Effect of Inoculum Size on In Vitro Susceptibility Testing with Lincomycin

There is disagreement in the literature as to whether lincomycin is primarily a bacteriostatic or a bactericidal agent against gram-positive cocci and also regarding the levels of activity of this agent against susceptible microorganisms. These questions were examined in a study of the effect of inoculum size on the results of tube dilution susceptibility determinations with lincomycin against 49 clinical isolates of Staphylococcus aureus and 25 strains of streptococci and pneumococci. Lincomycin was both highly active and bactericidal when tested against 40 strains of S. aureus with inocula containing a maximum of 10(4) cells per ml [median minimal inhibitory concentration (MIC), 0.78 mug/ml; median minimal bactericidal concentration (MBC), 1.56 mug/ml]. With inocula of 10(5) cells per ml, lincomycin was primarily bacteriostatic (median MIC, 1.56 mug/ml; median MBC, 12.5 mug/ml). There were further decreases in inhibitory levels and significant losses of bactericidal activity when inocula containing more than 10(7) cells were tested (median MIC, 3.13 mug/ml; median MBC > 100 mug/ml). Similar measurements with streptococci and pneumococci revealed a lesser effect of inoculum size. The mean MBC value for alpha-hemolytic streptococci increased from 0.40 to 1.05 mug/ml with an increase in inocula from 10(4) to 10(6) cells per ml, but without a marked increase in MIC values. Similar results were obtained for beta-hemolytic streptococci and pneumococci.     

抗菌肽体外对结核杆菌的作用初探

将结核杆菌接种于含抗菌肽的苏通氏培养基中 ,在接种后不同时间内取样接种于罗氏培养基上 ,观察结核杆菌在罗氏培养基上的生长情况 ,初步探讨了抗菌肽 (CecropinB)对结核杆菌标准株H3 7RV的作用。结果显示 ,接种 10天后在罗氏培养基上结核杆菌对抗菌肽敏感。说明在体外培养中 ,抗菌肽有一定的抗结核杆菌的作用     

Comparative Study of Agar Diffusion Test and the NCCLS Macrobroth Method for In Vitro Susceptibility Testing of Candida spp.

We performed a prospective double-blind study to evaluate the correlation between inhibition zones obtained by a disk-diffusion test, using Neo-sensitabs of fluconazole (Rosco Diagnostica), and the MICs generated by the NCCLS macrobroth dilution assay. Eighty clinical isolates, representing 5 of the clinically relevant species of Candida, were tested simultaneously by both methods. A clear inverse correlation was found between the results obtained by both tests (r = −0.69). In addition, there was high degree of agreement between methods in the identification of susceptible isolates. However, the resistance definition by disk-diffusion test had a positive predictive value of only 17%. Our data support the hypothesis that Rosco Fluconazole Neo-sensitabs have potential as a screening test for the identification of Candida isolates susceptible to fluconazole. Resistant isolates should be further investigated by standardized broth procedures. This revised version was published online in June 2006 with corrections to the Cover Date.     

The Relationship Between Agar Thickness and Antimicrobial Susceptibility Testing

Antimicrobial susceptibility testing can be done using solid or liquid-based medium. Solid-based assays are easy and inexpensive; they are limited by not being as quantitative as liquid-based assays. Agar depth can influence the accuracy of plate-based assays and it is assumed the basis of this effect is antimicrobial agent diffusion. We tested this assumption by using ETEST^® to quantitate the relationship between agar depth and minimum inhibitory concentration and zone of inhibition.     

Determination of Untreated Whole-Milk Effects on In Vitro Antibacterial Activity

The effect of fresh whole milk without pasteurization or other pretreatment on in vitro antibacterial activity of selected compounds was determined in broth dilution. The milk was collected by hand directly from dairy goats, or by syringe or cannula from bovine quarters showing low bacterial counts. Antibacterial activity was determined in 50% (v/v) milk-broth medium against sensitive mastitis-etiologic strains of Streptococcus agalactiae and Staphylococcus aureus. The indicator salt 2,3,5-triphenyltetrazolium chloride was incorporated in the milk broth medium to determine inoculum growth. Contaminant interference was circumvented through early as well as late readings and comparisons with uninoculated control tubes, with and without the test compounds. Application of the method with more than 75 compounds, including nitrofurans, antibiotics, and other chemicals uncovered marked degrees of milk interference. The method warrants routine use among preliminary screens to relate in vitro with in vivo observations of antimicrobial activity. Similar procedures may be used with serum, skim milk, or mastitis-milk media for separating effects due to protein, lipid, or other elements in product evaluation.     

