Antimicrobial activity of organic extracts isolated from Aplysina fistularis (Demospongiae: Aplysinidae)

Organic extracts of the sponge Aplysina fistularis (Pallas 1766) were tested for antimicrobial activity against Gram positive bacteria (Staphylococcus aureus) and Gram negative bacteria (Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa). The minimal inhibitory concentration (MIC) and toxic activity of extract were determined. Susceptibility trials of organic fractions obtained by VLC: Hexane, EtOAc and CHCl3 showed that EtOAc fraction has antibacterial activity against E. coli, while CHCl3 fraction inhibited E. coli and S. aureus growth. The later refractioning of EtOAc fraction and the biodirected assays showed that fractions F12 and F13 of EtOAc/Hex and EtOAc F14 were bioactive against Gram positive and Gram negative bacteria. Only EtOAc/MeOH Sf2 from subfractionig of EtOAc F14 produced inhibition for E. coli and S. aureus. In Sf2 EtOAc/MeOH, MIC was moderate for S. aureus (MIC > 256 g/ml). F4 CHCl3/MeOH produced a high inhibition in S. aureus (MIC = 0.125 g/ml) and for E. coli (MIC > 16 g/ml). F10 CHCl3/MeOH showed a moderate activity against S. aureus (MIC > 128 g/ml) and low activity against E. coli (MIC = 512 g/ml). F10 CHCL3/MeOH did no present toxic activity against Artemia salina. The fractiorts F4 CHCL3/MeOH and Sf2 EtOAc/MeOH were toxic for this organism when the concentration was higher than 100 microg/ml. LC50 in both cases was 548.4 and 243.4 microg/ml respectively. Secondary metabolites of medium polarity obtained from A. fistularis have a wide spectrum of anti bacterial activity. Toxicity analysis suggests that only F10 CHCL3/MeOH has potential as an antimicrobial agent for clinical use.     

黑大蒜提取物联合抗生素对金黄色葡萄球菌和肠埃希菌的体外抗菌作用研究

评价黑大蒜提取物分别与头孢唑林或庆大霉素联合应用,对金黄色葡萄球菌和大肠埃希菌的体外抗菌效应。采用液体稀释法分别测定黑大蒜提取物对金黄色葡萄球菌和大肠埃希菌的最低抑菌浓度（MIC）。采用棋盘法设计,微量肉汤稀释法测定黑大蒜提取物联合头孢唑林或庆大霉素对金黄色葡萄球菌和大肠埃希菌的MIC,并计算部分抑菌浓度（FIC指数）。测定黑大蒜提取物对金黄色葡萄球菌和大肠埃希菌的时间-杀菌曲线。黑大蒜提取物对金黄色葡萄球菌的MIC为256μg/mL,黑大蒜提取物对大肠埃希菌的MIC为256μg/mL。时间-杀菌曲线结果显示黑大蒜提取物对金黄色葡萄球菌和大肠埃希菌的抑菌作用呈现较强的浓度依赖性。黑大蒜提取物联合头孢唑林后对金黄色葡萄球菌的FIC指数为0.75;黑大蒜提取物联合庆大霉素后对大肠埃希菌的FIC指数为0.5。黑大蒜提取物与头孢唑林或庆大霉素联合用药,可明显降低抗生素对金黄色葡萄球菌和大肠埃希菌的MIC,表现为相加和协同效应。     

In vitro antibacterial activity of laboratory grown culture of Spirulina platensis

The hexane, ethyl acetate, dichloromethane, methanol extracts and spent media (extracellular substances) were tested in vitro for their antibacterial activity for which one Gram-positive bacterium (Staphylococcus aureus) and four Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, and Klebsiella pneumoniae) were used as test organisms. The methanol extract showed more potent activity than other organic extracts, spent medium of the culture exhibited little activity against E. coli only. No inhibitory effect was found against Klebsiella pneumoniae.The broth microdilution assay gave minimum inhibitory concentrations (MIC) values ranging from 1 to 512 μg/ml. The MIC of methanol extract against S. aureus and E. coli were 128 μg/ml and 256 μg/ml, respectively.     

