Aminoglycoside antibiotics induce bacterial biofilm formation

Biofilms are adherent aggregates of bacterial cells that form on biotic and abiotic surfaces, including human tissues. Biofilms resist antibiotic treatment and contribute to bacterial persistence in chronic infections. Hence, the elucidation of the mechanisms by which biofilms are formed may assist in the treatment of chronic infections, such as Pseudomonas aeruginosa in the airways of patients with cystic fibrosis. Here we show that subinhibitory concentrations of aminoglycoside antibiotics induce biofilm formation in P. aeruginosa and Escherichia coli. In P. aeruginosa, a gene, which we designated aminoglycoside response regulator (arr), was essential for this induction and contributed to biofilm-specific aminoglycoside resistance. The arr gene is predicted to encode an inner-membrane phosphodiesterase whose substrate is cyclic di-guanosine monophosphate (c-di-GMP)-a bacterial second messenger that regulates cell surface adhesiveness. We found that membranes from arr mutants had diminished c-di-GMP phosphodiesterase activity, and P. aeruginosa cells with a mutation changing a predicted catalytic residue of Arr were defective in their biofilm response to tobramycin. Furthermore, tobramycin-inducible biofilm formation was inhibited by exogenous GTP, which is known to inhibit c-di-GMP phosphodiesterase activity. Our results demonstrate that biofilm formation can be a specific, defensive reaction to the presence of antibiotics, and indicate that the molecular basis of this response includes alterations in the level of c-di-GMP.     

Infection regulates pro-resolving mediators that lower antibiotic requirements

Underlying mechanisms for how bacterial infections contribute to active resolution of acute inflammation are unknown. Here, we performed exudate leukocyte trafficking and mediator-metabololipidomics of murine peritoneal Escherichia coli infections with temporal identification of pro-inflammatory (prostaglandins and leukotrienes) and specialized pro-resolving mediators (SPMs). In self-resolving E. coli exudates (10(5) colony forming units, c.f.u.), the dominant SPMs identified were resolvin (Rv) D5 and protectin D1 (PD1), which at 12?h were at significantly greater levels than in exudates from higher titre E. coli (10(7) c.f.u.)-challenged mice. Germ-free mice had endogenous RvD1 and PD1 levels higher than in conventional mice. RvD1 and RvD5 (nanograms per mouse) each reduced bacterial titres in blood and exudates, E. coli-induced hypothermia and increased survival, demonstrating the first actions of RvD5. With human polymorphonuclear neutrophils and macrophages, RvD1, RvD5 and PD1 each directly enhanced phagocytosis of E. coli, and RvD5 counter-regulated a panel of pro-inflammatory genes, including NF-κB and TNF-α. RvD5 activated the RvD1 receptor, GPR32, to enhance phagocytosis. With self-limited E. coli infections, RvD1 and the antibiotic ciprofloxacin accelerated resolution, each shortening resolution intervals (R(i)). Host-directed RvD1 actions enhanced ciprofloxacin's therapeutic actions. In 10(7) c.f.u. E. coli infections, SPMs (RvD1, RvD5, PD1) together with ciprofloxacin also heightened host antimicrobial responses. In skin infections, SPMs enhanced vancomycin clearance of Staphylococcus aureus. These results demonstrate that specific SPMs are temporally and differentially regulated during infections and that they are anti-phlogistic, enhance containment and lower antibiotic requirements for bacterial clearance.     

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.     

Antibiotics in early life alter the murine colonic microbiome and adiposity

Antibiotics administered in low doses have been widely used as growth promoters in the agricultural industry since the 1950s, yet the mechanisms for this effect are unclear. Because antimicrobial agents of different classes and varying activity are effective across several vertebrate species, we proposed that such subtherapeutic administration alters the population structure of the gut microbiome as well as its metabolic capabilities. We generated a model of adiposity by giving subtherapeutic antibiotic therapy to young mice and evaluated changes in the composition and capabilities of the gut microbiome. Administration of subtherapeutic antibiotic therapy increased adiposity in young mice and increased hormone levels related to metabolism. We observed substantial taxonomic changes in the microbiome, changes in copies of key genes involved in the metabolism of carbohydrates to short-chain fatty acids, increases in colonic short-chain fatty acid levels, and alterations in the regulation of hepatic metabolism of lipids and cholesterol. In this model, we demonstrate the alteration of early-life murine metabolic homeostasis through antibiotic manipulation.     

