大花黄牡丹内生菌的分离鉴定及其抗菌活性菌株的筛选

采用研磨法从健康大花黄牡丹的根、茎、叶柄、叶和种子中进行菌种分离,依据其形态、培养特征及其他生物学特性对菌株进行初步鉴定;采用平板对峙法对分离的内生菌进行拮抗试验研究,并对强活性菌株进行16S rD-NA序列鉴定,以明确大花黄牡丹内生菌的种类,筛选对农作物病害有抑制作用的菌株.结果表明:(1)获得内生真菌188株,鉴定为10个属,以短蠕孢属(50％)、青霉孢属(18.6％)和曲霉孢属(12.3％)为优势种群.获得内生放线菌145株,以链霉菌属(98.6％)为优势种群.表明大花黄牡丹内生菌在数量和种类上存在极丰富的多样性,同时在不同组织存在一定的差异性.(2)抑菌试验结果显示,21.6％真菌对指示菌有抑菌作用,抑菌圈直径最大为10mm;27.8％放线菌对指示菌有抑菌作用,其中菌株PND31的抑菌活性较强,抑菌谱较广.(3)16S rDNA序列鉴定显示,菌株PND31与链霉菌属聚在一起,初步归为链霉菌属一个种.     

无患子内生真菌抗菌筛选与鉴定

目的筛选和鉴定具有抗菌活性的无患子内生真菌,为进一步探索其抗菌成分提供菌源。方法采用拮抗试验和体外抗菌测试方法,以大肠埃希菌(Escherichia coli)、金黄色葡萄球菌(Staphylococcus aureus)和白假丝酵母菌(Candida albicans)为指示菌,首次研究6株无患子内生真菌(HY11-1、HY11-2、P11、O、ye4、ye5)的抗菌特点。通过形态学观察和18SrDNA序列分析,对活性菌株进行菌种鉴定。结果菌株HY11-1、ye5和P11具有较强的抗菌能力,其中菌株HY11-1对金黄色葡萄球菌抑菌效果最显著,确定菌株HY11-1、ye5、P11分别为Alternaria alternate、Aspergillus ochraceus和Penicillium chrysogenum。结论 3株无患子内生真菌A.alternate HY11-1、A.ochraceus ye5和P.chrysogenum P11具有较强的抗菌效果,具有进一步研究的价值。     

海南粗榧内生真菌抗肿瘤抗菌活性的筛选

对72株海南粗榧(Cephalotaxus hainanensis Li)内生真菌进行了抗肿瘤和抗菌活性筛选。结果显示, 有9株内生真菌至少对一种指示瘤株具有细胞毒活性, 5株内生真菌对金黄色葡萄球菌有较强的抑菌活性, 1株内生真菌对辣椒疫霉有抑制作用。这表明海南粗榧内生真菌是寻找有价值的生物活性成分的潜在资源, 其生物活性成分值得进一步研究。     

Inhibitory Effect of Paenibacillus polymyxa GBR-462 on Phytophthora capsici Causing Phytophthora Blight in Chili Pepper

In the present work 25 strains of Paenibacillus polymyxa isolated from rotted ginseng roots were screened for their antimicrobial activity against Phytophthora capsici in vitro . Based on antimicrobial activity, 15 strains categorized as strongly antimicrobial, among them GBR-462 was found as the most active, and five strains each as weekly antimicrobial and no antimicrobial. Antimicrobial activity was influenced by the initial inoculum density, as strains of P. polymyxa with a strong antimicrobial activity (including P. polymyxa GBR-462) showed the antimicrobial activity against P. capsici and could form biofilm only when they were applied at the higher initial inoculums, 10⁸ cfu/ml. No inhibitory effect was noted on the mycelial growth and zoospore germination of the pathogen when applied at the lower inoculum density of 10⁶ cfu/ml of P. polymyxa GBR-462. However, sporangium formation and zoospore release was significantly inhibited at the lower inoculum density. Also light and electron microscopy revealed the structures of sporangia aberrant with no or few healthy nuclei, indicating sporangium and zoospore formation inhibited at the lower inoculum density. Application of P. polymyxa GBR-462 into potted soil suppressed disease progression as well as disease severity; disease severity was reduced by 30% as compared to untreated pots, suggesting P. polymyxa GBR-462 could be a potential biocontrol agent against Phytopthora capsici .     

