耐药菌:难以对付的敌人

40年代、青霉索作为第一种应用于临床的抗生素,成功解决了临床上金葡菌感染这一难题。随后问世的大环内酯类,氨基甙类抗生素又使肺炎、肺结核的死亡率降低了80％,当时曾有人断言:人类战胜细菌的时代已经到来。但是,事实并不像人们想像的那样美好,许多抗生素在应用多年后出现了不程度的药效减低,天然青霉素在控制金葡菌感染方面几乎已失去药用价值。医学家在研究这一现象后惊讶地发现,在和抗生素接触多次后,细菌已世化出一整套有效的耐药机制,耐药菌这个隐藏的敌人正在逐渐强大起来。     

紫外消毒对城市污水处理厂耐药菌去除的影响研究

通过选取假单胞杆菌、鲍曼不动杆菌以及芽孢杆菌3种耐药菌作为研究对象,研究紫外光变化对这3种耐药菌的去除效果,得出较好的紫外消毒条件,应用到实际的污水处理厂消毒过程中。结果表明,紫外消毒对3种耐药菌都有很好的灭活效果。     

养殖鱼塘底泥微生物抗生素耐药基因分布分析

针对水产养殖中抗生素滥用引起微生物耐药基因污染的现象,本实验以杭州市某水产养殖区底泥样品为研究对象,测定了底泥样品中抗生素含量,并从其中分离出74株可培养细菌.采用PCR方法对分离菌株中磺胺类、四环素类、喹诺酮类、氯霉素类抗性基因(antibiotic resistance genes,ARGs)和整合子基因进行了检测.结果表明,除tet A(四环素类)耐药基因未被检出外,其余3种耐药基因及整合子基因均被检出,其中qnr S(喹诺酮类)耐药基因的检出率最高,为50.00%.另外,利用16S r DNA序列分析技术将分离菌株鉴定为12个属、19个种,包括环境中多种常见土著细菌及部分致病菌,其中气单胞菌(Aeromonas spp.)数量最多(29株),占分离菌株的39.19%.掌握水产养殖区中ARGs的污染现状,对控制其传播、保障食品安全和保护微生态环境具有重要指导意义.     

气溶胶中抗生素抗性基因研究进展:以养殖场和医院为例

抗生素在医药和养殖业的大量使用导致耐药菌的出现,加速了抗生素抗性基因(antibiotic resistance genes,ARGs)在不同环境介质中的传播扩散,抗生素耐药性已成为目前全球卫生、食品安全和发展的最大威胁之一。气溶胶作为ARGs的潜在储存库缺乏系统的研究数据,而通过空气传播具有较高抗生素抗性水平的细菌可能是引起重要疾病的主要传播途径。本文针对养殖场和医院2个抗生素大量使用的典型场所,对气溶胶中ARGs的污染现状、样品采集与检测技术进行综述,并探讨了这一环境污染的潜在风险,表明开展气溶胶中ARGs研究的必要性,并为以后需开展的工作提出几点建议。     

连续施用发酵猪粪对土壤中四环素抗性基因数量的影响

为了研究连续施用发酵猪粪之后,土壤中抗性基因的数量以及发酵猪粪的施用量对土壤抗性基因的影响,采用实时荧光定量PCR方法,对稻麦轮作模式下连续6年施用4.5t/hm2和9.0t/hm2发酵猪粪的土壤进行了四环素抗性基因(TRGs)的检测和定量分析.在施用发酵猪粪的土壤中能够检测到9种TRGs.施用发酵猪粪显著增加了土壤中tet G、tet L、tet B(P)、tet O、tet W的绝对数量,其中tet B(P)、tet W、tet O的绝对数量受到了施肥用量的影响;而tet Z、tet C和tet S的绝对数量则没有受到施肥的影响.上述8种TRGs在同一施肥处理的0~5cm、5~10cm以及10~20cm土壤中的相对丰度分布均无显著差异.发酵猪粪的施用也显著增加了tet G、tet L、tet B(P)和tet O的相对丰度,但是仅有tet O的相对丰度受到了施肥用量的显著影响.本研究表明,在稻麦轮作模式下,农田土壤中的TRGs数量会受到发酵猪粪中残留TRGs的影响,连续施用该粪肥会显著增加土壤中tet G、tet L、tet B(P)和tet O的绝对数量和相对丰度.因此,需要优化畜禽粪堆肥发酵工艺,以减少抗性基因在粪肥中的残留.     

混合抗生素对大肠杆菌耐药性的联合诱导效应

群体感应抑制剂（quorum sensing inhibitors,QSIs）作为一类新型抗生素,具有广阔的应用前景,目前已被使用在水产养殖、污水治理和农作物病害防治等领域中,其与传统抗生素极有可能在环境中共存。因此,有必要探究QSIs与传统抗生素对细菌耐药性的联合效应。以往的研究往往采用单位法探究固定浓度下混合抗生素的联合耐药效应,但单位法无法在较大浓度范围评价其联合耐药风险。该研究以RP4质粒接合转移表征细菌耐药性的传播,将独立作用模型（independent action,IA）应用到联合耐药作用的判别中,以喹诺酮（quinolones,QNs）作为传统抗生素的代表,探究了QSIs与QNs对大肠杆菌（Escherichia coli）RP4质粒接合转移的联合效应。结果表明,15组QSIs-QNs混合物均能诱导E. coli产生接合转移效应,混合物的最大促进率在23.32%～73.07%。对比IA曲线与混合物对E. coli接合转移的剂量-效应曲线的位置关系,发现QSIs-QNs对E. coli的联合接合作用均表现出浓度依赖特征,即混合物促进E. coli接合转移频率时的联合作用模...     