抗菌含银HA膜的研制

在自然界和很多水生环境中,细菌可以附着在一些不活动的表面并形成菌落和包围菌落的生物膜.反过来这层膜的形成又更容易吸附细菌,尤其是在医用植入设备中,这层膜对人体有毒害作用,并且由于这层膜的保护,抗生素和人体的防御系统都不起作用.含银HA镀膜材料很好地解决了这一问题,并对抗菌医用植入材料引起的感染提供了乐观的前景.对在Ti6AL4V表面镀含银羟基磷灰石(HA)膜形成的抗菌界面加以研究.     

Pseudomonas biofilm formation and antibiotic resistance are linked to phenotypic variation

Colonization of the lungs of cystic fibrosis (CF) patients by the opportunistic bacterial pathogen Pseudomonas aeruginosa is the principal cause of mortality in CF populations. Pseudomonas aeruginosa infections generally persist despite the use of long-term antibiotic therapy. This has been explained by postulating that P. aeruginosa forms an antibiotic-resistant biofilm consisting of bacterial communities embedded in an exopolysaccharide matrix. Alternatively, it has been proposed that resistant P. aeruginosa variants may be selected in the CF respiratory tract by antimicrobial therapy itself. Here we report that both explanations are correct, and are interrelated. We found that antibiotic-resistant phenotypic variants of P. aeruginosa with enhanced ability to form biofilms arise at high frequency both in vitro and in the lungs of CF patients. We also identified a regulatory protein (PvrR) that controls the conversion between antibiotic-resistant and antibiotic-susceptible forms. Compounds that affect PvrR function could have an important role in the treatment of CF infections.     

Global organization of metabolic fluxes in the bacterium Escherichia coli

Cellular metabolism, the integrated interconversion of thousands of metabolic substrates through enzyme-catalysed biochemical reactions, is the most investigated complex intracellular web of molecular interactions. Although the topological organization of individual reactions into metabolic networks is well understood, the principles that govern their global functional use under different growth conditions raise many unanswered questions. By implementing a flux balance analysis of the metabolism of Escherichia coli strain MG1655, here we show that network use is highly uneven. Whereas most metabolic reactions have low fluxes, the overall activity of the metabolism is dominated by several reactions with very high fluxes. E. coli responds to changes in growth conditions by reorganizing the rates of selected fluxes predominantly within this high-flux backbone. This behaviour probably represents a universal feature of metabolic activity in all cells, with potential implications for metabolic engineering.     

强化工业大肠杆菌生长特性的研究

大肠杆菌是制药工业中的重要蛋白质药物生产菌株。然而乙酸积累、中心碳代谢负担过重等不良因素严重制约着大肠杆菌的高密度生长。利用Red同源重组技术,构建icd和ptsG双基因敲除大肠杆菌菌株,以分流碳代谢流,提高碳利用效率,最终增加菌体生物量。同时构建ppc过表达质粒,减少乙酸积累。发酵结果显示,构建的重组菌株产酸量大幅减少,pH值平均升高20%,菌体浓度增加了50%,生长特性得到了强化。     

迷迭香酸的抗菌机理研究

为了了解迷迭香酸的抗菌活性及抗菌机理,采用平板生长抑制法,变性聚丙烯酰胺凝胶电泳法（SDS-PAGE）,聚合酶链式反应（PCR）分析等方法,研究了迷迭香酸对大肠杆菌（Escherichia coli）和金黄色葡萄球菌（Staphylococcus aureus）的抗菌活性及其抗菌机理。结果表明,迷迭香酸对大肠杆菌和金黄色葡萄球菌均有抗菌活性,且对金黄色葡萄球菌的活性大于对大肠杆菌的活性;迷迭香酸可改变细菌细胞膜的通透性,导致还原糖和蛋白质的渗漏而影响细胞代谢,通过抑制DNA聚合酶的活性而影响DNA复制,从而发挥了抑菌作用。     

Energy dependence and reversibility of membrane alterations induced by polyene macrolide antibiotics in Chlorella vulgaris

The requirement of metabolic energy for the interaction of polyene macrolide antibiotics with eukaryotic organisms remains a controversial subject (for review see ref. 1) It has been claimed that the lethal binding of these antibiotics to the sterol target component of the hydrophobic core of the membrane, in accordance with the model of de Kruijff and Demel, is an energy-dependent process. When energy production is reduced by removal of all metabolisable substrates or by adding metabolic inhibitors, polyene binding and antifungal effects are also reduced. Metabolic energy may be required to maintain binding site accessibility or to move antibiotic molecules to the active site. The interaction is also restricted at low temperatures, possibly because of the reduced thermal mobilities of the groups concerned with antibiotic uptake. However, it should be emphasised that the interaction of polyene macrolides with artificial lipid membranes is a purely physicochemical process, although the type of permeability pathways induced are similar to those observed in natural membranes. Using Chlorella vulgaris as a model organism, we demonstrate here that the interaction of polyene macrolides with sensitive cells and the induction of lethal membrane permeability changes are energy-dependent processes or purely physicochemical phenomena, depending on the structure of the antibiotic used.     