A newly isolated Streptomyces sp. CS392 producing three antimicrobial compounds

With the aim of isolating new microbes capable of producing strong antimicrobial substances, strain CS392 was screened from 700 soil isolates preserved in our laboratory. The strain was related to genus Streptomyces based on various characteristics. Three highly active antimicrobial compounds, C1, C2 and C3, produced by the strain were purified by solvent extraction followed by silica gel column chromatography. These compounds were highly active against various Gram-positive resistant bacteria such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Staphylococcus aureus (VRSA), and vancomycin-resistant Enterococcus (VRE). Among three, C3 was the most active against MRSA and VRSA with minimal inhibitory concentration (MIC) of 2 μg/ml while C2 and C3 had MIC values of 4 μg/ml for the strains. In case of Bacillus subtilis ATCC6633, C1 and C3 were more effective with MIC values of 0.5 μg/ml than C2 with MIC of 2 μg/ml. Those antibiotics were variably active (MIC of 4-32 μg/ml) against Micrococcus luteus ATCC 9341, Enterococcus faecalis ATCC 29212, Mycobacterium smegmatis ATCC 9341 and VRE.     

Antibacterial activity of crude methanolic extract and its fractions of aerial parts of Anthemis tinctoria

The antibacterial activity of the methanolic extract and its fractions of aerial parts of Aniheinis tinctoria (Asteraceae) was investigated against representative gram-positive Staphylococcus aureus (ATCC 25923) and Enterococcus faecalis (ATCC 29212) and gram-negative strains Escherichia coli (ATCC 25922) and Pseudomonas aeruginosa (ATCC 27853). The activity was concentrated mainly in the dichloromethane (DCM) and hexane fractions of crude methanolic extract. The 5 mg of DCM extract per disk produced 15-16 mm of inhibition zone against S. aureus and P. aeruginosa, however, no activity was found against E. faecalis and E. coli. The hexane fraction showed activity against S. aureus, P. aeruginosa and E. faecalis. As DCM fraction showed the highest antibacterial activity in the disk diffusion assay, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of only this fraction was determined against S. aureus and P. aeruginosa. These values were found to be in the range of 1.25 to 10 mg/ml.     

Antibacterial activity of extracts and neolignans from Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck

The evaluation of the activity of the aqueous and ethyl acetate extracts of the leaves of Piper regnellii was tested against gram-positive and gram-negative bacteria. The aqueous extract displayed a weak activity against Staphylococcus aureus and Bacillus subtilis with minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of 1000 micrograms/ml. The ethyl acetate extract presented a good activity against S. aureus and B. subtilis with MIC and MBC at 15.62 micrograms/ml. In contrast to the relative low MICs for gram-positive bacteria, gram-negative bacteria were not inhibited by the extracts at concentrations < or = 1000 mg/ml. The ethyl acetate extract was fractionated on silica gel into nine fractions. The hexane and chloroform fractions were active against S. aureus (MIC at 3.9 micrograms/ml) and B. subtilis (MIC at 3.9 and 7.8 micrograms/ml, respectively). Using bioactivity-directed fractionation, the hexane fraction was rechromatographed to yield the antimicrobial compounds 1, 2, 5, and 6 identified as eupomatenoid-6, eupomatenoid-5, eupomatenoid-3, and conocarpan, respectively. The pure compounds 1 and 2 showed a good activity against S. aureus with MIC of 1.56 micrograms/ml and 3.12 micrograms/ml, respectively. Both compounds presented MIC of 3.12 micrograms/ml against B. subtilis. The pure compound 6 named as conocarpan was quite active against S. aureus and B. subtilis with MIC of 6.25 micrograms/ml. The antibacterial properties of P. regnellii justify its use in traditional medicine for the treatment of wounds, contaminated through bacteria infections.     

Tobramycin (Nebramycin Factor 6): In Vitro Activity Against Pseudomonas aeruginosa

Tobramycin (factor 6 of the nebramycin complex) is a new aminoglycoside antibiotic isolated from Streptomyces tenebrarius which is active against S. aureus, Enterobacteriaceae, and Pseudomonas aeruginosa. Susceptibility to tobramycin of 96 strains of P. aeruginosa, including 45 recent isolates from blood, was studied by using agar and broth dilution methods. The minimum inhibitory concentration (MIC) for 83 of 96 strains was 3.12 mug/ml or lower in Mueller Hinton agar; MIC values were two to eight times lower in Mueller Hinton broth tests. Agar dilution MIC values were generally lower than those obtained in parallel tests with gentamicin. Killing curves obtained from serial sampling of broth cultures showed a 100- to 10,000-fold decline in viability of log-phase organisms within 30 min of exposure to the drug. Two-dimensional agar dilution tests with carbenicillin and tobramycin with 79 strains showed additive or synergistic effects; no antagonism was documented. Seventy-eight of 79 strains were inhibited by a combination of 50 mug of carbenicillin per ml and 1.56 mug of tobramycin per ml, blood levels which seem attainable in man. Tobramycin appears to be a potent, rapidly bactericidal antibiotic against P. aeruginosa and merits clinical evaluation.     

