内环境、食性、龄期、外界环境不同，肠道内微生物组成与丰度也存在差异。肠道微生物主要通过垂直方向与水平方向在种群与个体间传播，对昆虫宿主营养代谢、生理行为、防御、解毒等诸多方面有重要影响；通过体外培养的方法可从培养基对昆虫肠道微生物进行分离，用基质辅助激光解析电离飞行时间质谱(matrix-assisted laser desorption ionization-time of flight mass spectrometry, MALDI-TOF MS)及16S rRNA基因测序技术等可迅速鉴定微生物；宏基因组学、蛋白质组学、代谢组学等多种组学技术联合运用，使得肠道微生物鉴定与功能推测更为高效；体外试验、微生物补充、菌群移植、沉默微生物成员相关基因等试验方法使微生物功能验证更为准确；利用高温处理、溶菌酶处理、无菌饲养处理及抗生素处理等方式能清除肠道内的微生物群落，获得无菌昆虫用于功能验证试验，但当前使用最广的抗生素法在实际应用中仍存在一定局限；利用肠道微生物特性，通过共生菌基因工...     

蝇蛆肠道微生物菌群对猪粪残留抗生素的降解及抗性基因的影响

【目的】过度使用抗生素作为动物饲料添加剂,导致畜禽粪便已成为抗生素抗性基因的主要蓄积库,为了研究蝇蛆(Musca domestica)对猪粪中残留抗生素及抗性基因的影响,本文动态采集了实际农场条件下蝇蛆转化过程中猪粪堆体及虫体样本。【方法】利用q PCR、液相色谱-电喷雾质谱、同位素内标法、Illumina高通量测序以及局部相似性研究蝇蛆生物转化过程中残留抗生素降解效能及相关抗性基因组变化的微生物生态机制。【结果】6 d周期内,猪粪中四环素、土霉素、金霉素、强力霉素、磺胺嘧啶、诺氟沙星、氧氟沙星、环丙沙星以及恩诺沙星等9种残留抗生素含量显著下降,累积减量为34.3%–58.1%,每日减量百分比介于7.8%–57.4%之间。猪粪中共检测到的158种抗性基因,其中有118种大幅衰减,衰减量平均达79.3%;23种抗性基因存在富集现象,富集倍数平均为3.48。在蝇蛆肠道的作用下,粪源微生物群落中Bacteroidetes相对丰度下降,Proteobacteria相对丰度增加,尤其是Ignatzschineria增幅最大。网络分析发现,抗性基因的增减与微生物群落的变化显著相关,与抗性基因衰减相关的微生物主要属于Clostridiales和Bacteroidales,而与抗性基因富集相关的微生物主要为Alcaligenaceae、[Weeksellaceae]及Bacillales。【结论】蝇蛆可有效削减猪粪中的残留抗生素及防控抗性基因向环境扩散。     

生物泥浆反应器中多环芳烃微生物降解调控因子研究

多环芳烃 (PAHs)是一类普遍存在的环境中的优先有机污染物 ,在土壤中残留持久并难以降解 ,因此给环境和人体健康带来潜在危胁 [5] 。研究表明 ,微生物降解是去除 PAHs的主要途径 [6]。微生物能够以利用 PAHs作为碳源与能源和共代谢两种方式降解 PAHs[7] 。泥浆反应器作为生物修复技术之一 ,具有处理周期短 ,降解条件易于控制和处理效果好的特点[8,9] ,目前已成为一种重要的处理技术 ,在欧、美等发达国家 ,受到广泛重视。Grosser[10 ]等人发现 ,把微生物从污染地区分离培养后 ,接种到污染土壤中可大大提高芘的降解率。泥浆反应器中污染…     

Resistance to antibacterial drugs of Enterobacteriaceae isolated from healthy children and from those with Salmonella infections

A total of 2329 Enterobacteriaceae strains in bacterial associations isolated from healthy children and children with salmonellosis were tested for their resistance to antimicrobial drugs. It was shown that the aerobic microbial associations isolated from the healthy children contained higher numbers of strains sensitive or resistant to 1-3 antibiotics while the microbial associations from the children patients with salmonellosis treated with antibiotics contained higher numbers of strains resistant to 6-8 antibiotics. Resistance of the aerobic bacterial associations was mainly defined by resistance of E. coli and K. pneumoniae. The feces of the healthy children never treated with antimicrobial drugs contained strains resistant to them. The use of the antibiotics in the treatment led to increasing numbers of the resistant bacteria and changing species composition of the bacterial associations.     