两种培养基对蒲公英体外抑菌效果观察

对蒲公英全草水提液与醇提液分别在普通营养培养基和M-H琼脂培养基上进行了大肠杆菌、金黄色葡萄球菌抑茵作用的观察，比较两种培养基对中药体外抑菌效果，结果表明，M-H琼脂培养基对其抑菌作用的检测效果比普通营养培养基灵敏。     

A component of innate immunity prevents bacterial biofilm development

Antimicrobial factors form one arm of the innate immune system, which protects mucosal surfaces from bacterial infection. These factors can rapidly kill bacteria deposited on mucosal surfaces and prevent acute invasive infections. In many chronic infections, however, bacteria live in biofilms, which are distinct, matrix-encased communities specialized for surface persistence. The transition from a free-living, independent existence to a biofilm lifestyle can be devastating, because biofilms notoriously resist killing by host defence mechanisms and antibiotics. We hypothesized that the innate immune system possesses specific activity to protect against biofilm infections. Here we show that lactoferrin, a ubiquitous and abundant constituent of human external secretions, blocks biofilm development by the opportunistic pathogen Pseudomonas aeruginosa. This occurs at lactoferrin concentrations below those that kill or prevent growth. By chelating iron, lactoferrin stimulates twitching, a specialized form of surface motility, causing the bacteria to wander across the surface instead of forming cell clusters and biofilms. These findings reveal a specific anti-biofilm defence mechanism acting at a critical juncture in biofilm development, the time bacteria stop roaming as individuals and aggregate into durable communities.     

P pili in uropathogenic E. coli are composite fibres with distinct fibrillar adhesive tips.

Escherichia coli is a frequent cause of several common bacterial infections in humans and animals, including urinary tract infections, bacteraemia and bacteria-related diarrhoea and is also the main cause of neonatal meningitis. Microbial attachment to surfaces is a key event in colonization and infection and results mainly from a stereochemical fit between microbial adhesins and complementary receptors on host cells. Bacterial adhesins required for extracellular colonization by Gram-negative bacteria are often minor components of heteropolymeric fibres called pili which must be oriented in an accessible manner in these structures to be able to bind to specific receptor architectures. P pili mediate the binding of uropathogenic E. coli to a digalactoside receptor determinant present in the urinary tract epithelium. We report here that the adhesin is a component of distinct fibrillar structures present at the tips of the pili. These virulence-associated tip fibrillae are thin, flexible polymers composed mostly of repeating subunits of PapE that frequently terminate with the alpha-D-galactopyranosyl-(1-4)-beta-D-galactopyranose or Gal alpha (1-4)Gal binding PapG adhesin.     

幽门螺杆菌与消化性溃疡,消化不良症的关系

幽门螺杆菌与消化性溃疡、消化不良症的关系①林炳珍杨善民颜江华张长弓（厦门大学抗癌研究中心厦门３６１００５）幽门螺杆菌（Ｈｅｌｉｃｏｂａｃｔｅｒｐｙｌｏｒｉ，ＨＰ）以很高的人群感染率及与慢性胃炎和消化性溃疡的高度相关性引起国内外学者高度重视［１，２］．...     

黔产金银花提取物抑菌作用的初步研究

研究黔产金银花醇提物和水提物对金黄色葡萄球菌和大肠杆菌的抑菌活性,结果表明,黔产金银花醇提物和水提物对金黄色葡萄球菌和大肠杆菌均有一定的抑菌作用,其中,醇提物对金黄色葡萄球菌、大肠杆菌的最低抑菌浓度分别为250mg/mL、250mg/mL,水提物对金黄色葡萄球菌、大肠杆菌的最低抑菌浓度分别为62.5mg/mL、125mg/mL。     