水环境中耐热大肠菌群的抗生素耐药性与质粒谱研究

用滤膜法、mFC培养基从5种水体中分离出疑似耐热大肠菌162株,用API微生物分析系统鉴定到种,以Kirby-Bauer法分析其对人畜常用10种抗生素的耐药性,碱裂解法小量制备各菌株质粒DNA做质粒谱分析.结果表明,埃希氏大肠杆菌为优势菌,占分离菌总数的96.3%.除分离自泉水的3株外,其它菌株都对3种及3种以上抗生素耐药,多重耐药率为98.1%.不同水体分离菌株对氧氟沙星、诺氟沙星、庆大霉素、丁胺卡那霉素、链霉素的耐药率有显著性差异(P<0.005).92株菌(56.8%)提取到大小为0.90～158.83kb、数量为1～6个的质粒,有81种质粒谱型.70株(43.2%)未提取到质粒的细菌中有67株为多重耐药.具有相同质粒谱型的菌株耐药谱都不相同.未发现质粒谱与抗生素耐药性间有明显相关性.图1表3参15     

Identification and characterization of integron mediated antibiotic resistance in pentachlorophenol degrading bacterium isolated from the chemostat

A bacterial consortium was developed by continuous enrichment of microbial population isolated from sediment core of pulp and paper mill e uent in mineral salts medium (MSM) supplemented with pentachlorophenol (PCP) as sole source of carbon and energy in the chemostat. The consortia contained three bacterial strains. They were identified as Escherichia coli, Pseudomonas aeruginosa and Acinetobacter sp. by 16S rRNA gene sequence analysis. Acinetobacter sp. readily degraded PCP through the formation of tetrachloro-phydroquinone (TecH), 2-chloro-1,4-benzenediol and products of ortho ring cleavage detected by gas chromatograph/mass spectrometer (GC-MS). Out of the three acclimated PCP degrading bacterial strains only one strain, Acinetobacter sp. showed the presence of integron gene cassette as a marker of its stability and antibiotic resistance. The strain possessed a 4.17 kb amplicon with 22 ORF’s. The plasmid isolated from the Acinetobacter sp. was subjected to shotgun cloning through restriction digestion by BamHI, HindIII and SalI, ligated to pUC19 vector and transformed into E. coli XLBlue1 , and finally selected on MSM containing PCP as sole source of carbon and energy with ampicillin as antibiotic marker. DNA sequence analysis of recombinant clones indicated homology with integron gene cassette and multiple antibiotic resistance genes.     

Screening and characterization of extremophiles in spacecraft assembly environment北大核心CSCD

Spacecrafts need to strictly control microorganisms before entering space flight. The Spacecraft Assembly, Integration, and Test Center (AITC) is an important environmental source for spacecrafts to carry microorganisms. The assembly clean room has characteristics of ventilation, dryness, and lack of nutrients that are not conducive to the growth and reproduction of microorganisms, except for extremophiles. In this study, based on the internal air and surface environment of the AITC in China, 13 strains of bacteria were identified by plate culture and 16S sequence analysis, and their extreme environmental tolerance, antibiotic resistance, and film-forming ability were tested. All these bacteria belonged to the Firmicutes and Proteobacteria phyla, and nine strains belonged to the Bacillus genus. All 13 strains of bacteria were salt-tolerant, acid-tolerant, and alkali-tolerant, and 69.2% of the bacteria survived heat shock at 80 ℃. Among these, one strain of Sphingomonas sp. JCM7513 isolated from the surface environment was insensitive to all the tested antibiotics and had strong drug resistance, while the other 12 strains were sensitive to most β-lactam antibiotics but had strong resistance to tetracyclic antibiotics and erythromycin. Most of the isolated bacteria exhibited strong biofilm-forming abilities. The study showed that there are a certain number of extremophiles in the spacecraft assembly environment. To protect spacecrafts from biological corrosion and planetary protection forward pollution, more effective monitoring and elimination methods are needed. © 2022 Science Press. All rights reserved.     

红谷霉素对细菌耐药性质粒转移的抑制作用

以Shigella flexneri D15 R-plasmid CmR和Escherichia coil 1485 RifR为试验菌株,研究红谷霉素对细菌耐药性质粒在细菌之间转移的抑制作用及对两种细菌R质粒的消除作用.将红谷霉素(30μg/mL)与S.flexneri D15 R-plasmid CmR和E.coli 1485 RifR按实验设计混合并分别培养24 h和48 h,涂布在麦康凯琼脂平板上,计算相应菌落数.结果表明,当红谷霉素抑菌浓度为30μg/mL时,对细菌耐氯霉素的R质粒的转移抑制率分别达到92.82%(24 h培养)和96.04%(48 h培养),但对R质粒的消除作用不显著.说明红谷霉素对R质粒转移抑制作用强烈但不具有R质粒的消除作用.