培菲康双歧三联活菌制剂中三株菌对抗菌药物敏感性检测

目的：检测培菲康活菌制剂中双歧杆菌,嗜酸乳杆菌与粪链球菌三株菌对30种抗菌药物的敏感性。方法：用纸片琼脂扩散法（disc agar diffusion,即Kirby-Bauer法）和微量稀释法（microdilution bro9th method)进行检测。结果：这三株菌对大多数抗菌药物敏感,对少数抗菌药物耐受。与文献报导相似,这三株菌与相应的国际参考菌株及临床菌株比较,没有发现明显耐药变异。结论：为避免抗菌药物对双歧三联活菌制剂功效的影响。每天两药宜分开时间服用。服用双歧三联活菌不会引起耐药性的扩散。     

Antimicrobial and membranolytic activities of anti-burn drug fenozan]

Fenozan, an anti-burn preparation, was shown to have antimicrobial activity against freshly isolated clinical strains of Staphylococcus aureus and Streptococcus faecalis, as well as against collection strains of the other gram-positive bacteria. The antimicrobial action of the preparation was possibly due to impairment of permeability of the cytoplasmic membranes in the sensitive bacterial cells and their liberation of intracellular low molecular weight compounds to the environment. The membranolytic and minimum inhibitory concentrations of fenozan with respect to the sensitive bacterial cells were one order of magnitude lower than the concentration stabilizing the membranes of animal cells in the treatment of burns. Combination of the antioxidant and antimicrobial properties in fenozan was likely to provide its satisfactory therapeutic effect in the treatment of burn wounds.     

Escherichia coli O157 and non-O157 isolates are more susceptible to L-lactate than to D-lactate

The antimicrobial effect of L-lactate was much greater than that of D-lactate over a range of concentrations for Escherichia coli O157 and non-O157 strains. Despite this, the intracellular pHs and membrane potentials of L-lactate- and D-lactate-treated cells were similar, suggesting that these factors are not involved in the antimicrobial action of L-lactate.     

Targeting Imperfect Vaccines against Drug-Resistance Determinants: A Strategy for Countering the Rise of Drug Resistance

The growing prevalence of antimicrobial resistance in major pathogens is outpacing discovery of new antimicrobial classes. Vaccines mitigate the effect of antimicrobial resistance by reducing the need for treatment, but vaccines for many drug-resistant pathogens remain undiscovered or have limited efficacy, in part because some vaccines selectively favor pathogen strains that escape vaccine-induced immunity. A strain with even a modest advantage in vaccinated hosts can have high fitness in a population with high vaccine coverage, which can offset a strong selection pressure such as antimicrobial use that occurs in a small fraction of hosts. We propose a strategy to target vaccines against drug-resistant pathogens, by using resistance-conferring proteins as antigens in multicomponent vaccines. Resistance determinants may be weakly immunogenic, offering only modest specific protection against resistant strains. Therefore, we assess here how varying the specific efficacy of the vaccine against resistant strains would affect the proportion of drug-resistant vs. –sensitive strains population-wide for three pathogens – Streptococcus pneumoniae, Staphylococcus aureus, and influenza virus – in which drug resistance is a problem. Notably, if such vaccines confer even slightly higher protection (additional efficacy between 1% and 8%) against resistant variants than sensitive ones, they may be an effective tool in controlling the rise of resistant strains, given current levels of use for many antimicrobial agents. We show that the population-wide impact of such vaccines depends on the additional effect on resistant strains and on the overall effect (against all strains). Resistance-conferring accessory gene products or resistant alleles of essential genes could be valuable as components of vaccines even if their specific protective effect is weak.     

Study of antimicrobial activity amongLactobacillus helveticus strains using three different assays

The antimicrobial activity of 18Lactobacillus helveticus strains as well as one control strain, the bacteriocin producingLactobacillus helveticus 481, was tested with three different inhibition assays. FourLactobacillus helveticus strains had antimicrobial activity against seven otherLactobacillus helveticus strains and twoLactobacillus delbrueckil strains while the remaining sevenLactobacillus helveticus strains were indifferent. Inhibition was also observed againstLactococcus andLeuconostoc species, due to production of organic acids or hydrogen peroxide. The strains with the highest antimicrobial activity produced a heat sensitive proteinacious bacteriocin with a narrow species activity spectrum against only thermophilicLactobacillus strains.     

Antimicrobial evaluation of coumarins and flavonoids from the stems of Daphne gnidium L.

