22 resultados para S. AUREUS

em Chinese Academy of Sciences Institutional Repositories Grid Portal


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A 115 days feeding trial was conducted to evaluate the effect of dietary cyanobacteria on growth, microcystins (MCs) accumulation in hybrid tilapia (Oreochromis niloticus x O. aureus) and the recovery when the fish were free of cyanobacteria. Three experimental diets were formulated: the control (cyanobacteria free diet); one test diet with cyanobacteria from Lake Taihu (AMt 80.0 mu g MCs g(-1) diet) and one with cyanobacteria from Lake Dianchi (AMd, 410.0 rho g MCs g(-1) diet). Each diet was fed to fish for 60 days and then all fish were free of cyanobacteria for another 55 days. A significant increase in feeding rate (FR) was observed in fish fed AMd diet after a first 30-day exposure (1(st) EP), and in fish fed both AMt diet and AMd diet after a second 30-day exposure (2(nd) EP). Specific growth rates (SGR) of fish fed AMt diet and AMd diet were both obviously affected after the first 30-day exposure, but SGR was only significantly affected in fish fed AMt diet after the second 30-day exposure. After a 55-day recovery, there were no significant differences among diets in the indices mentioned above. Much higher concentrations of MCs were accumulated in tissues of all fish exposed to cyanobacteria. After the 55-day recovery, MC concentrations in fish tissues were significantly lower than those on day 60. (C) 2009 Elsevier Ltd. All rights reserved.

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Bacterial lipoproteins (LP) are a family of cell wall components found in a wide variety of bacteria. In this study, we characterized the response of HUCL, a telomerase-immortalized human corneal epithelial cell (HCEC) line, to LP isolated from Staphylococcus (S) aureus. S. aureus LP (saLP) prepared by Triton X-114 extraction stimulated the activation of NF-kappa B, JNK, and P38 signaling pathways in HUCL cells. The extracts failed to stimulate NF-kappa B activation in HUCL cells after lipoprotein lipase treatment and in cell lines expressing TLR4 or TLR9, but not TLR2, indicating lipoprotein nature of the extracts. saLP induced the up-regulation of a variety of inflammatory cytokines and chemokines (IL-6, IL-8, ICAM-1). antimicrobial molecules (hBD-2, LL-37, and iNOS), and homeostasis genes (Mn-SOD) at both the mRNA level and protein level. Similar inflammatory response to saLP was also observed in primarily cultured HCECs using the production of IL-6 as readout. Moreover, TLR2 neutralizing antibody blocked the saLP-induced secretion of IL-6, IL-8 and hBD2 in HUCL cells. Our findings suggest that saLP activates TLR2 and triggers innate immune response in the cornea to S. aureus infection via production of proinflammatory cytokines and defense molecules. (C) 2007 Elsevier Ltd. All rights reserved.

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Two 8-week growth trials were conducted to determine the effect of continuous (CF) versus 2 meals day(-1) (MF) feeding and 30% starch versus 30% glucose diets on the carbohydrate utilization of 9.0-g white sturgeon and 0.56-g hybrid tilapia. The two trials were conducted under similar conditions except that sturgeon were kept at 18.5 degrees C in a flow-through system and tilapia were kept at 26 degrees C in a recirculating system. Significantly (P less than or equal to 0.05) higher specific growth rate (SGR), feed efficiency (FE), protein efficiency ratio (PER), body lipid content and liver glucose-6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconate dehydrogenase (6PGDH) activities were observed in the CF than MF sturgeon. Only SGR, FE and PER were higher in sturgeon fed the starch than the glucose diets. Only higher liver G6PDH and malic enzyme (ME) activities were observed in the CF than MF tilapia but higher SGR, FE, PER and liver G6PDH, 6PGDH and ME activities were observed in tilapia fed the starch diet than those fed the glucose diet. This suggested that carbohydrate utilization by sturgeon was more affected by feeding strategy whereas tilapia was more affected by carbohydrate source. Furthermore, white sturgeon can utilize carbohydrates better than hybrid tilapia regardless of feeding strategy and carbohydrate source.