Complete genome sequence of highly multidrug-resistant Pseudomonas aeruginosa NCGM2.S1, a representative strain of a cluster endemic to Japan

We report the completely annotated genome sequence of Pseudomonas aeruginosa NCGM2.S1, a representative strain of a cluster endemic to Japan with a high level of resistance to carbapenem (MIC ≥ 128 μg/ml), amikacin (MIC ≥ 128 μg/ml), and fluoroquinolone (MIC ≥ 128 μg/ml).     

Synthesis and biological evaluation of new N-(2-hydroxy-4(or 5)-nitro/aminophenyl)benzamides and phenylacetamides as antimicrobial agents

A new series of N-(2-hydroxy-4(or 5)-nitro/aminophenyl)benzamide and phenylacetamide derivatives (1a-1n, 2a-2n) were synthesized and evaluated for antibacterial and antifungal activities against Staphylococcus aureus, Bacillus subtilis, Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli, Candida albicans, and their drug-resistant isolate. Microbiological results indicated that the compounds possessed a broad spectrum of activity against the tested microorganisms at MIC values between 500 and 1.95 microg/ml. Benzamide derivative 1d exhibited the greatest activity with MIC values of 1.95, 3.9, and 7.8 microg/ml against drug-resistant B. subtilis, B. subtilis, and S. aureus, respectively.     

Antibacterial xanthones from Kielmeyera variabilis mart. (Clusiaceae)

The bioassay-guided fractionation of stems from Kielmeyera variabilis, traditionally used in Brazilian folk medicine, yielded assiguxanthone-B (1), kielcorin (4), 2,5-dihydroxybenzoic acid (3), and a mixture of xanthones containing assiguxanthone-B (1) and 1,3,5,6-tetrahydroxy-2-prenylxanthone (2) (1:1 w/w). The xanthone mixture inhibited Staphylococcus aureus and Bacillus subtilis at a concentration of 6.25 g/ml. When tested alone, the minimal inhibitory concentration of assiguxanthone-B was 25 g/ml against B. subtilis. Kielcorin and 2,5-dihydroxybenzoic acid were inactive against both strains. None of the fractions was active against Escherichia coli or Pseudomonas aeruginosa. Viable cells of S. aureus were reduced by a 1-3 log CFU/ml within 12 h after exposure of one to eight times the MIC of the xanthone mixture. It is not known whether the tetrahydroxy-2-prenylxanthone or other components of the xanthone mixture are responsible for the main antibacterial activity or whether additive or synergistic action is involved     

Antimicrobial activities of Conyzolide and Conyzoflavone from Conyza canadensis

Antibacterial and antifungal activities of the two isolated compounds from Conyza canadensis have been reported in the current study. The two isolated compounds i.e. Conyzolide (1) and Conyzoflavone (2) were tested against six bacterial and five fungal strains, employing hole diffusion and macrodilution methods. Both the compounds showed significant activities against the tested pathogens with special reference to E. coli, P. aeruginosa, S. aureus, Trichophytom longifusus, C. albicans, and C. glaberata. Conyzolide revealed comparatively better antibacterial activity against E. coli (minimum inhibitory concentration (MIC): 25 μg/mL) in comparison to Conyzoflavone. However, in case of antifungal activities, Conyzoflavone exhibited superior antifungal activity against C. albicans (MIC: 10 μg/mL) as compared to Conyzolide.     

