食源性乳酸菌抗药性分析

抗生素的不合理使用导致病原菌耐药性已成为威胁人类健康的公共问题。乳酸菌在食品发酵、饲料青贮等领域具有悠久的应用历史,是公认的安全性菌株。但是研究发现,有些乳酸菌具有抗药性,根据抗药因子的可转移性,乳酸菌的抗药性可分为固有抗药性和外获抗药性。外获抗药性多由接合质粒或转座因子编码,这些抗药基因随发酵食品或益生菌剂进入肠道,是否与肠道共生菌或病原菌进行传递进而对机体带来危害引起了人们的关注。本文就食源性乳酸菌抗药性产生的分子机制、检测方法以及可能的抗药基因水平转移方式进行综述,以便对食源性乳酸菌的抗药性有一个全面认识。     

Emergence of vancomycin tolerance in Streptococcus pneumoniae.

Streptococcus pneumoniae, the pneumococcus, is the most common cause of sepsis and meningitis. Multiple-antibiotic-resistant strains are widespread, and vancomycin is the antibiotic of last resort. Emergence of vancomycin resistance in this community-acquired bacterium would be catastrophic. Antibiotic tolerance, the ability of bacteria to survive but not grow in the presence of antibiotics, is a precursor phenotype to resistance. Here we show that loss of function of the VncS histidine kinase of a two-component sensor-regulator system in S. pneumoniae produced tolerance to vancomycin and other classes of antibiotic. Bacterial two-component systems monitor environmental parameters through a sensor histidine-kinase/phosphatase, which phosphorylates/dephosphorylates a response regulator that in turn mediates changes in gene expression. These results indicate that signal transduction is critical for the bactericidal activity of antibiotics. Experimental meningitis caused by the vncS mutant failed to respond to vancomycin. Clinical isolates tolerant to vancomycin were identified and DNA sequencing revealed nucleotide alterations in vncS. We conclude that broad antibiotic tolerance of S. pneumoniae has emerged in the community by a molecular mechanism that eliminates sensitivity to the current cornerstone of therapy, vancomycin.     

Plasmids of the same Inc groups in Enterobacteria before and after the medical use of antibiotics

Conjugative plasmids were common in enterobacteria isolated before the medical use of antibiotics. Plasmid F of Escherichia coli K-12 was one example and we identified others in over 20% of a collection of strains isolated between 1917 and 1954, the Murray collection. In the past 25 years, conjugative plasmids encoding antibiotic resistances have become common in bacteria of the same genera as those of the Murray Collection--Salmonella, Shigella, Klebsiella, Proteus, Escherichia. The present study was made to show whether the 'pre-antibiotic' plasmids belonged to the same groups, as defined by incompatibility tests (Inc groups), as modern R plasmids. Of 84 such plasmids established in E. coli K-12, none with antibiotic resistance determinants, 65 belonged to the same groups as present resistance (R) plasmids. Thus the remarkable way in which medically important bacteria have acquired antibiotic resistance in the past 25 years seems to have been by the insertion of new genes into existing plasmids rather than by the spread of previously rare plasmids.     

抗生素与肠道菌群关系研究进展

 从抗生素与肠道菌群的关系出发,在抗生素角度,概述了抗生素对肠道菌群构成、肠道定植抗力、菌群类别及代谢活动的影响,综述了抗生素通过调节肠道菌群恢复肠-脑轴、肠-肝轴稳态,延缓非感染性疾病的研究进展。在肠道菌群角度,简述了肠道菌群在抗生素耐药基因储存传播、新型抗生素发现过程中的特别作用。     

Vancomycin-resistant enterococci exploit antibiotic-induced innate immune deficits

Infection with antibiotic-resistant bacteria, such as vancomycin-resistant Enterococcus (VRE), is a dangerous and costly complication of broad-spectrum antibiotic therapy. How antibiotic-mediated elimination of commensal bacteria promotes infection by antibiotic-resistant bacteria is a fertile area for speculation with few defined mechanisms. Here we demonstrate that antibiotic treatment of mice notably downregulates intestinal expression of RegIIIgamma (also known as Reg3g), a secreted C-type lectin that kills Gram-positive bacteria, including VRE. Downregulation of RegIIIgamma markedly decreases in vivo killing of VRE in the intestine of antibiotic-treated mice. Stimulation of intestinal Toll-like receptor 4 by oral administration of lipopolysaccharide re-induces RegIIIgamma, thereby boosting innate immune resistance of antibiotic-treated mice against VRE. Compromised mucosal innate immune defence, as induced by broad-spectrum antibiotic therapy, can be corrected by selectively stimulating mucosal epithelial Toll-like receptors, providing a potential therapeutic approach to reduce colonization and infection by antibiotic-resistant microbes.     