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Amphibian skin is a rich resource of antimicrobial peptides like maximins and maximins H from toad Bombina maxima. A novel cDNA clone encoding a precursor protein that comprises maximin 3 and a novel peptide. named maximin H5. was isolated from a skin cDNA library of B. maxima. The predicted primary structure of maximin H5 is ILGPVLGLVSDTLDDVLGIL-NH2,. Containing three aspartate residues and no basic amino acid residues. maximin H5 is characterized by an anionic property. Different from cationic maximin H peptides. only Gram-positive strain Staphylococcus aureus was sensitive to maximin H5. while the other bacteria] and fungal strains tested ere resistant to it. The presence of metal ions. like Zn2+ and Mg2+, did not increase its antimicrobial potency. Maximin H5 represents the first example of potential anionic antimicrobial peptides from amphibians, The results provide the first evidence that. together kith cationic antimicrobial peptides. anionic antimicrobial peptides may also exist naturally as part of the innate defense system. (C), 2002 Elsevier Science (USA). All rights reserved.

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While conducting experiments to investigate antimicrobial peptides of amphibians living in the Yunnan-Sichuan region of southwest China, a new family of antimicrobial peptides was identified from skin secretions of the rufous-spotted torrent frog, Amolops loloensis. Members of the new peptide family named amolopins are composed of 18 amino acids with a unique sequence, for example, NILSSIVNGINRALSFFG. By BLAST search, amolopins did no show similarity to any known peptides. Among the tested microorganisms, native and synthetic peptides only showed antimicrobial activities against Staphylococcus aureus ATCC2592 and Bacillus pumilus, no effects on other microorganisms. The CD spectroscopy showed that it adopted a structure of random combined with beta-sheet in water, Tris-HCl or Tris-HCl-SDS. Several cDNAs encoding amolopins were cloned from the skin cDNA library of A. loloensis. The precursors of amolopin are composed of 62 amino acid residues including predicted signal peptides, acidic propieces, and mature antimicrobial peptides. The preproregion of amolopin precursor comprises a hydrophobic signal peptide of 22 residues followed by an 18 residue acidic propiece which terminates by a typical prohormone processing signal Lys-Arg. The preproregions of precursors are very similar to other amphibian antimicrobial peptide precursors but the mature amolopins are different from other antimicrobial peptide families. The remarkable similarity of preproregions of precursors that give rise to very different antimicrobial peptides in distantly related frog species suggests that the corresponding genes form a multigene family originating from a common ancestor. (C) 2008 Elsevier Masson SAS. All rights reserved.

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In the present study, EA-CATH1 and EA-CATH2 were identified from a constructed lung cDNA library of donkey (Equus asinus) as members of cathelicidin-derived antimicrobial peptides, using a nested PCR-based cloning strategy. Composed of 25 and 26 residues, respectively, EA-CATH1 and EA-CATH2 are smaller than most other cathelicidins and have no sequence homology to other cathelicidins identified to date. Chemically synthesized EA-CATH1 exerted potent antimicrobial activity against most of the 32 strains of bacteria and fungi tested, especially the clinically isolated drug-resistant strains, and minimal inhibitory concentration values against Gram-positive bacteria were mostly in the range of 0.3-2.4 mu g center dot mL-1. EA-CATH1 showed an extraordinary serum stability and no haemolytic activity against human erythrocytes in a dose up to 20 mu g center dot mL-1. CD spectra showed that EA-CATH1 mainly adopts an alpha-helical conformation in a 50% trifluoroethanol/water solution, but a random coil in aqueous solution. Scanning electron microscope observations of Staphylococcus aureus (ATCC2592) treated with EA-CATH1 demonstrated that EA-CATH could cause rapid disruption of the bacterial membrane, and in turn lead to cell lysis. This might explain the much faster killing kinetics of EA-CATH1 than conventional antibiotics revealed by killing kinetics data. In the presence of CaCl2, EA-CATH1 exerted haemagglutination activity, which might potentiate an inhibition against the bacterial polyprotein interaction with the host erythrocyte surface, thereby possibly restricting bacterial colonization and spread.