Screening of some plants from Northern Argentina for their antimicrobial activity

AIMS: Screening of antimicrobial activity in 25 plant species from Northern Argentina. METHODS AND RESULTS: Inhibition of microbial growth was measured by a microplate assay with an oxidation-reduction indicator (Alamar Blue). Test organisms were: Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus and Enterococcus faecium. Weak inhibitory activities (MIC=0.5 mg dry matter ml(-1)) were found in methanolic extracts of Rivina humilis, Crateva tapia, Funastrum claucum and Schinopsis balansae. Stronger bacteriostatic power was detected in Vassobia breviflora (MIC=0.25 mg ml(-1) against Staphylococcus aureus, and 0.5 mg ml(-1) against Enterococcus faecium). This activity was purified five-fold by extraction with dichloromethane, and it was found equally effective against susceptible or antibiotic-resistant strains of Staph. aureus. In addition, the purified extract was synergistic with gentamicin, and it was bactericidal at 24 h, with a concentration of 0.25 mg ml(-1). CONCLUSION: There is a significant antimicrobial activity in Vassobia breviflora. SIGNIFICANCE AND IMPACT OF THE STUDY: Further studies will be required to disclose the potential importance of these findings.     

Highly effective, water-soluble, hemocompatible 1,3-propylene oxide-based antimicrobials: poly[(3,3-quaternary/PEG)-copolyoxetanes

This study focuses on the solution antimicrobial effectiveness of a novel class of copolyoxetanes with quaternary ammonium and PEG-like side chains. A precursor P[(BBOx-m)(ME2Ox)] copolyoxetane was prepared by cationic ring-opening copolymerization of 3-((4-bromobutoxy)methyl)-3-methyloxetane (BBOx) and 3-((2-(2-methoxyethoxy)ethoxy)methyl)-3-methyloxetane (ME2Ox) to give random copolymers with 14-100 (m) mol % BBOx. Reaction of P[(BBOx-m)(ME2Ox)] with dodecyl dimethylamine gave the corresponding quaternary P[(C12-m)(ME2Ox)] polycation salts, designated C12-m, as viscous liquids in 100% yield. BBOx/ME2Ox and C12/ME2Ox ratios were obtained by (1)H NMR spectroscopy. C12-m molecular weights (M(n), 3.5-21.9 kDa) were obtained from (1)H NMR end group analysis. DSC studies up to 150 °C showed only thermal transitions between -69 and -34 °C assigned to T(g) values. Antibacterial activity for the C12-m copolyoxetanes was tested by determining minimum inhibitory concentrations (MICs) against Gram(+) Staphylococcus aureus and Gram(-) Escherichia coli and Pseudomonas aeruginosa . MIC decreased with increasing C12 mol percent, reaching a minimum in the range C12-43 to C12-60. Overall, the antimicrobial with consistently low MICs for the three tested pathogenic bacteria was C12-43: (bacteria, MIC, μg/mL) E. coli (6), S. aureus (5), and P. aeruginosa (33). For C12-43, minimum biocidal concentration (MBC) to reach 99.99% kill in 24 h required 1.5× MIC for S. aureus and 2× MIC for E. coli and P. aeruginosa . At 5× MIC against a challenge of 10(8) cfu/mL, C12-43 kills ≥99% S. aureus , E. coli , and P. aeruginosa within 1 h. C12-m copolyoxetane cytotoxicity toward human red blood cells was low, indicating good prospects for biocompatibility. The tunability of C12-m copolyoxetane compositions, effective antimicrobial behavior against Gram(+) and Gram(-) bacteria, and promising biocompatibility offer opportunities for further modification and potential applications as therapeutic agents.     

Antibacterial activity of ethanolic and aqueous extracts of Acacia aroma Gill. ex Hook et Arn

The purpose of the present study was to investigate the antibacterial activity of seven ethanolic extracts and three aqueous extracts from various parts (leaves, stems and flowers) of A. aroma against 163 strains of antibiotic multi-resistant bacteria. The disc diffusion assay was performed to evaluate antibacterial activity of the A. aroma crude extracts, against several Gram-positive bacteria (E. faecalis, S. aureus, coagulase-negative stahylococci, S. pyogenes, S. agalactiae, S. aureus ATCC 29213, E. faecalis ATCC 29212) and Gram-negative bacteria (E. coli., K. pneumoniae, P. mirabilis, E. cloacae, S. marcescens, M morganii, A. baumannii, P. aeruginosa, S. maltophilia, E. coli ATCC 35218, P. aeruginosa ATCC 27853, E. coli ATCC 25922). All ethanolic extracts showed activity against gram-positive bacteria. Among all obtained extracts, only leaf and flower fluid extracts showed activity against Gram-negative bacteria. Based on this bioassay, leaf fluid extracts tended to be the most potent, followed by flower fluid extracts. Minimal inhibitory concentration (MIC) values of extracts and antibiotics were comparatively determined by agar and broth dilution methods. Both extracts were active against S. aureus, coagulase-negative stahylococci, E. faecalis and E. faecium and all tested Gram-negative bacteria with MIC values from 0.067 to 0.308 mg/ml. In this study the minimal bactericidal concentration (MBC) values were identical or twice as high than the corresponding MIC for leaf extracts and four or eight times higher than MIC values for flower extracts. This may indicate a bactericidal effect. Stored extracts have similar antibacterial activity as recently obtained extracts. The A. aroma extracts of leaves and flowers may be useful as antibacterial agents against Gram- negative and Gram-positive antibiotic multi-resistant microorganisms.     