大蒜中生理活性物质对致病细菌的体外抑制作用

目的：从鲜蒜中提取蒜氨酸、蒜氨酸酶和蒜素,测定了蒜氨酸、蒜氨酸酶、蒜氨酸＋蒜氨酸酶、蒜素和蒜汁对10种致病细菌的抑菌作用及最低抑菌浓度。方法：采用平板打孔法和液体2倍稀释法。结果：蒜氨酸酶、蒜氨酸＋蒜氨酸酶、蒜素和蒜汁对金黄色葡萄球菌、大肠杆菌、鲍曼不动杆菌等10种菌株的抑菌环直径均大于7mm。蒜氨酸＋蒜氨酸酶对鲍曼不动杆菌的抑菌作用是50mg／mL氨苄青霉素的1．5倍。蒜氨酸＋蒜氨酸酶对金黄色葡萄球菌、类产碱假单胞菌和福氏志贺氏菌的MIC为0．19mg/mL。结论：蒜氨酸酶、蒜氨酸＋蒜氨酸酶、蒜素和蒜汁在体外具有明显的抑菌作用。     

青天葵体外抑菌活性研究

目的:研究青天葵水提取液、醇提取液和水醇提取液对金黄色葡萄球菌、大肠杆菌、白色念珠菌、伤寒沙门菌、绿脓杆菌、黑曲霉菌6种菌株的抑菌效果。方法:制备青天葵3种提取液,采用试管两倍稀释法测定3种提取液对金黄色葡萄球菌等6种菌株的最低抑菌浓度(MIC)。结果:青天葵水提取液对伤寒沙门菌的抑菌作用较强(MIC为12.5%),对金黄色葡萄球菌、大肠杆菌、绿脓杆菌和黑曲霉菌的抑菌作用较弱(MIC为50%);醇提取液对伤寒沙门菌和绿脓杆菌都有很强的抑菌作用(MIC为6.25%),对金黄色葡萄球菌也有较强的抑菌活性(MIC为12.5%),对大肠杆菌的抑菌作用较弱(MIC为50%);水醇提取液的抑菌活性与醇提取液相当。结论:青天葵对金黄色葡萄球菌等6种菌株表现出不同程度的抑制活性。     

Synthesis and In Vitro Antibacterial Activity of New C-3-Modified Carbapenems

Russian Journal of Bioorganic Chemistry - New C-3 modified carbapenems have been synthesized by the AdNE-substitution of the enol phosphate group of 4-nitrobenzyl...     

四氮唑类化合物的合成及体外抑菌活性研究

目的:利用点击化学反应合成四氮唑类化合物,寻找温和的反应条件。对合成的四氮唑类化合物进行体外抑菌活性研究,以期发现抑菌活性化合物并初步研究其构效关系。方法:以氰基类化合物和叠氮钠为原料,溴化锌为催化剂,通过点击化学反应合成一类四氮唑结构化合物并研究其合成工艺条件的优化。利用微量二倍稀释法对合成的四氮唑类化合物进行体外抑菌活性测试。结果:利用所得到的优化点击化学反应条件合成了11个四氮唑类化合物,并发现化合物2h具有广谱抑菌活性。结论:优化后的点击化学反应条件温和,后处理方便,产率较高。抑菌活性和构效关系研究为后续的四氮唑类化合物抑菌活性研究工作提供了一定的基础。     

Chlorhexidine Salt-Loaded Polyurethane Orthodontic Chains: In Vitro Release and Antibacterial Activity Studies

The widespread use of indwelling medical devices has enormously increased the interest in materials incorporating antibiotics and antimicrobial agents as a means to prevent dangerous device-related infections. Recently, chlorhexidine-loaded polyurethane has been proposed as a material suitable for the production of devices which are able to resist microbial contamination. The aim of the present study was to characterize the in vitro release of chlorhexidine from new polymeric orthodontic chains realized with polyurethane loaded with two different chlorhexidine salts: chlorhexidine diacetate or chlorhexidine digluconate. The orthodontic chains constituted of three layers: a middle polyurethane layer loaded with chlorhexidine salt inserted between two layers of unloaded polymer. In vitro release of chlorhexidine diacetate and digluconate from orthodontic chains loaded with 10% or 20% (w/w) chlorhexidine salt was sustained for 42 days and followed Fickian diffusion. The drug diffusion through the polyurethane was found to be dependent not only on chlorhexidine loading, but also on the type of chlorhexidine salt. The antibacterial activity of 0.2% (w/w) chlorhexidine diacetate-loaded orthodontic chain was successfully tested towards clinically isolated biofilm forming ica-positive Staphylococcus epidermidis via agar diffusion test. In conclusion, the chlorhexidine salt-loaded chains could provide an innovative approach in the prevention of oral infections related to the use of orthodontic devices.KEY WORDS: antibacterial activity, cariogenic treatment, chlorhexidine, in vitro release, orthodontic chains     