内蒙古额吉淖尔盐碱湖嗜盐古菌的抗生素抗性及分泌的嗜盐菌素的抑菌性

本实验室从内蒙古额吉淖尔湖盐碱土样分离得到一株属于Natrinema属的嗜盐古菌。本文对其进行了抗生素抗性实验以及对其产生的嗜盐菌素进行了抑菌性研究。结果表明诺氟沙星对该菌具有较强的抑菌作用,头孢氨苄对该菌具有微弱的抑菌作用。通过测定抑菌圈的大小表明该菌株对抗生素具有一定的抗性。该菌株所生产的嗜盐菌素对蛋白酶K 较敏感,对大肠杆菌和金黄色葡萄球菌具有抑菌作用。     

高压脉冲电场处理瓶装饮用水的研究

探讨高压脉冲电场处理瓶装水中大肠杆菌、啤酒酵母、青霉、金黄色葡萄球菌和沙门氏菌的杀菌效果.结果表明,PEF可以有效地杀灭瓶装水中常见微生物,其中大肠杆菌、啤酒酵母和青霉菌经48.39 kV/cm场强的PEF处理,在不同时间之后均未有活菌检出,金黄色葡萄球菌和沙门氏菌在此场强下处理经295μs之后,活菌数分别减少了6.6...     

CHMA医用广谱抗菌耐洗涤织物的应用性能测试及临床应用

制备了CHMA医用广谱抗菌耐洗涤织物。实验表明:该抗菌织物具有抗菌谱广、抗菌作用强、作用时间短和耐洗涤等特点。对金黄色葡萄球菌、大肠杆菌和绿脓杆菌等五十多种细菌具有显著的杀灭和抑制作用。皮肤刺激和过敏试验以及临床应用表明CHMA医用广谱抗菌耐洗涤织物可供人体安全使用。在医疗和公共场所采用该织物制成的各种医用辅料、床单、工作服等,可显著地降低细菌感染率和交叉感染率。     

几种浮游单细胞藻类磷代谢的初步研究

用３２ＰＯ４３－同位素稀释法研究了单种一次培养的几种浮游单细胞藻类的磷代谢．小球藻（Ｃｈｌｏｒｅｌｌａｓｐ．）和一种绿藻对ＰＯ４３－的最大吸收速率（Ｖｍａｘ）与细胞磷含量成负相关，半饱和速率常数（Ｋｓ）与藻类的种类相关．光照与黑暗对中华盒形藻（ＢｉｄｄｕｌｐｈｉａｓｉｎｅｎｓｉｓＧｒｅｖｉｌｅ）和金藻（Ｄｉｃｒａｔｅｒｉａｓｐ）ＰＯ４３－的吸收和再生代谢无明显影响，其昼夜变化是藻类细胞新陈代谢的反映；处于细胞周期Ｇ１期与Ｓ期的藻类有较高的磷吸收和再生代谢速率，而Ｇ２期与Ｍ期的磷代谢速率较低．藻类磷代谢的动力学及其昼夜变化是围绕着对营养盐磷最大程度地利用和维持藻类持续生长的机制展开的．     

Biodiversity improves water quality through niche partitioning

Excessive nutrient loading of water bodies is a leading cause of water pollution worldwide, and controlling nutrient levels in watersheds is a primary objective of most environmental policy. Over the past two decades, much research has shown that ecosystems with more species are more efficient at removing nutrients from soil and water than are ecosystems with fewer species. This has led some to suggest that conservation of biodiversity might be a useful tool for managing nutrient uptake and storage, but this suggestion has been controversial, in part because the specific biological mechanisms by which species diversity influences nutrient uptake have not been identified. Here I use a model system of stream biofilms to show that niche partitioning among species of algae can increase the uptake and storage of nitrate, a nutrient pollutant of global concern. I manipulated the number of species of algae growing in the biofilms of 150 stream mesocosms that had been set up to mimic the variety of flow habitats and disturbance regimes that are typical of natural streams. Nitrogen uptake rates, as measured by using (15)N-labelled nitrate, increased linearly with species richness and were driven by niche differences among species. As different forms of algae came to dominate each unique habitat in a stream, the more diverse communities achieved a higher biomass and greater (15)N uptake. When these niche opportunities were experimentally removed by making all of the habitats in a stream uniform, diversity did not influence nitrogen uptake, and biofilms collapsed to a single dominant species. These results provide direct evidence that communities with more species take greater advantage of the niche opportunities in an environment, and this allows diverse systems to capture a greater proportion of biologically available resources such as nitrogen. One implication is that biodiversity may help to buffer natural ecosystems against the ecological impacts of nutrient pollution.