Antibacterial activity, structure and CMC relationships of alkanediyl alpha,omega-bis(dimethylammonium bromide) surfactants

Some alpha,omega-alkanediyl bis-dimethylammonium bromide compounds (gemini surfactants) referred as "m-s-m" have been synthesized, purified and characterized by usual spectroscopic methods. These compounds have been screened for antibacterial activity against Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. Their activity was compared. The compounds tested showed excellent in vitro antibacterial activity against Staphylococcus aureus ranging from 1.5 to 20 microg/ml and had variable activity against E. coli with minimum minimum inhibitory concentration (MIC) of 50 microg/ml. These compounds are less active against P. aeruginosa. On the other hand, contrary to the antibacterial activity of these products against S. aureus, a relation between the MIC and the critical micellar concentration (CMC) was found and relationship between chain's Length and antibacterial activity was found.     

An assessment of two Carpobrotus species extracts as potential antimicrobial agents.

For centuries, indigenous people in South Africa have used a variety of medicinal herbs to treat chronic infections. This investigation focused on two Carpobrotus species belonging to the family, Aizoaceae, in an attempt to assess their antimicrobial potential. Extracts of varying polarities of the plants were prepared and tested against Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans and Mycobacterium smegmatis. For the disc diffusion method, Ciprofloxacin (40 microg/disc) served as positive control for S. aureus, P. aeruginosa and M. smegmatis, whereas amphotericin B (25 microg/disc) was the control for C. albicans. A sample concentration of 10 mg/ml was used. Minimum inhibitory concentrations (MIC) were determined by two-fold serial dilution. Phytochemical analysis was completed to test for the presence of flavonoids, hydrolysable tannins, phytosterols and aromatic acids. The ethyl acetate extracts (21 microl of 95 mg/ml) were used for bio-autography, together with TLC analyses. Carpobrotus muirii and Carpobrotus quadrifidus showed antimicrobial activity against S. aureus and M. smegmatis in the disc diffusion method and inhibition against S. aureus and M. smegmatis was observed by clear zones on the TLC plate. This investigation confirms that extracts of these Carpobrotus species that are used as indigenous medicines, exhibit anti-bacterial activity. This scientific information can serve as an important platform for the development of inexpensive, safe and effective natural anti-infective medicines.     

Synthesis, molecular modeling and preliminary biological evaluation of a set of 3-acetyl-2,5-disubstituted-2,3-dihydro-1,3,4-oxadiazole as potential antibacterial, anti-Trypanosoma cruzi and antifungal agents

A series of 3-acetyl-2,5-disubstituted-2,3-dihydro-1,3,4-oxadiazole derivatives was synthesized and their activity screened in vitro against Staphylococcus aureus, Trypanosoma cruzi, and Candida albicans. The bioactivity was expressed as minimum inhibitory concentration (MIC) for S. aureus strains, and as fifty-percent inhibitory concentration (IC(50)) of parasite population growth for T. cruzi. A molecular modeling approach was performed to establish qualitative relationships regarding the biological data and the compounds' physicochemical properties. The 5-(4-OC(4)H(9)Ph, 5l), and 5-(4-CO(2)CH(3)Ph, 5o) derivatives were the most active compounds for S. aureus ATCC 25923 (MIC=1.95-1.25 μg/mL) and T. cruzi (IC(50)=7.91 μM), respectively. Also, a preliminary evaluation against C. albicans involving some compounds was performed and the 5-(4-CH(3)Ph, 5e) derivative was the most active compound (MIC=3.28-2.95 μg/mL). In this preliminary study, all synthesized 3-acetyl-2,5-disubstituted-2,3-dihydro-1,3,4-oxadiazole derivatives were active against all microorganisms tested.     