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A chymotrypsin inhibitor, designated NA-CI, was isolated from the venom of the Chinese cobra Naja atra by three-step chromatography. It inhibited bovine (x-chymotrypsin with a K-i of 25 nM. The molecular mass of NA-CI was determined to be 6403.8 Da by matrix-assisted laser-desorption ionization time-of-flight (MALDI-TOF) analysis. The complete amino acid sequence was determined after digestion of S-carboxymethylated inhibitor with Staphylococcus aureus V8 protease and porcine trypsin. NA-CI was a single polypeptide chain composed of 57 amino acid residues. The main contact site with the protease (PI) has a Phe, showing the specificity of the inhibitor. NA-CI shared great similarity with the chymotrypsin inhibitor from Naja naja venom (identities = 89.5%) and other snake venom protease inhibitors. (C) 2003 Elsevier Inc. All rights reserved.

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蛇毒和蜂毒是提供药理学活性分子的丰富来源,它们富含肽和蛋白,包括一 些酶类和毒素。 丝氨酸蛋白酶抑制剂广泛存在于动物、植物和微生物体内,参与许多重要的 生理过程,如血液凝集、纤维蛋白溶解、细胞凋亡、发育以及炎症反应和补体活 化等(van Gent D. et al., 2003)。通过凝胶过滤、离子交换和反向高压液相色谱, 我们从金环蛇毒液中纯化得到一种天然的丝氨酸蛋白酶抑制剂,命名为 bungaruskunin。并且从该蛇的毒腺cDNA 文库中克隆到了它的核苷酸序列。 bungaruskunin 预测的前体由83 个氨基酸组成,包括含有24 个氨基酸的信号肽 和含有59 个氨基酸的成熟肽。它与一种由红腹伊澳蛇(Pseudechis porphyriacus 的cDNA 预测到的丝氨酸蛋白酶抑制剂blackelin 具有最大相似性,达64%。 Bungaruskunin 是一种Kunitz 型的蛋白酶抑制剂,具有一个保守的Kunitz 结构域, 能够抑制胰蛋白酶、胰凝乳蛋白酶和弹性蛋白酶。通过对金环蛇毒腺cDNA 文库 的筛选,我们还得到了另外两条β-bungarotoxin B 链,Bungaruskunin 的整体结 构与β-bungarotoxin B 链相似,特别是它们都具有高度保守的信号肽序列。这些 发现强烈地表明蛇毒Kunitz/BPTI 蛋白酶抑制剂与神经毒性的类似物可能起源于 共同的祖先。 肥大细胞脱粒肽是从膜翅目昆虫的毒液中鉴别出的一个小肽家族,是一种具 有潜在的药物治疗作用的诱导活性分子(Xueqing Xu et al., 2006)。来源于蜂类的 缓激肽样的类似物vespakinin 家族是一种具有调节和激素功能的活性成分,与哺 乳动物和两栖动物的缓激肽类似(Nakajima T., 1984)。本研究对三种胡蜂的 毒液进行了一系列的活性检测,发现黑尾胡蜂的蜂毒对白色念珠菌Candida albicans 和金黄色葡萄球菌 Staphylococcus aureus 有抑制作用。凹纹胡蜂和黑尾 胡蜂的蜂毒具有微弱的磷酯酶A2 活性。通过凝胶过滤和反向高压液相色谱,没 有得到相关的活性组分。通过对三种胡蜂毒腺cDNA 文库的筛选,我们得到了2 条来源于黑尾胡蜂的核苷酸序列,Blast 分析表明,其中一条编码类似肥大细胞 脱粒肽,但未克隆到全长,序列比对结果显示其与来源于大胡蜂(Vespa magnifica) 的Mastoparan-like peptide 12c precursor(GenBank accession A0SPI0)的核苷酸序 列相似性达98%(Xueqing Xu et al., 2006);另一条编码缓激肽类似物,命名为 Hw-bradykinin,序列比对结果显示其与来源于大胡蜂(Vespa magnifica)的 vespakinin-M precursor(GenBank accessionABG75944)的核苷酸相似率达96% (Zouhong Zhou et al., 2006)。