In Vitro Antibacterial Activity of Cysteine Protease Inhibitor from Kiwifruit (Actinidia deliciosa)

The need for replacing traditional pesticides with alternative agents for the management of agricultural pathogens is rising worldwide. In this study, a cysteine proteinase inhibitor (CPI), 11 kDa in size, was purified from green kiwifruit to homogeneity. We examined the growth inhibition of three plant pathogenic Gram-negative bacterial strains by kiwi CPI and attempted to elucidate the potential mechanism of the growth inhibition. CPI influenced the growth of phytopathogenic bacteria Agrobacterium tumefaciens (76.2 % growth inhibition using 15 μM CPI), Burkholderia cepacia (75.6 % growth inhibition) and, to a lesser extent, Erwinia carotovora (44.4 % growth inhibition) by inhibiting proteinases that are excreted by these bacteria. Identification and characterization of natural plant defense molecules is the first step toward creation of improved methods for pest control based on naturally occurring molecules.     

In Vitro Susceptibility of Atypical Mycobacteria To Rifampin

Atypical mycobacteria (209 strains) were examined for susceptibility to rifampin by the proportion method by using Middlebrook 7H-10 agar. All strains of Mycobacterium kansasii and tap-water scotochromogens were inhibited by 0.25 to 1 μg of the drug per ml. Seventy-six per cent of M. scrofulaceum and 61% of M. intracellulare strains were susceptible to 4 μg/ml or less; 5% of the former and 8% of the latter were resistant to 16 μg/ml. All strains of M. gastri and M. triviale and most strains of M. terrae were sensitive to 1 to 4 μg/ml. Two strains of M. borstelense were both inhibited by 8 μg/ml. Nearly all strains of M. fortuitum were resistant to the drug. The results of this study suggest that rifampin may be a valuable agent for the treatment of many atypical mycobacterial infections.     

A Reference Broth Microdilution Method for Dalbavancin In Vitro Susceptibility Testing of Bacteria that Grow Aerobically

Antimicrobial susceptibility testing (AST) is performed to assess the in vitro activity of antimicrobial agents against various bacteria. The AST results, which are expressed as minimum inhibitory concentrations (MICs) are used in research for antimicrobial development and monitoring of resistance development and in the clinical setting for antimicrobial therapy guidance. Dalbavancin is a semi-synthetic lipoglycopeptide antimicrobial agent that was approved in May 2014 by the Food and Drug Administration (FDA) for the treatment of acute bacterial skin and skin structure infections caused by Gram-positive organisms. The advantage of dalbavancin over current anti-staphylococcal therapies is its long half-life, which allows for once-weekly dosing. Dalbavancin has activity against Staphylococcus aureus (including both methicillin-susceptible S. aureus [MSSA] and methicillin-resistant S. aureus [MRSA]), coagulase-negative staphylococci, Streptococcus pneumoniae, Streptococcus anginosus group, β-hemolytic streptococci and vancomycin susceptible enterococci. Similar to other recent lipoglycopeptide agents, optimization of CLSI and ISO broth susceptibility test methods includes the use of dimethyl sulfoxide (DMSO) as a solvent when preparing stock solutions and polysorbate 80 (P80) to alleviate adherence of the agent to plastic. Prior to the clinical studies and during the initial development of dalbavancin, susceptibility studies were not performed with the use of P-80 and MIC results tended to be 2-4 fold higher and similarly higher MIC results were obtained with the agar dilution susceptibility method. Dalbavancin was first included in CLSI broth microdilution methodology tables in 2005 and amended in 2006 to clarify use of DMSO and P-80. The broth microdilution (BMD) procedure shown here is specific to dalbavancin and is in accordance with the CLSI and ISO methods, with step-by-step detail and focus on the critical steps added for clarity.     

In Vitro Susceptibility of Selected Bacteria to Cefaclor

Cefaclor is an orally absorbed cephalosporin antibiotic chemically and pharmacologically similar to cephalexin. It appears to be more active than cephalexin against susceptible strains. The in vitro sensitivity of 230 clinical bacterial isolates to cefaclor was studied. Most isolates of S. aureus, K. pneumoniae, E. coli, and indole negative Proteus species were inhibited at clinically attainable serum and urine concentrations. Like cephalexin, cefaclor was less active against isolates of Enterobacter species, indole positive Proteus species and enterococci although many of these isolates were inhibited at concentrations achievable in urine.     