Comparative activity of tobramycin and gentamicin against Pseudomonas,Proteus and Providencia species.

Tobramycin is a new antibiotic resembling gentamicin. We measured the minimal inhibitory concentrations of these two antibiotics against five bacterial species that cause hospital-acquired infections and are resistant to many presently available antibiotics. The organisms tested were 500 strains of Pseudomones aeruginosa, 100 strains of each of Proteus rettgeri and Pr. morganii, 50 strains of Pr. vulgaris and 250 strains of Providencia stuartii. Tobramycin was 2 to 4 times more active than gentamicin against Ps. aeruginosa; all except 6 of 70 strains resistant to 4 mug/ml of gentamicin were sensitive to 4 mug/ml of tobramycin. The two antibiotics showed a similar degree of activity against the other four species. Tobramycin promises to be of particular value in the treatment of Pseudomonas infections.     

Proteome analyses of cellular proteins in methicillin-resistant Staphylococcus aureus treated with rhodomyrtone, a novel antibiotic candidate

The ethanolic extract from Rhodomyrtus tomentosa leaf exhibited good antibacterial activities against both methicillin-resistant Staphylococcus aureus (MRSA) and S. aureus ATCC 29213. Its minimal inhibitory concentration (MIC) values ranged from 31.25-62.5 μg/ml, and the minimal bactericidal concentration (MBC) was 250 μg/ml. Rhodomyrtone, an acylphloroglucinol derivative, was 62.5-125 times more potent at inhibiting the bacteria than the ethanolic extract, the MIC and MBC values were 0.5 μg/ml and 2 μg/ml, respectively. To provide insights into antibacterial mechanisms involved, the effects of rhodomyrtone on cellular protein expression of MRSA have been investigated using proteomic approaches. Proteome analyses revealed that rhodomyrtone at subinhibitory concentration (0.174 μg/ml) affected the expression of several major functional classes of whole cell proteins in MRSA. The identified proteins involve in cell wall biosynthesis and cell division, protein degradation, stress response and oxidative stress, cell surface antigen and virulence factor, and various metabolic pathways such as amino acid, carbohydrate, energy, lipid, and nucleotide metabolism. Transmission electron micrographs confirmed the effects of rhodomyrtone on morphological and ultrastructural alterations in the treated bacterial cells. Biological processes in cell wall biosynthesis and cell division were interrupted. Prominent changes including alterations in cell wall, abnormal septum formation, cellular disintegration, and cell lysis were observed. Unusual size and shape of staphylococcal cells were obviously noted in the treated MRSA. These pioneer findings on proteomic profiling and phenotypic features of rhodomyrtone-treated MRSA may resolve its antimicrobial mechanisms which could lead to the development of a new effective regimen for the treatment of MRSA infections.     

Influences of %T>MIC achievement probability due to the difference of the MIC measurement concentration range-analysis of meropenem for Pseudomonas aeruginosa-

We attempted to analyze any influences to %T>MIC achievement probability due to the difference of the MIC measurement concentration range of MEPM for 613 strains of Pseudomonas aeruginosa by the Monte Carlo simulation method. As for the analysis, we calculated the achievement probability of 30% and 50% for MEPM %T>MIC by the administration volume of MEPM: 250 mg, 500 mg, and 1,000 mg, the administration time: 0.5 h, and 3 h, the administration frequency: 2 times, and 3 times, and the renal excretion capability: Normal, Slight, Moderate, and High abnormal with the 3 types of MIC concentration measurement level 1) <=0.06~>=256 μg/ml: 13 levels, 2) <=0.5~>=32 μg/ml: 7 levels, and 3) <=1~>=16 μg/ml: 5 levels. As the result, we found the following findings; 1. In terms of the administration of normal renal excretion capability, 250 mg, in comparison with 500 mg and 1,000 mg, indicated the differential due to the difference of MIC measurement concentration range. 2. The administration volume of MEPM 500 mg which has been recommended shown the less differential of the achievement probability due to the difference of MIC measurement concentration range. As the renal excretion was shifted through Normal to Slight to Moderate to High abnormal, the differential of the achievement probability due to the difference of MIC measurement concentration range was gradually decreased. With these results, PK/PD analysis is possible for the 5 levels measurement concentration. It is significant that the facility using the automated microbiology analyzer can provide not only the MIC report, but also the information on the appropriate administration method for antibacterial drug by PK/PD analysis.