儿科急性呼吸道感染滥用抗生素现状的分析与对策

目的探讨儿科医生对治疗急性呼吸道感染滥用抗生素的情况。方法通过查阅有关文献报道和近几年来所了解到情况,对儿科治疗急性呼吸道感染滥用抗生素的现状、引起原因、后果及防止策略作一综述。结果儿科急性呼吸道感染抗生素使用频率最高,滥用现象十分普遍、危害极为严重,其原因复杂,控制困难。结论通过共同努力如能使儿童呼吸道感染步入合理使用抗生素的轨迹,对减少细菌耐药,减少抗生素不良反应,节约有限的医疗资源等均有重大的现实意义。     

SOS response promotes horizontal dissemination of antibiotic resistance genes

Mobile genetic elements have a crucial role in spreading antibiotic resistance genes among bacterial populations. Environmental and genetic factors that regulate conjugative transfer of antibiotic resistance genes in bacterial populations are largely unknown. Integrating conjugative elements (ICEs) are a diverse group of mobile elements that are transferred by means of cell-cell contact and integrate into the chromosome of the new host. SXT is a approximately 100-kilobase ICE derived from Vibrio cholerae that encodes genes that confer resistance to chloramphenicol, sulphamethoxazole, trimethoprim and streptomycin. SXT-related elements were not detected in V. cholerae before 1993 but are now present in almost all clinical V. cholerae isolates from Asia. ICEs related to SXT are also present in several other bacterial species and encode a variety of antibiotic and heavy metal resistance genes. Here we show that SetR, an SXT encoded repressor, represses the expression of activators of SXT transfer. The 'SOS response' to DNA damage alleviates this repression, increasing the expression of genes necessary for SXT transfer and hence the frequency of transfer. SOS is induced by a variety of environmental factors and antibiotics, for example ciprofloxacin, and we show that ciprofloxacin induces SXT transfer as well. Thus, we present a mechanism by which therapeutic agents can promote the spread of antibiotic resistance genes.     

肠球菌临床耐药性分析及耐药基因vanM在屎肠球菌中的检测

目的了解肠球菌临床分离株耐药性及vanM基因在屎肠球菌的分布情况,为临床合理用药及探讨耐药机制提供依据.方法采用VITEK60全自动微生物分析仪对肠球菌进行鉴定及药敏试验,按NCCLS/CLSI标准判断并统计分析结果;PCR法检测耐万古霉素、替考拉宁菌株中的耐药基因vanM,并对阳性基因进行测序,分析耐药基因与耐药性的关系.结果临床分离肠球菌共506株,其中屎肠球菌277株,粪肠球菌229株.尿液标本来源比例最高,为167株（33%）;粪便标本114株（22.5%）;痰液标本78株（15.4%）;胆汁标本57株（11.3%）;脓液标本54株（10.7%）;其他来源36株（7.1%）.药敏结果显示：肠球菌对大部分临床抗菌药高度耐药,屎肠球菌对β-内酰胺类抗菌药物（氨苄西林）、喹诺酮类（左氧氟沙星）、糖肽类（万古霉素、替考拉宁）的耐药率明显高于粪肠球菌（P〈0.05）.vanM基因在耐万古霉素菌株中的检出率为100%,耐替考拉宁菌株检出率为90%.结论屎肠球菌可引起临床各类感染,屎肠球菌对大多数抗生素的耐药率明显高于粪肠球菌,且呈多重耐药趋势.vanM基因可能在糖肽类药物万古霉素、替考拉宁耐药机制中发挥重要作用.     

Gene expression in Pseudomonas aeruginosa biofilms.

Bacteria often adopt a sessile biofilm lifestyle that is resistant to antimicrobial treatment. Opportunistic pathogenic bacteria like Pseudomonas aeruginosa can develop persistent infections. To gain insights into the differences between free-living P. aeruginosa cells and those in biofilms, and into the mechanisms underlying the resistance of biofilms to antibiotics, we used DNA microarrays. Here we show that, despite the striking differences in lifestyles, only about 1% of genes showed differential expression in the two growth modes; about 0.5% of genes were activated and about 0.5% were repressed in biofilms. Some of the regulated genes are known to affect antibiotic sensitivity of free-living P. aeruginosa. Exposure of biofilms to high levels of the antibiotic tobramycin caused differential expression of 20 genes. We propose that this response is critical for the development of biofilm resistance to tobramycin. Our results show that gene expression in biofilm cells is similar to that in free-living cells but there are a small number of significant differences. Our identification of biofilm-regulated genes points to mechanisms of biofilm resistance to antibiotics.     