The antimicrobial activity of stems methanol extract from Daphne gnidium L. collected from Sardinia (Italy) was evaluated against 6 strains of standard and clinical isolated gram (+/-) bacteria. The antimicrobial effect on two strains of fungi was also tested. The extract in toto exhibited antibacterial activity against Bacillus lentus and Escherichia coli, but was inactive against fungi. Four coumarins (daphnetin, daphnin, acetylumbelliferone, daphnoretin) and seven flavonoids (luteolin, orientin, isoorientin, apigenin-7-O-glucoside, genkwanin, 5-O-beta-D-primeverosyl genkwanine, 2,5,7,4'-tetrahydroxyisoflavanol) present in the plant extract were also investigated against the same strains of bacteria and fungi assayed for the crude extract. The most active compounds were daphnetin, genkwanin, and 2,5,7,4'-tetrahydroxyisoflavanol.     

Effects of antimicrobial treatment on fiberglass-acrylic filters

AIMS: The aims of the present study were to: (i) analyse a group of antimicrobial agents and to select the most active against test microbial strains; (ii) test the effect of the antimicrobial treatment on air filters in order to reduce microbial colonization. METHODS AND RESULTS: Different kinds of antimicrobial agents were analysed to assess their compatibility with the production process of air filter media. The minimal inhibitory concentration for each antimicrobial agent was determined against a defined list of microbial strains, and an antimicrobial activity assay of filter prototypes was developed to determine the most active agent among the compatible antimicrobials. Then, the most active was chosen and added directly to the filter during the production process. The microbial colonization of treated and untreated filter media was assessed at different working times for different incubation times by stereomicroscope and scanning electron microscope analysis. Some of the antimicrobial agents analysed were more active against microbial test strains and compatible with the production process of the filter media. Filter sections analysis of treated filter media showed a significantly lower microbial colonization than those untreated, a reduction of species both in density and varieties and of the presence of bacteria and fungal hyphae with reproductive structures. CONCLUSIONS: This study demonstrated the ability of antimicrobial treatments to inhibit the growth of micro-organisms in filter media and subsequently to increase indoor air quality (IAQ), highlighting the value of adding antimicrobials to filter media. SIGNIFICANCE AND IMPACT OF THE STUDY: To make a contribution to solving the problem of microbial contamination of air filters, by demonstrating the efficacy of incorporating antimicrobial agents in the filter media to improve IAQ and health.     

银杏内生菌的分离及其抑菌活性筛选

从健康的银杏（Ginkgo biloba）茎和叶片中分离内生菌,结果从银杏叶和茎上共分离到内生真菌20株,其中9株来自银杏茎部,11株来自银杏叶部;内生放线菌23株,其中15株来自于银杏茎部,8株来自于银杏叶部;内生细菌15株,其中8株来自于银杏茎部,7株来自于银杏叶部。以金黄色葡萄球菌（Staphylococcus aureus）、枯草芽胞杆菌（Bacillus subtilis）、大肠埃希菌（Escherichia coli）、黑曲霉（Aspergillus niger）、酿酒酵母（Saccharomyces cerevisiae）作为指示菌,采用双层平板法对内生菌进行抑菌活性筛选,结果表明：20株内生真菌中有12株出现了抑菌活性,有抑菌活性菌株的比例为60.0%;23株内生放线菌中,仅3株出现了抑菌活性,有抑菌活性菌株的比例为13.0%;15株内生细菌中,有4株出现了抑菌活性,有抑菌活性菌株的比例为26.7%。     

Prevalence of the genes encoding propionicin T1 and protease-activated antimicrobial peptide and their expression in classical propionibacteria

The purpose of this study was to investigate the frequency of production of the bacteriocin propionicin T1 and the protease-activated antimicrobial peptide (PAMP) and their corresponding genes in 64 isolates of classical propionibacteria. This study revealed that these genes are widespread in Propionibacterium jensenii and Propionibacterium thoenii but absent from the remaining species of classical propionibacteria that were studied. The pro-PAMP-encoding gene (pamA) was found in 63% of the P. jensenii strains and 61% of the P. thoenii strains, and all of these strains displayed PAMP activity. The propionicin T1-encoding gene (pctA) was present in 89% of the P. thoenii strains and 54% of the P. jensenii strains. All P. thoenii strains containing the pctA gene exhibited antimicrobial activity corresponding to propionicin T1 activity, whereas only 38% of the pctA-containing P. jensenii strains displayed this activity. Sequencing of the pctA genes revealed the existence of two allelic variants that differed in a single nucleotide in six strains of P. jensenii; in these strains the glycine at position 55 of propionicin T1 was replaced by an aspartate residue (A variant). No strains harboring the A variant showed any antimicrobial activity against propionicin T1-sensitive bacteria. An open reading frame (orf2) located immediately downstream from the pctA gene was absent in three strains containing the G variant of propionicin T1. Two of these strains showed low antimicrobial activity, while the third strain showed no antimicrobial activity at all. The protein encoded by orf2 showed strong homology to ABC transporters, and it has been proposed previously that this protein is involved in the producer immunity against propionicin T1. The limited antimicrobial activity exhibited by the strains lacking orf2 further suggests that this putative ABC transporter plays an important role in propionicin T1 activity.     

