The shared antibiotic resistome of soil bacteria and human pathogens

Soil microbiota represent one of the ancient evolutionary origins of antibiotic resistance and have been proposed as a reservoir of resistance genes available for exchange with clinical pathogens. Using a high-throughput functional metagenomic approach in conjunction with a pipeline for the de novo assembly of short-read sequence data from functional selections (termed PARFuMS), we provide evidence for recent exchange of antibiotic resistance genes between environmental bacteria and clinical pathogens. We describe multidrug-resistant soil bacteria containing resistance cassettes against five classes of antibiotics (β-lactams, aminoglycosides, amphenicols, sulfonamides, and tetracyclines) that have perfect nucleotide identity to genes from diverse human pathogens. This identity encompasses noncoding regions as well as multiple mobilization sequences, offering not only evidence of lateral exchange but also a mechanism by which antibiotic resistance disseminates.     

益生菌对肉仔鸭肠道结构的影响

选取1日龄樱桃谷肉鸭250羽,随机分成5组,每组5个重复,每重复10羽。对照组饲喂基础日粮,试验组饲喂基础日粮中添加0.04%益生菌替代50%抗生素试验日粮。结果表明,替代50%抗生素的地衣芽孢杆菌能显著刺激十二指肠绒毛生长,提高肠道消化能力(P<0.01);空肠结构中,绒毛高度/隐窝深度指数均显著提高(P<0.05);回肠结构中,其隐窝深度变浅,绒毛高度/隐窝深度指数显著增高(P<0.01)。结果显示,肉鸭日粮添加0.04%益生菌替代50%抗生素可提高小肠绒毛高度,降低隐窝深度,增加V/C值,改善肠道黏膜形态结构,促进肉鸭健康生产。     

Inhibition of protein synthesis by spectinomycin

Spectinomycin selectively inhibits protein synthesis in cells and in extracts of Escherichia coli. Mutations to high-level resistance to this antibiotic map close to the streptomycin locus, and the site of action of spectinomycin, like that of streptomycin, is the 30S ribosomal subunit, as shown by experiments with reconstituted 70S ribosomes containing subunits from sensitive and from resistant ribosomes. In contrast to streptomycin, however, spectinomycin is not bactericidal and causes no detectable misreading of polyribonucleotides.     

牛磺酸与抗生素复合剂对猪精液常温保存效果的影响

牛磺酸(Tau)具有良好的抗氧化性,为研究Tau与抗生素联合使用对猪精液常温保存效果的影响,以不含抗生素Modena稀释液为对照基础液设计3组试验,设Modena稀释液+庆大霉素(2×10~5 IU)+复方新诺明(2.5×10~4 IU)为处理1,Modena基础液上分别添加0.5、1、5和10mmol/L的Tau为处理2,处理1配伍处理2则为处理3,每24h检测精子活率、顶体完整率、丙二醛(MDA)浓度以及总抗氧化能力(TAOC),分析Tau与抗生素复合剂对猪精液常温保存效果的影响。结果表明,处理3效果最好,处理2次之,处理1最差,各处理间差异显著,当处理3中Tau的添加量为5mmol/L时,猪精液保存5d后,精子活率为64.11%,顶体完整率为72.10%,T-AOC活性和MDA浓度分别为4.76U/mL和1.42nmol/mL。因此,Tau与抗生素复合剂可以提高猪精液的常温保存效果,最适添加浓度为5mmol/L。     

动物防御素研究进展

防御素(defensin)是一族具有广谱抗微生物和细胞毒活性的多肽,是天然免疫的重要介质,属内源性抗生素肽(endogenous antibiotic peptides)家族.介绍了动物防御素的分布、结构、表达、作用机理等方面的内容.     

Actinomycin Binding to DNA: Inability of a DNA Containing Guanine To Bind Actinomycin D

Polydeoxy (adenylyl-thymidylyl-cytidylyl)* polydeoxy(guanylyl-adenylyl-thymidylyl), a double-stranded DNA polmer of high molecular weight containing 33 percent guanine plus cytosine, binds little or no actinomycin D, as measured by five different techniques. In contrast, the sequence isomer of this DNA, polydeoxy (thymidylyl-adenylyl-cytidylyl)-polydeoxy(guanylyl-thymidylyl-adenylyl), does bind the antibiotic. Thus, the presence of guanine in a DNA is not a sufficient requisite for the binding of actinomycin D.     

Antibiotic-induced inhibition of pheromone synthesis in a bark beetle

Ingestion of diet containing streptomycin inhibited the conversion of myrcene, a host plant terpene, to the male-specific pheromones ipsenol and ipsdienol in 1ps paraconfusus. Synthesis of cis-verbenol, which is not a sex-specific pheromone, from the host plant terpene (-)-alpha-pinene and other metabolites from these two terpenes was not inhibited by the antibiotic.     