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本文采用几何形态测量的方法对产自和政地区的鬣狗科Hyaenictitherium, Ictitherium, Adcrocuta和旁鬣狗科Dinocrocuta的材料进行研究,分析了这4个属保存完好的头骨标本的侧面形态。与东非大草原现生食肉动物的头骨整体形态分布的比较和分析表明,和政的鼬鬣狗(Ictitherium)和鬣型鼬鬣狗(Hyaenictitherium)的头骨形状分布介于现生斑鬣狗(Crocuta crocuta)、犬科猎狗(Lycaon pictus)和金豺(Canis aureus)之间,为二者似豺生态形态的解释提供了几何形态测量证据。再者,上述两属化石鬣狗的形状分布与现生斑鬣狗的幼年个体形状重叠,表明现生斑鬣狗头骨的发育机制可能是在鼬鬣狗祖先类型的异速生长规律基础上的持续发育,进而演化出现有的粗壮形态。此外,巨鬣狗(Dinocrocuta)和副鬣狗(Adcrocuta)的头骨形状与现生的斑鬣狗在几何形态测量空间内有普遍重叠的现象,指示了这些异时出现的种类具有相似的生态形态,因而可能占据相近的生态位。结果还显示巨鬣狗和斑鬣狗的幼年个体形状相近,以及两者从幼年到成年发育的形状变化过程也具有相似的规律。因而,巨鬣狗和斑鬣狗之间的趋同演化不仅表现在成年头骨的粗壮程度上,而且在幼年发育模式中也存在平行演化现象。现生发育学与行为生态学已经证实,相对其他大型食肉动物,现生斑鬣狗发育粗壮头骨形态的机制不是以增速生长,而是以延长发育期来实现的。由此推断,巨鬣狗的发育期有可能和现生鬣狗相当(35个月),也可能由于具有相对粗壮和巨大的头骨形态,其发育期会延长些。当然,这个新解释仍需要更多的化石数据和发育研究来证实。

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链霉菌是十分重要的一类放线菌,绝大多数的抗生素都由该类细菌产生。毛壳属真菌是一类重要的丝状真菌,从中也发现有很多结构新颖、活性独特的活性物质。因此本论文对两株链霉菌的活性成分及一株金毛壳菌的次生代谢产物进行了研究。 1.从吸水链霉菌(Streptomyces hygroscopicus 1.358)液态发酵产物(乙酸乙酯提取物)中分离得到3个化合物,通过波谱方法鉴定为RK955A (1)、Nigericin(2)、Elaiophylin(3)。以青霉素耐药-金黄色葡萄球菌作为指示菌的抗菌活性测定表明,三者均具有较强抗菌活性。 2.通过抗肿瘤体外活性筛选模型筛选得到得到一株链霉属土壤放线菌,从中分离得到六个化合物:苯乙酰胺(4)、苯丙酰胺(5)、肉桂酰胺(6)、3-(N-(甲酰胺基)乙酰基)吲哚(7)、鸟苷磷酸(8)、鸟苷(9)。 3.从金色毛壳菌(Chaetomium aureus)的固态培养物中分离得到13个化合物,利用波谱方法将其鉴定为:金毛壳菌素A(10)、金毛壳菌素B(11)、Eugenetin(12)、Eugenitol(13)、Chaetoquadrin A(14)、Chaetoquadrin B(15)、Chaetoquadrin G(16)、Chaetoquadrin H(17)、Chaetochromin A(18)、Sterigmatocystin(19)、O-methylsterigmatocystin(20)、3β-羟基-麦角甾-5,7,22-三烯(21)和过氧麦角甾醇(22)。 4.综述了聚醚类抗生素的结构、生物合成、生物活性及作用机理。 The genus Streptomyces (Actinomycetes) is an important group of microbe. Most antibiotics known nowdays are discovered from species of Streptomyces. The fungi of the genus Chaetomium have attracted much attention because various kinds of secondary metabolites with diverse bioactivities have been found from them. Thus, the bioactive compounds from two strains of Streptomyces and the secondary metabolites of Chaetomium aureus were investigated. 1. Three compounds were isolated from the ethyl acetate extract of the fermentation broth of Streptomyces hygroscopicus. They are identified to be elaiophylin (1), nigericin (2), and antibiotic RK955A (3) on the basis of their spectroscopic data. Compounds 1-3 possess antibacterial activities against Staphyloccocus aureus. 2. It was found that the extract of the fermented broth of a strain of Actinomycetes could inhibit some tumor cel lines. Separation of the bioactive fraction led to the isolation of six compounds. They were characterized to be phenylacetamide (4), phenylpropylamide (5), trans-cinnamamide (6), 3- (N- (formylmethyl) acetamide) indole (7), guanylicacid (8), and guanosine (9). 3. From the fermented broth of Chaetomium aureus, 13 compounds were isolated for the first time. They were determined to be chaetomiumycin A (10), chaetomiumycin B (11), eugenetin (12), eugenitol (13), chaetoquadrin A (14), chaetoquadrin B (15), chaetoquadrin G (16), chaetoquadrin H (17), chaetochromin A (18), sterigmatocystin (19), O-methylsterigmatocystin (20), 3β-hydroxyergosta-5, 7, 22-triene (21) and peroxy-ergosterol (22). Compounds 10 and 11 are new ones. 4. Structure, biosynthesis, biological activity, and mechanisms of polyether antibiotics were reviewed.