水环境中抗生素和抗性基因污染特征及控制措施

 抗生素作为一类抗菌类药物被广泛用于医疗、农业和畜牧业等领域，因其使用量大并能诱导产生耐药菌株，对人类健康和生态环境造成巨大威胁。在梳理近年来地表水环境中抗生素相关研究的基础上，阐述了水环境中抗生素和抗性基因的污染来源和污染特征，分析了环境浓度水平下抗生素污染对人群和生态环境的影响，讨论了水环境中抗生素污染的控制措施及目前研究的主要问题，并对今后的研究进行了展望。     

大肠埃希菌ESBLs检测及其耐药性分析

目的探讨本地区大肠埃希菌分离株中超广谱β-内酰胺酶（ESBLs）的阳性率及产酶株对β-内酰胺类抗生素的耐药性。方法采用双纸片协同法检测菌株的ESBLs,并用Kirby-Bauer法检测了415株产ESBLs的大肠埃希菌对16种β-内酰胺类抗生素的耐药性,并对其标本来源和病区分布进行了分析。结果ESBLs总检出率为50．8％,其中ESBLs检出率较高的科室为普外科、脑外科、肾内科、骨科、呼吸内科。所有大肠埃希菌分离株均对Imipenem和Meropenem敏感。产ESBLs株对其他14种β-内酰胺类抗生素的耐药性均高于非产酶株。结论本地区ESBLs检出率较高,分布科室有特殊性,产ESBLs株对抗生素的耐药性高于非产酶株。     

A genetic basis for Pseudomonas aeruginosa biofilm antibiotic resistance

Biofilms are surface-attached microbial communities with characteristic architecture and phenotypic and biochemical properties distinct from their free-swimming, planktonic counterparts. One of the best-known of these biofilm-specific properties is the development of antibiotic resistance that can be up to 1,000-fold greater than planktonic cells. We report a genetic determinant of this high-level resistance in the Gram-negative opportunistic pathogen, Pseudomonas aeruginosa. We have identified a mutant of P. aeruginosa that, while still capable of forming biofilms with the characteristic P. aeruginosa architecture, does not develop high-level biofilm-specific resistance to three different classes of antibiotics. The locus identified in our screen, ndvB, is required for the synthesis of periplasmic glucans. Our discovery that these periplasmic glucans interact physically with tobramycin suggests that these glucose polymers may prevent antibiotics from reaching their sites of action by sequestering these antimicrobial agents in the periplasm. Our results indicate that biofilms themselves are not simply a diffusion barrier to these antibiotics, but rather that bacteria within these microbial communities employ distinct mechanisms to resist the action of antimicrobial agents.     

尿路感染病原菌及耐药性分析

目的：探讨尿路感染的病原菌构成及耐药性,指导临床合理使用抗菌药物。方法：收集我院2006年1月至2007年12月尿路感染患者清洁中段尿培养阳性588株细菌进行鉴定,并用K—B纸片法作药敏分析。结果：尿路感染病原菌以革兰阴性杆菌为主（63．8％）,前4位分别是大肠埃希菌（45．7％）、凝固酶阴性葡萄球菌（11.6％）、假丝酵母菌（11．1％）、肠球菌（7．8％）。大肠埃希菌和克雷伯菌超广谱β-内酰胺胺酶（ESBLS）检出率分别为29％和32.1％,甲氧西林耐药凝固酶阴性葡萄球菌（MRCNS）检出率为41．2％。结论：革兰阴性杆菌是尿路感染的主要病原菌,对常规抗菌药物的耐药性呈上升趋势,细菌分离培养鉴定及药敏试验对指导临床合理使用抗菌药物具有重要意义。     

DNA cloning in Streptomyces: resistance genes from antibiotic-producing species

The biochemical and morphological differentiation of actinomycetes makes them academically and economically interesting. Their secondary metabolites provide the majority of medically and agriculturally important antibiotics (streptomycete genes may also be the primary source of clinically important antibiotic resistance); their complex morphological developmental cycle involves a series of changes from vegetative mycelial growth to spore formation. Recombinant DNA technology would add a powerful new dimension to the analysis of these various aspects of actinomycete biology and would also facilitate the development of industrial strains with increased antibiotic yield, or capable of making new antibiotics. For most of these purposes, cloning of genes within and between actinomycetes is required to study the expression of particular genes in genetic backgrounds defined by mutations of the characters under study. To achieve this, we have now developed a method for molecular cloning involving the transfer of genes between unrelated streptomycetes.     