In Vitro Susceptibility of Brucella to Various Antibiotics

A series of 27 strains of six species of Brucella was tested for susceptibility in vitro to a representative cross section of antibiotics in current use. The activity against each species was plotted, with the cumulative per cent of strains inhibited indicated for each concentration. As a class, the tetracycline antibiotics were the most effective. Erythromycin, gentamicin, streptomycin, and kanamycin, as well as rifampin, were quite active. The penicillin-cephalosporin group, with the exception of ampicillin, was comparatively ineffective, as were the polypeptides and the miscellaneous group of chloramphenicol, lincomycin, cycloserine, and sulfadiazine. Species differences were noticeable, with some strains of B. canis being considerably more resistant to streptomycin and the tetracyclines than B. suis and B. abortus. B. melitensis, B. ovis, and B. neotomae were intermediate in antibiotic susceptibility.     

In Vitro Antiviral Activity of Mycophenolic Acid and Its Reversal by Guanine-Type Compounds

With the agar diffusion test and BS-C-1 cells, mycophenolic acid was found to give a straight-line dose-response activity in inhibiting the cytopathic effects of vaccinia, herpes simplex, and measles viruses. Plaque tests have shown 100% reduction of virus plaques by mycophenolic acid over drug ranges of 10 to 50 mug/ml and virus input as high as 6,000 plaque-forming units (PFU) per flask. Back titration studies with measles virus inhibited by mycophenolic acid have indicated that extracellular virus titers were reduced by approximately 3 logs(10) and total virus was reduced by 1 log(10). The agar diffusion test system lends itself readily to drug reversal studies. Mycophenolic acid incorporated into agar at 10 mug/ml gave 100% protection to virus-infected cells. Filter paper discs impregnated with selected chemical agents at concentrations of 1,000 mug/ml (20 mug per filter paper disc) were placed on the agar surface. Reversal of the antiviral activity of mycophenolic acid was indicated by virus breakthrough in those cells in close proximity to the filter paper disc. Chemicals showing the best reversal of the antiviral activity of mycophenolic acid were guanine, guanosine, guanylic acid, deoxyguanylic acid, and 2,6-diaminopurine. The reversal of antiviral activity was confirmed by titrations of virus produced with various amounts of both mycophenolic acid and guanine present and by isotope tracer methods with uptakes of labeled uridine, guanine, leucine, and thymidine in treated and nontreated, infected and noninfected cells as parameters. All antiviral effects of mycophenolic acid at 10 mug/ml could be reversed to the range shown by untreated controls by the addition of 10 mug/ml of those chemicals exhibiting reversal activity.     

斗米虫蛋白体外抗肿瘤活性及其对小鼠巨噬细胞免疫调节作用

该文主要为了研究斗米虫蛋白的体外抗肿瘤活性及其免疫调节作用。提取斗米虫总蛋白,逐级盐析分为3个部分,透析除盐后得到不同蛋白部位,并采用SDS-PAGE检测斗米虫不同蛋白部位的分子量;利用MTT法、流式细胞术等方法研究斗米虫蛋白对人胃癌细胞MFC和小鼠乳腺癌细胞4T1增殖、迁移和凋亡的作用。MTT法研究斗米虫蛋白对小鼠单核巨噬细胞RAW264.7和人脐静脉内皮细胞HUEVC增殖的影响;荧光微球吞噬实验检测斗米虫蛋白对RAW264.7细胞吞噬能力的影响;Griess法检测对RAW264.7细胞释放NO能力的影响;ELISA法检测对RAW264.7细胞的IL-6、TNF-α和IL-1β分泌量;RT-PCR法检测不同浓度的斗米虫蛋白作用后RAW264.7细胞中TNF-α、IL-1、TLR4、MIR-7、IFN-γ、TRL-4、IL-6以及4T1细胞中MMP2、MMP9、STAT3、c-Myc和Sdf1 mRNA水平变化。结果显示,斗米虫蛋白主要分子量集中于63 kDa,斗米虫蛋白对人胃癌细胞MFC及小鼠乳腺癌细胞4T1的增殖表现出较好抑制作用,并呈现出一定剂量依赖关系(P<0.05),对HUEVC细胞没有细胞毒性,对RAW264.7细胞表现出较好的促进增殖的作用(P<0.05)。斗米虫蛋白实验组与正常组细胞相比可以抑制4T1细胞的迁移(P<0.01),可诱导MFC和4T1细胞凋亡(P<0.05);斗米虫蛋白能够提高RAW264.7细胞的吞噬活性、NO释放量、TNF-α、1L-1β和IL-6分泌量及TNF-α、IL-1、TLR4、MIR-7、IFN-γ和IL-6细胞因子的mRNA水平以及能显著下调4T1细胞中MMP2、MMP9、STAT3、c-Myc和Sdf1 mRNA水平(P<0.05,P<0.01)。由此推论,斗米虫蛋白具有较好的体外抗肿瘤活性并且具有潜在的免疫调节作用。