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本论文以从四川峨嵋山森林土壤中分离筛选获得的一株产抗耐药性活性化合物的链霉菌S227为材料,对发酵液中活性物质的分离纯化及抗耐药性活性进行了研究。 建立了抗耐药性活性的定性、定量检测方法。建立的管蝶法活性定量检测的标准回归方程为:D=4.8229Ln(C)+3.6326 R=0.9972 ;纸片法活性定量检测的标准回归方程为:D=5.5Ln(C)-12.794 R=0.999。 根据建立的样品活性的检测方法,测定了发酵液的初始活性。实验证明活性物质的温度、pH稳定性好。 通过活性的定性、定量追踪方法,分别利用等体积的石油醚、乙酸乙酯、正丁醇在不同的pH梯度下萃取,确定了pH3条件下正丁醇能最大程度的萃取活性物质,说明活性物质极性很大。对正丁醇萃取相经过两次硅胶柱层析及薄层层析分离得到具有抗耐药菌活性的纯化样品S227-4。 经过核磁共振氢谱、碳谱数据分析初步确定S227-4为四聚糖,通过糖的水解实验初步确定S227-4由葡萄糖和半乳糖组成。 纸片法活性检测表明S227-4具有抗耐药菌活性。采用MIC测定法对该样品抗耐药活性进行研究。在证明该样品本身不具有抗菌活性的基础上,以临床分离的耐药性金黄色葡萄球菌为指示菌,考察了该样品与抗生素联合使用时对耐药菌抗生素MIC(最小抑菌浓度)值的影响,结果表明在不影响菌体生长的浓度条件下,该样品能明显降低多株耐药菌对多种抗生素的MIC值,不同程度地恢复所测试耐药菌对相应抗生素的敏感性。如S227-4与青霉素钠联用可以使S. aureus 12352的MIC降低8倍,而与红霉素联用可以使S. aureus 12334的MIC降低128倍。 The purification process and the activity of the anti bacterial drug resistance compounds produced by Streptomyces S227 isolated from the forest soil sample of the Mountain E’MEI in Sichuan Province were studied in this thesis. Quantitative and qualitative activity assay methods of the active compounds were established. The regression equation of the tube method was D=4.8229Ln(C)+3.6326, R=0.9972. The regression equation of the paper method was D=5.5Ln(C)-12.794, R=0.999. According to the established activity assay method, the incipient activity of the broth was evaluated. And it was proved that the stability of the active compounds was good. By quantitative and qualitative activity tracing method, petroleum ether, ethyl acetate and butanol were used to extract the active compounds at different pH. The result showed that butanol was the most effective agent for active component recovery at pH3. From the butanol extraction a purified sample, S227-4, was isolated by silica gel column chromatography and thin-layer chromatography . S227-4 was proved to be a tetra- saccharide by 1H-NMR and 13C-NMR. And its monosaccharides include glucose and galactose by hydrolysate analysis. The anti-drug resistant activity of S227-4 was tested in vitro by MICs assay using different drug resistant Staphylococcus aureus strains isolated clinically. The sample itself showed no anti-microbial activity in growth inhibitory experiment, but when it was used together with different antibiotics, it could remarkably decrease the MICs of different clinically isolated drug-resistant bacterial strains to these antibiotics. For example, when S227-4 was used with penicillin, the MIC of S. aureus 12352 decreased 8 times compared with that when penicillin was used alone. Meanwhile when it was used with erythromycin the MIC of S. aureus 12334 deceased 128 times compared with erythromycin alone.