Salt resistance and synergistic effect with vancomycin of alpha-helical antimicrobial peptide P18.

P18 (KWKLFKKIPKFLHLAKKF-NH(2)) is an alpha-helical antimicrobial peptide designed from a cecropin A-magainin 2 hybrid. In this study, P18 was found to show strong antimicrobial activity against several antibiotic-resistant bacterial and fungal strains. Both the salt resistance on antimicrobial activity and the synergistic effect with clinically used antibiotic agents are critical factors in developing effective peptide antibiotic drugs. For this reason, we investigated the salt resistance of P18 to antagonism by NaCl, CaCl(2), and MgCl(2) on antimicrobial activity and the synergistic effect of P18 with vancomycin against vancomycin-resistant Enterococcus faecium (VREF). Compared to magainin 2, P18 showed strong resistance on antimicrobial activity against bacterial strains and C. albicans under high NaCl concentrations of 100-200 mM. In addition, P18 displayed much greater salt resistance on antibacterial activity against Gram-negative bacteria at the physiological or elevated concentrations of CaCl(2) and MgCl(2) than magainin 2. Furthermore, the combination study revealed that P18 has a relatively effective synergistic effect with vancomycin against VREF. Thus, these results support that P18 may prove to be a salt-resistant antibiotic peptide potentially useful in the treatment of cystic fibrosis patients as well as a valuable adjuvant for antimicrobial chemotherapy.     

β-Lapachone activity in synergy with conventional antimicrobials against methicillin resistant Staphylococcus aureus strains

The aim of this study was to evaluate the antimicrobial activity of lapachol, α-lapachone, β-lapachone and six antimicrobials (ampicillin, amoxicillin/clavulanic acid, cefoxitin, gentamicin, ciprofloxacin and meropenem) against twelve strains of Staphylococcus aureus from which resistance phenotypes were previously determined by the disk diffusion method. Five S. aureus strains (LFBM 01, LFBM 26, LFBM 28, LFBM 31 and LFBM 33) showed resistance to all antimicrobial agents tested and were selected for the study of the interaction between β-lapachone and antimicrobial agents, busing checkerboard method. The criteria used to evaluate the synergistic activity were defined by the Fractional Inhibitory Concentration Index (FICI). Among the naphthoquinones, β-lapachone was the most effective against S. aureus strains. FICI values ranged from 0.07 to 0.5, suggesting a synergistic interaction against multidrug resistant S. aureus (MRSA) strains. An additive effect was observed with the combination β-lapachone/ciprofloxacin against the LFBM 33 strain. The combination of β-lapachone with cefoxitin showed no added benefit against LFBM 31 and LFBM 33 strains. This study demonstrated that, in general, β-lapachone combined with beta lactams antimicrobials, fluoroquinolones and carbapenems acts synergistically inhibiting MRSA strains.     

Development of an antimicrobial formulation for control of specimen-related contamination in phage-based diagnostic testing for tuberculosis

AIMS: To develop and evaluate an antimicrobial supplement for use with phage-based tests for rapid detection of drug resistance of tuberculosis (TB). METHODS AND RESULTS: An antimicrobial formulation containing nystatin, oxacillin and aztreonam (NOA) (final concentrations of 50,000 IU l(-1), 2 mg l(-1), and 30 mg l(-1) respectively) was developed. This formulation was tested for its influence on detection of a number of Mycobacterium tuberculosis (MTB) strains using the phage amplification (FASTPlaque) assay. Addition of the supplement did not lead to significant reduction in assay sensitivity. Antimicrobial efficacy was assessed with a range of Gram-positive and -negative organisms. The NOA supplement had a broad antimicrobial effect. The supplement was tested for its effect on growth of MTB culture, and on determination of rifampicin resistance using the phage-based methodology (FASTPlaque-Response). NOA did not significantly affect the growth of a range of rifampicin susceptible and resistant MTB strains, nor did it have an adverse effect on the number of interpretable results, nor the ability to discriminate between rifampicin susceptibility and resistance. CONCLUSION, SIGNIFICANCE AND IMPACT OF STUDY: Use of NOA antimicrobial supplement with rapid phage-based tests for TB will increase the proportion of interpretable results obtained, and enable their wider implementation in disease-endemic countries by improved control of specimen-related contamination.     