Cu2+对UV-nTiO2光催化降解抗生素的影响研究

Cu~(2+)是环境中常见二价金属离子,但其对光催化降解抗生素废水的影响有待探明。为此,构建UV-nTiO_2体系并探究Cu~(2+)共存下三类抗生素(磺胺类抗生素、氯霉素类抗生素及四环素类抗生素)的光催化降解特性。结果表明,Cu~(2+)可显著影响抗生素光催化降解且呈"低浓度促进,高浓度抑制"效应。当Cu~(2+)浓度介于0.001～0.03mol/L时,抗生素降解半衰期(T_(0.5))可降低15%～90%;当浓度介于0.01～0.1mol/L时,T_(0.5)可增大1.5～4倍。结果也表明,Cu~(2+)浓度和T_(0.5)之间的关系可通过正弦函数进行表征(R~20.905),且Cu~(2+)对抗生素光催化降解效率的影响和抗生素的结构特征显著相关(P 0.05)。     

宿主体内质粒有无微生物竞争模型的种群动力学和抗生素耐药分析

研究宿主体内带质粒微生物和不带质粒微生物的竞争关系．在分析相应数学模型动力学行为的基础上，获得了依赖于具有生物学意义参数的所有8种可能的结果．在对质粒模型分析的基础上，进一步分析抗生素的杀菌效果，这些抗生素是临床上通过注射、口服和输液而进入宿主体内，结果表明抗生素的剂量越大，带质粒微生物幸存的可能性越大，也就是说产生抗生素耐受的几率越大。     

Molecular mechanisms that confer antibacterial drug resistance

Antibiotics--compounds that are literally 'against life'--are typically antibacterial drugs, interfering with some structure or process that is essential to bacterial growth or survival without harm to the eukaryotic host harbouring the infecting bacteria. We live in an era when antibiotic resistance has spread at an alarming rate and when dire predictions concerning the lack of effective antibacterial drugs occur with increasing frequency. In this context it is apposite to ask a few simple questions about these life-saving molecules. What are antibiotics? Where do they come from? How do they work? Why do they stop being effective? How do we find new antibiotics? And can we slow down the development of antibiotic-resistant superbugs?     

STEC的耐药性监测和分析

目的了解各种标本来源的产志贺样毒素大肠杆菌(STEC)对常用抗生素的敏感性.方法采用WHO推荐的K-B法,进行分鉴定菌株对18种抗生素的药物敏感试验,依照NCCLS判定结果.结果各种来源产志贺样毒素的大肠杆菌共46株,O157型23株,其余为非O157型.全部STEC对复方新诺明耐药,对链霉素耐药率为28.3%(13株),氨苄西林为30.4%(14株),红霉素为69.6%(32株),而且有5株对至少4种以上抗生素产生多重耐药,耐药谱为复方新诺明-链霉素-红霉素-氨苄西林.非O157型STEC耐药菌次为122,而O157型为63.结论非O157型STEC对抗生素可能较易形成耐药性.     

海洋链霉菌产生的抗生素S—111—9的研究

抗生素Ｓ－１１１－９是由分离自厦门地区红树根际海泥的一株暂定名为厦海链霉菌所产生的。对该菌发酵液含有的抗生素进行了提取、纯化、理化性质、紫外光谱、红外光谱、快原子轰击质谱、核磁共振谱及元素分析研究，结果表明它是一种碱性水溶性氨基环醇类抗生素，其分子量是６１４，分子式为Ｃ２７Ｈ６４Ｎ７Ｏ８。但与已知的氨基环醇类抗生素比较，在某些理论性质及波谱特征上均有不同，它是氨基环醇类抗生素的一个新成员。     

产超广谱β-内酰胺酶大肠埃希菌的临床感染分布及耐药性分析

目的-探讨医院产超广谱β-内酰胺酶（ESBLs）大肠埃希菌的临床感染分布及耐药情况,为临床抗感染治疗合理选择抗生素、减少耐药发生提供依据。方法：收集2010年8月-2011年12月从临床送检标本中分离的大肠埃希菌264株,药敏试验采用K～B纸片扩散法。根据美国临床检验室标准化委员会（CLSI）推荐的纸片扩散法,严格按照其制定的标准进行ESBLs确证实验。采用wHONET5．4软件进行统计学分析。结果：264株大肠埃希菌中检出134株（50．8％）产ESBLs大肠埃希菌,标本分布以痰标本最高（47．76％）,而科室分布以普外科最高（12．61％）。在药敏实验中,产ESBLs菌株对亚胺培南、美罗培南、头孢哌酮／舒巴坦、阿米卡星、哌拉西林／他唑巴坦、呋喃妥因、头孢西丁的耐药性较低（0％。15．5％）：环丙沙星、左氧氟沙星、庆大霉素、复方新诺明,阿莫西林／克拉维酸的耐药性较高（41.3％～82．8％）;青霉素类、头孢菌素类及ATM耐药性高（100％）,临床不宜选用。结论：产ESBLs大肠埃希菌存在多重耐药性,临床实验室应加强产ESBLs菌株的监测,根据,临床药敏结果合理选用抗生素。