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To study the effects of radiation sterilization of the electron beam,the three species of microorganisms,Escherichia.coli,Staphylococcus aureus and Proteus vulgaris were irradiated with the electron beam,delivered by the electron accelerator independently developed by the Institute of Modern Physics,Chinese Academy of Sciences,and the changes of superoxide dismutase(SOD) activity of these irradiated microorganisms were also tested.The results indicated that the Staphylococcus aureus were fully radio-sterili...中文摘要:在中国科学院近代物理研究所自行研制的大功率电子加速器上,研究了不同辐照剂量的电子束对大肠杆菌、金黄色葡萄球菌和变形杆菌3种微生物的杀灭效果,同时检测了辐照后菌体超氧化物歧化酶(SOD)活性的变化。结果显示:辐照剂量达到2.0 kGy时,可完全杀灭金黄色葡萄球菌,2.2 kGy时可完全杀灭大肠杆菌和变形杆菌;辐照对3种微生物的SOD活性有较显著的影响。

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The bioactivity screening of fractions from two inter-tidal sponges collected from the north of China Yellow Sea and one sponge collected from the South Chinese Sea was reported in this study. In sponge Hymeniacidon perleve there were 9 fractions out of 15 from CHCl3 extract with anti Staphylococcus aureus activity, 9 fractions out of 19 from BuOH extract with anti Escherichia coli activity, and three fractions from CHCl3 extract which had moderate to strong activity in inhibiting Bacillus subtilis, Candida albicans, and Aspergilus niger. The fractions of Reniochalina sp. showed bioactivity against bacteria and fungi. The fractions of Acanthella acuta Schmidt showed bioactivity against S. aureus and fungi. One compound from H. perleve obtained by the bioactively directing isolation was tested for bioactivity against the human hepatoma cell line Qgy7701 (IC50 10.1 mug/ml), Burkitt's lymphoma cell line Raji (IC50 9.76 mug/ml) and chronic myelogenous leukemia K562 (IC50 1.90 mug/ml). (C) 2003 Elsevier B.V. All rights reserved.

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The poly(vinyl alcohol)/ poly(N-vinyl pyrrolidone) (PVA-PVP) hydrogels containing silver nanoparticles were prepared by repeated freezing-thawing treatment. The silver content in the solid composition was in the range of 0.1-1.0 wt %, the silver particle size was from 20 to 100 nm, and the weight ratio of PVA to PVP was 70 : 30. The influence of silver nanoparticles on the properties of PVA-PVP matrix was investigated by differential scanning calorimeter, infrared spectroscopy and UV-vis spectroscopy, using PVA-PVP films containing silver particles as a model. The morphology of freeze-dried PVA-PVP hydrogel matrix and dispersion of the silver nanoparticles in the matrix was examined by scanning electron microscopy. It was found that a three-dimensional structure was formed during the process of freezing-thawing treatment and no serious aggregation of the silver nanoparticles occurred. Water absorption properties, release of silver ions from the hydrogels and the antibacterial effects of the hydrogels against Escherichia coli and Staphylococcus aureus were examined too. It was proved that the nanosilver-containing hydrogels had an excellent antibacterial ability.

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Biodegradable poly(L-lactide) (PLA) ultrafine fibers containing nanosilver particles were prepared via electrospinning. Morphology of the Ag/PLA fibers and distribution of the silver nanoparticles were characterized. The release of silver ions from the Ag/PLA fibers and their antibacterial activities were investigated. These fibers showed antibacterial activities (microorganism reduction) of 98.5% and 94.2% against Staphylococcus aureus and Escherichia coli, respectively, because of the presence of the silver nanoparticles.