开花调控基因BpMADS4无抗性表达载体的构建

为了构建含有报告基因GFP基因和BpMADS4基因的无抗性双元表达载体pCAMBIA1302-GFP-Bp,参考已发表的欧洲白桦（Betula pendula）开花调控基因（BpMADS4）序列,利用逆转录-聚合酶链式反应（RT-PCR）从欧洲白桦的幼嫩花序中克隆得到促进开花的BpMADS4基因。将该基因替换pCAMBIA1302载体中的潮霉素抗性基因,构建含有报告基因GFP基因和BpMADS4基因的无抗性双元表达载体pCAM-BIA1302-GFP-Bp。结果表明：成功构建了无抗性双元表达载体pCAMBIA1302-GFP-Bp。该表达载体在苹果遗传转化中不使用抗生素筛选,可以解决抗生素抗性筛选降低苹果转基因转化效率的问题以及转基因苹果中选择标记基因造成的生物安全性问题,用该载体转化苹果可以缩短苹果童期,有效提高其育种效率。本研究为筛选对环境安全的、具有易成花特性的苹果新种质奠定了基础。     

表达两种不同外膜蛋白的禽大肠杆菌融合菌株的构建

对从陕西省主要养鸡地区病死鸡及鸡胚中分离的致病性禽源大肠杆菌,用SDS-PAGE进行Omp分型。采用不同浓度溶菌酶作用不同时间裂解细胞壁,制备原生质体,在高渗平板上再生培养;在药敏试验的基础上,结合Omp型分析,选择具有交叉耐药且Omp型不同的菌株,利用PEG诱导进行原生质体融合,在含双抗的高渗平板上筛选融合株,对不同代次的融合株分别进行革兰氏染色镜检、生化鉴定、血清型鉴定和Omp型分析。结果表明,所构建的融合菌株染色镜检和生化特性均符合大肠杆菌特征,75%融合子可凝集2亲本的O抗原,25%凝集其中1个亲本的O抗原;外膜蛋白型表现为1个或2个亲本的带型,但都有不同程度的增强。经稳定性检验证明其为遗传学上稳定的融合子。     

中国和欧美国家养殖业抗生素使用负面问题及管理政策比较

畜禽养殖中抗生素的使用无疑推动了该产业的快速发展,但也引起了其耐药性威胁公共健康和养殖业可持续发展的担忧。本文系统梳理了兽用抗生素的使用,抗生素负面问题的显现、认知和管理政策的变化,分析了中国、美国、欧盟管理政策的差异及其背后的原因。本文认为,我国在推进农业现代化进程中：应长期关注养殖业中抗生素使用可能带来的负面问题,加快生产方式的绿色转型;应提升公众对抗生素耐药性的认识,引导公众参与遏制耐药性发展的行动;应考虑我国的发展目标,吸取美国因利益集团阻挠而反应缓慢的教训,综合考虑成本和收益,发挥我国的体制优势,找到政策上的最优解;支持、推动全球抗生素管理的国际协同行动。     

荧光光谱技术在肉类中抗生素残留检测中的应用研究进展

肉类中抗生素残留超标容易对人体健康造成一些潜在危害,其已成为影响肉类食用安全品质的一个重要因素。荧光光谱技术作为一种快速检测技术在肉类中抗生素残留检测当中受到了越来越多的重视。介绍了荧光及荧光发光原理,综述了抗生素检测中的主要荧光发光方法,以及同步荧光光谱、三维荧光光谱、表面增强荧光等常用的荧光光谱技术及其在肉类中抗生素残留检测中的应用情况,并对其应用前景进行了展望。     

农用抗生素对烟草赤星病防治效果的研究

通过离体烟叶来皿叠培养筛选模型得到的链霉菌Ｓ－１０菌株所产生的农用抗生素对烟草赤星病菌具有很强的抑制作用。用牛津小杯法测定的抑菌圈直径可达３．２０ｃｍ。对抗生素进行了抗菌谱,ＰＨ纸色谱,捷允八溶剂系统纸色谱等试验,初步鉴定该抗生素为碱性水溶性广谱抗生素。     

基因枪介导转hpa99基因小麦的获得和检测

为了探究来源于水稻细条斑病菌(Xanthomonas oryzae pv.oryzicola)的hpa99基因改良小麦抗赤霉病的可行性,以小麦幼胚诱导的愈伤组织为转化受体,以磷酸甘露糖异构酶(phosphomannose isomerase,PMI)作为选择标记,通过基因枪法将带有hpa99基因的表达质粒AF234296导入普通小麦品种山农15。2～3次PMI筛选后,获得38株再生植株。通过氯酚红(chlorophenol red,CPR)染色以及PMI基因和目的基因hpa99的PCR检测,获得7株阳性植株,转化率为0.108%。结果表明,hpa99已整合到小麦基因组中,获得了无抗生素标记基因的转基因小麦。     