Status of pathogens,antibiotic resistance genes and antibiotic residues in wastewater treatment systems

Pathogens are becoming nearly untreatable due to the rise in gaining new resistance against standard antibiotics. Coexistence of microbial pathogens, antibiotics and antibiotic resistant genes (ARGs) in wastewater treatment plants (WWTP) provide favourable conditions for the development of new antibiotic resistant bacteria (ARB); facilitate horizontal gene transfer among pathogens and may also serve as a hotspot for the spread of ARB and genes into the environment. In this study, the current status of wastewater treatment systems in the removal of pathogens, ARGs, and antibiotic residues are discussed. WWTP are efficient in removing pathogens and antibiotic residues to a greater extend during secondary and tertiary treatment processes. Recent studies, however, have shown high variations in the presence of pathogens including ARB as well as antibiotic resistance genes (ARG) in the final effluent. Prolonged sludge retention time (SRT) and hydraulic retention time (HRT) during secondary treatment will facilitate antibiotic removal by adsorption and biodegradation. However, the above conditions can also lead to the enhancement of antibiotic resistance process in microbes. Therefore, optimum conditions for the operation of conventional WWTP for the efficient removal of antibiotics are yet to be established. The removal of antibiotic residues can be accelerated by combining conventional activated sludge (CAS) process with an additional treatment technology involving dosing with ozone. The advanced biological treatment method using membrane bioreactors (MBR) in combination with coagulation reportedly has the best ARG removal efficiency, and removes both ARB and extracellular ARGs. While studies have predicted the fate for ARGs in wastewater treatment plants, the mechanisms of ARGs acquisition remains to be conclusively established. Thus, strategies to investigate the underlying mechanism of acquisition of ARGs within the WWTP are also provided in this review.     

抗生素产生菌诱变育种的研究进展

一种抗生素能否产业化决定于抗生素产生菌菌种的产素水平。因而如何提高抗生素产生菌的产素水平成为抗生素研究工作者长期不懈探索的重要课题。其中,诱变育种以其技术设备简单、省时省力等优点得到了微生物育种工作者的青睐,在诱变育种技术上取得了巨大进展,为抗生素工业化生产发挥了重要作用。综述了抗生素产生菌的诱变育种研究进展。     

Response of nursery pigs to a synbiotic preparation of starch and an anti-Escherichia coli K88 probiotic

Postweaning diarrhea in pigs is frequently caused by enterotoxigenic Escherichia coli K88 (ETEC). The aim of this study was to test the efficacy of E. coli probiotics (PRO) in young pigs challenged with E. coli K88. We also tested the synbiotic interaction with raw potato starch (RPS), which can be used as a prebiotic. Forty 17-day-old weaned piglets were randomly assigned to four treatments: treatment 1, positive-control diet (C), no probiotics or RPS but containing in-feed antibiotics; treatment 2, probiotic (PRO), no feed antibiotics plus a 50:50 mixture of probiotic E. coli strains UM-2 and UM-7; treatment 3, 14% RPS, no antibiotics (RPS); treatment 4, 14% RPS plus a 50:50 mixture of probiotic E. coli strains UM-2 and UM-7, no antibiotics (PRO-RPS). The pigs were challenged with pathogenic E. coli K88 strains on day 7 of the experiment (24-day-old pigs) and euthanized on day 10 of the experiment (35-day-old pigs). Probiotic and pathogenic E. coli strains were enumerated by selective enrichment on antibiotics, and microbial community analysis was conducted using terminal restriction length polymorphism analysis (T-RFLP) of 16S rRNA genes. The combination of raw potato starch and the probiotic had a beneficial effect on piglet growth performance and resulted in a reduction of diarrhea and increased microbial diversity in the gut. We conclude that the use of E. coli probiotic strains against E. coli K88 in the presence of raw potato starch is effective in reducing the negative effects of ETEC in a piglet challenge model.     