Antimicrobial effect of fermented Ghanaian maize dough

Unhygienic conditions of a typical rural community in a developing country were simulated in the laboratory by inoculating fermented maize dough porridge with Shigella flexneri and enterotoxigenic Escherichia coli (ETEC). The antimicrobial effects of the different processes involved in the preparation of fermented maize dough porridge were assessed. The soaking process reduced the pH but no antimicrobial effect against shigella and ETEC was noted. Unfermented maize dough did not inhibit any of the test strains. When the fermentation process had become established, half of the strains tested were inhibited by the fermented maize dough when examined 8 h after inoculation. Cooking the fermented maize dough into porridge reduced the antimicrobial effect but there was still significant inhibition of pathogens. This suggests that the antimicrobial effect of fermented maize dough is not due to pH per se. Fermentation of maize dough appears to be a useful strategy for reducing contamination of weaning foods by Sh. flexneri and ETEC. The possible nature of the antimicrobial agent(s) produced during the fermentation of maize dough is discussed.     

Antimicrobial effect of fermented Ghanaian maize dough

Unhygienic conditions of a typical rural community in a developing country were simulated in the laboratory by inoculating fermented maize dough porridge with Shigella flexneri and enterotoxigenic Escherichia coli (ETEC). The antimicrobial effects of the different processes involved in the preparation of fermented maize dough porridge were assessed. The soaking process reduced the pH but no antimicrobial effect against shigella and ETEC was noted. Unfermented maize dough did not inhibit any of the test strains. When the fermentation process had become established, half of the strains tested were inhibited by the fermented maize dough when examined 8 h after inoculation. Cooking the fermented maize dough into porridge reduced the antimicrobial effect but there was still significant inhibition of pathogens. This suggests that the antimicrobial effect of fermented maize dough is not due to pH per se. Fermentation of maize dough appears to be a useful strategy for reducing contamination of weaning foods by Sh. flexneri and ETEC. The possible nature of the antimicrobial agent(s) produced during the fermentation of maize dough is discussed.     

渤海湾仿刺参养殖圈内贝莱斯芽胞杆菌B-VE的分离鉴定及其产抗菌脂肽的研究

为分离筛选具有广谱抑菌作用的细菌并研究其抗菌物质,实验利用琼脂扩散法和牛津杯法,以溶壁微球菌、金黄色葡萄球菌、铜绿假单胞杆菌、大肠埃希菌为指示菌,从渤海湾大连海域仿刺参养殖圈中分离筛选出9株活性菌,2株具有广谱抑菌效果,其中1株抑菌效果最强.通过形态学和分子生物学检测分析,鉴定该菌株为贝莱斯芽胞杆菌(Bacillus ...     

In vitro activities of antibiotics and antimicrobial peptides against the plant pathogenic bacterium Xylella fastidiosa

AIMS: The objective of this study was to evaluate the effectiveness of antibiotics and antimicrobial peptides against 10 strains of Xylella fastidiosa. METHODS AND RESULTS: The minimal inhibitory concentration (MIC) of 12 antibiotics and 18 antimicrobial peptides were determined by agar dilution tests and growth inhibition assays. Antibiotics with the lowest MIC for X. fastidiosa strains were gentamicin, tetracycline, ampicillin, kanamycin, and novobiocin, chloramphenicol, and rifampin. Plate growth inhibition assays showed that four of the antimicrobial peptides (Magainin 2, Indolicidin, PGQ, and Dermaseptin) were toxic to all X. fastidiosa strains. CONCLUSION: All X. fastidiosa strains were sensitive to several groups of antibiotics, and minor differences in sensitivity to several antimicrobial peptides were observed among strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that antibiotics and antimicrobial peptides have some activity against the pathogen, X. fastidiosa and may have application in protecting plants from developing Pierce's disease.