农杆菌介导的栓皮栎体胚遗传转化初步研究

为了建立栓皮栎体胚遗传转化体系,以未成熟合子胚诱导的体胚为转化受体,采用带有pCAMBIA1301质粒的根癌农杆菌菌株EHA105,其中含有hptⅡ和GUS基因,研究不同因子对遗传转化的影响。结果表明：选择培养中适宜的抗生素浓度为300 mg/L头孢霉素和30 mg/L潮霉素。预培养时间为15 d时,转化率达91.7%;菌液浓度为OD₆₀₀=0.5,乙酰丁香酮浓度为200 μmol/L时,转化率可达100%。在添加潮霉素的选择培养基中培养4周后,组织化学染色和PCR检测证实,GUS报告基因已经转入到体胚中并正常表达。     

井岗霉素在天蓝色链霉菌M145中的异源表达

井岗霉素对水稻纹枯病有良好的防治作用,在我国得到广泛使用。通过结合转移的方法把包含井岗霉素生物合成全基因簇的柯斯质粒9A2导入天蓝色链霉菌M145(Streotpmyces coelicolor M145)中,然后将含有导入质粒的菌株M145/9A2进行发酵,对发酵液进行生物测定的结果显示此菌株发酵液对指示菌水稻纹枯菌有抑制作用,进一步质谱检测结果表明,在天蓝色链霉菌中异源表达井岗霉素成功。     

Construction of a Fosmid genomic library of Streptomyces roseoflavus Men-myco-93-63

To clone the antibiotic biosynthesis gene cluster of Streptomyces roseoflavus Men-myco-93-63, we constructed a Fosmid genomic library. The genomic DNA of the strain Men-myco-93-63 was isolated by the modified CTAB procedure, and the size of most genomic DNA fragments was larger than 150 kb. Then, a Fosmid genomic library containing more than 6000 clones was constructed. The average size of the inserted DNA in recombinant plasmids was 38.1 kb, and the probability of harboring any gene in the genome of the strain Men-myco-93-63 was 99.99%. The library coverage was at least a 10-fold genome equivalent. Therefore, the constructed Fosmid library meets the requirements as a standard genomic library     

链霉菌S024所产生的抗菌活性物质的研究

[目的]为更好满足放线菌对农业生产发展的要求。[方法]从土壤中分离到一批放线菌,从中筛选出1株高活性菌株S024,对其产生的抗生素进行初步研究。[结果]该菌株产生的抗生素抑菌谱广,尤其对小麦纹枯病,小麦根腐病等真菌病害有很好的防治效果。对酸碱稳定,在pH值3.0~9.0中间稳定。温度耐受性较强,在适宜pH条件下,100℃处理0.5 h,其活性基本不变。[结论]对抗生素性质初步分析表明该抗生素为一种水溶性、强极性抗生素,根据性质推断此抗生素属于氨基糖苷类的抗生素。     

土霉素诱导土壤可培养细菌产生耐药性的研究

通过在土壤中反复添加不同浓度的土霉素进行预处理,采用平板稀释法测定了土壤细菌在含土霉素培养基及普通培养基上的生长情况,用耐药菌发生率作为指标评价了土霉素污染浓度和污染次数对土壤抗土霉素可培养细菌的影响。结果表明,土霉素污染可诱导土壤土霉素抗性细菌的产生,耐药菌发生率与土霉素添加浓度和添加次数有关。当土霉素添加浓度大于4 mg/kg时,一次处理就可诱导土壤可培养细菌产生耐药性;当土霉素浓度为3 mg/kg时,二次处理可诱导土壤可培养细菌产生耐药性;当土霉素浓度为2 mg/kg时,需要5次处理才可诱导土壤可培养细菌产生耐药性;而当土霉素浓度为1 mg/kg时,5次处理均没有明显诱导土壤可培养细菌产生耐药性。研究结果表明,长期施用含土霉素的畜禽粪可导致土壤土霉素抗性细菌的形成和发展。     

A convergent enantioselective route to structurally diverse 6-deoxytetracycline antibiotics

Complex antibiotics based on natural products are almost invariably prepared by semisynthesis, or chemical transformation of the isolated natural products. This approach greatly limits the range of accessible structures that might be studied as new antibiotic candidates. Here we report a short and enantioselective synthetic route to a diverse range of 6-deoxytetracycline antibiotics. The common feature of this class is a scaffold of four linearly fused rings, labeled A through D. We targeted not a single compound but a group of structures with the D ring as a site of structural variability. A late-stage, diastereoselective C-ring construction was used to couple structurally varied D-ring precursors with an AB precursor containing much of the essential functionality for binding to the bacterial ribosome. Five derivatives were synthesized from benzoic acid in yields ranging from 5 to 7% over 14 to 15 steps, and a sixth, (-)-doxycycline, was synthesized in 8.3% yield over 18 steps.