Coselection for microbial resistance to metals and antibiotics in freshwater microcosms

Bacterial resistances to diverse metals and antibiotics are often genetically linked, suggesting that exposure to toxic metals may select for strains resistant to antibiotics and vice versa. To test the hypothesis that resistances to metals and antibiotics are coselected for in environmental microbial assemblages, we investigated the frequency of diverse resistances in freshwater microcosms amended with Cd, Ni, ampicillin or tetracycline. We found that all four toxicants significantly increased the frequency of bacterioplankton resistance to multiple, chemically unrelated metals and antibiotics. An ampicillin-resistant strain of the opportunistic human pathogen Ralstonia mannitolilytica was enriched in microcosms amended with Cd. Frequencies of antibiotic resistance were elevated in microcosms with metal concentrations representative of industry and mining-impacted environments (0.01-1 mM). Metal but not antibiotic amendments decreased microbial diversity, and a weeklong exposure to high concentrations of ampicillin (0.01-10 mg l-1) and tetracycline (0.03-30 mg l-1) decreased microbial abundance only slightly, implying a large reservoir of antibiotic resistance in the studied environment. Our results provide first experimental evidence that the exposure of freshwater environments to individual metals and antibiotics selects for multiresistant microorganisms, including opportunistic human pathogens.     

Do we need new antibiotics? The search for new targets and new compounds

Resistance to antibiotics and other antimicrobial compounds continues to increase. There are several possibilities for protection against pathogenic microorganisms, for instance, preparation of new vaccines against resistant bacterial strains, use of specific bacteriophages, and searching for new antibiotics. The antibiotic search includes: (1) looking for new antibiotics from nontraditional or less traditional sources, (2) sequencing microbial genomes with the aim of finding genes specifying biosynthesis of antibiotics, (3) analyzing DNA from the environment (metagenomics), (4) reexamining forgotten natural compounds and products of their transformations, and (5) investigating new antibiotic targets in pathogenic bacteria.     

微生物源农用抗生素的研发与高产策略

开发高效、低毒、低残留的绿色农药是农药研发的发展趋势,其中微生物源农药抗生素占据了重要地位。随着基因组学、代谢工程、高通量筛选等技术的发展,新型微生物源农用抗生素的研究进入了新的阶段。文中简要总结了近10年来研发的新型微生物源农用抗生素的种类、农用抗生素产生菌株的高产育种与发酵研究策略等,为未来农用抗生素的研发提供参考。     

海口市污水处理厂活性污泥中细菌的分离鉴定及耐药性分析

为了解海口市白沙门污水处理厂活性污泥中细菌抗生素耐性情况,采用平板分离技术分离、纯化细菌,并通过BIOLOG微生物鉴定系统对筛选到的细菌进行鉴定,同时采用Kirby-Bauer纸片琼脂扩散法进行药敏试验并进行抗生素耐性分析。本研究共分离到18株细菌,分属8个属,14个种,其中G+和G-均为9株。抗生素药敏性试验结果表明,所有菌株均耐药,菌株单重耐药率、双重耐药性及多重耐药性分别为50%、38.9%、和11.1%。菌株对9种常用抗生素:头孢他啶、环丙沙星、庆大霉素、链霉素、氨苄西林、红霉素、氯霉素、四环素、卡那霉素的耐药率分别为61.1%、0%、5.6%、16.7%、50%、16.7%、11.1%、0%、5.6%。综上所述,白沙门污水处理厂活性污泥中的细菌耐药性比较严重,存在潜在的环境生态和人畜健康风险。本研究揭示了当前白沙门污水处理厂活性污泥中细菌对常见抗生素耐药的严重现状,为建议污水处理厂加强出水及污泥中抗生素耐药性及耐药基因的检测并评估其生态影响提供基础,避免出水及污泥中的抗性菌和耐药基因可能带来的风险问题。     

群体感应在生物强化功能菌定殖及降解能力增强中的作用研究进展

由于难降解有机污染物和外界环境对水处理系统的冲击干扰,污水水质常出现不达标现象。引入外源含有相关功能基因并且具有基因水平转移能力的工程菌株进行生物强化处理是提高污水处理效能的有效措施。污水处理系统中存在能够分泌信号分子的菌体,菌间具有群体感应现象,当种群密度达到感应阈值时,菌体会通过释放信号分子来触发一些群体行为,从而激活相关基因的表达(如生物膜形成、生物发光、抗生素合成和毒力因子表达等)。早期的群体感应技术研究主要集中在信号传递学、微生物社会行为学和医学微生物领域,近年来,在水处理领域也开始有相继报道,研究表明群体感应在污水生物处理中发挥重要作用,并且影响生物强化菌株的定殖和污染物降解,因此群体感应行为调控是生物强化技术成效显著与否的关键因素。本文综述了群体感应及信号分子的作用机制、信号分子释放及存在的影响因素以及群体感应对菌株定殖、微生物群落结构和污染物去除的影响,并对从群体感应角度出发研究生物强化过程进行了展望,旨在为生物强化技术的有效实施及提升污水处理效能提供一种新思路,为深入理解生物强化过程中群体感应调控行为提供理论参考。