Antibacterial activity and mechanism of action of trigonostemone against Staphylococcus aureus and Bacillus cereus

@#In our present study, the hexane fraction from the root of the Thai medicinal plant Strophioblachia fimbricalyx Boerl. was purified and the purification led to the isolation of 3-acetylaleuritolic acid, trigonostemone and 3,6,9-trimethoxyphenanthropolone. The aims of this work are to evaluate antibacterial activity of these three isolated compounds from our local plant and to study their mechanism of actions toward target pathogenic bacteria (Gram positive and Gram negative bacteria). The antibacterial activity of isolated compounds was primary screened by agar well diffusion method and the active compound was subjected to determine for MIC and MBC values by microdilution method. The kinetic study of the bacteriostatic or bactericidal activity time-kill experiment (24 h) and mechanism of action on cell morphology toward target bacteria detected by scanning electron microscope of the active compound were further evaluated. Results indicate that among the tested three compounds, trigonostemone was the only active one. It exhibited an inhibitory effect on the growth of Gram positive bacteria, methicillin-susceptible Staphylococcus aureus (MSSA) DMST 2933, methicillin-resistant S. aureus (MRSA) DMST 20651 and Bacillus cereus ATCC 11778 with the MIC/MBC values of 12.5/25.0, 6.25/6.25 and 6.25/6.25 mg/mL, respectively. Trigonostemone possessed time- and concentration-dependent bacteriostatic activity against S. aureus (MSSA) DMST 2933 and bactericidal activity against B. cereus ATCC 11778. It caused bacteriostatic activity against S. aureus (MSSA) DMST 2933 at the concentration of 2 × MIC by changing cell morphology and bactericidal activity against B. cereus ATCC 11778 at the concentration of 2 × MIC after 4 h by inducing cell size variations at the concentrations of 2 × MIC, respectively. This finding suggests that trigonostemone isolated from the root of S. fimbricalyx has a potential to be used as natural antibacterial compound against S. aureus (MSSA) DMST 2933 and B. cereus ATCC 11778 bacterial strains.

Evaluation of Synergistic Effect of Combined Treatment with Linalool and Colistin on Multidrug-Resistant Acinetobacter baumannii to Expand Candidate for Therapeutic Option / 대한임상미생물학회지

BACKGROUND: Acinetobacter baumannii infection is a significant health problem worldwide due to increased drug resistance. The limited antimicrobial alternatives for the treatment of severe infections by multidrug-resistant A. baumannii (MDRAB) make the search for other therapeutic options more urgent. Linalool, the major oil compound in Coriandrum sativum, was recently found to have high antibacterial activity against A. baumannii. The purpose of this study was to investigate the synergistic effect of linalool and colistin combinations against MDRAB and extensively drug-resistant A. baumannii (XDRAB).METHODS: A total of 51 strains of A. baumannii clinical isolates, consisting of 10 MDRAB and 41 XDRAB were tested. We determined the minimum inhibitory concentration (MIC) of linalool for the test strains using the broth microdilution method and searched for interactions using the time-kill assay.RESULTS: The time-kill assay showed that the linalool and colistin combination displayed a high rate of synergy (92.1%) (by synergy criteria 2), low rate of indifference (7.8%), and a high rate of bactericidal activity (74.5%) in the 51 clinical isolates of A. baumannii. The synergy rates for the linalool and colistin combination against MDRAB and XDRAB were 96% and 92.1%, respectively. No antagonism was observed for the linalool and colistin combination.CONCLUSION: The combination of linalool and colistin showed a high synergy rate, which may be beneficial for controlling MDRAB infections. Therefore, this combination is a good candidate for in vivo studies to assess its efficacy in the treatment of MDRAB infections.

Time-kill assay and post-antibiotic effect of acetone extract from the stem bark of Canarium odontophyllum against Methicillin-resistant Staphylococcus aureus Mu50 strain

Canarium odontophyllum Miq. is an indigenous fruit found in Sarawak, Malaysia. Methicillin-resistant Staphylococcusaureus (MRSA) is a deadly pathogen that causes to hospital (health-care-acquired MRSA [HA-MRSA]) and community(CA-MRSA) infections worldwide. Vancomycin has been the therapeutic drug of choice against MRSA, but unfortunatelythis pathogen has developed some degree of resistance to vancomycin. This research aimed to evaluate the antimicrobialactivity of the stem bark extract of C. odontophyllum against MRSA Mu50 strain. The minimum inhibitory concentration(MIC) and minimum bactericidal concentration (MBC) of extract and vancomycin against MRSA were determined usingbroth microdilution method and streak plate method. The rate of killing by the extract against Mu50 strain was determinedusing time-kill assay (TKA) at ×1 MIC, ×2 MIC, ×4 MIC, and ×8 MIC of the extract. The post-antibiotic effect (PAE) timeof extract ×10 MIC against MRSA was also investigated. The extract exhibited bacteriostatic effect against MRSA Mu50strain with MIC and MBC values of 1.563 mg/ml and 3.125 mg/ml, respectively. From TKA analysis, the extract was notcapable of killing the Mu50 strain at ×1 MIC and ×2 MIC, but it displayed bactericidal activity at higher concentrationstested. Interestingly, the acetone stem bark extract of C. odontophyllum at ×4 MIC showed comparable time-killingkinetic with the standard antibiotic in the study. The PAE time of the extract was 3.6 ± 0.51 h against MRSA Mu50compared to vancomycin at 2.4 ± 0.68 h. In conclusion, the stem bark acetone extract from C. odontophyllum demonstratedconcentration-dependent bactericidal effect with prolonged PAE time against MRSA Mu50 strain.

In vitro assessment of the synergism between extracts of Cocos nucifera husk and some standard antibiotics

Objective To evaluate the interactions between the crude extracts of Cocos nucifera (C. nucifera) and six front line antibiotics (ampicillin sodium salt, penicillin G sodium, amoxicillin, chloramphenicol, ciprofloxacin and tetracycline hydrochloride), against some bacterial pathogens linked with human infection. Methods The pulverized husk of C. nucifera was dissolved in 95% n-hexane and extracted using Soxhlet extraction method and sterile distilled water (aqueous). The antibacterial susceptibility of the crude extracts of C. nucifera was tested against environmental and clinical strains (6) obtained from the South African Bureau of Standards (SABS), Vibrio (6) and Listeria pathogens (6). The agar-well diffusion method was used for screening the extracts for their antibacterial activity. The minimum inhibitory concentration and minimum bactericidal concentration of the extracts were determined. Time-kill assay was used to evaluate bactericidal and/or bacteriostatic activity. The synergistic effect of the crude extracts and antibiotics was assessed and evaluated by adopting the checkerboard methods. Results With the time-kill assay, the highest bactericidal activity was observed on Vibrio fluvialis EL041 with a −5.6 ± 0.2 log

Synergistic interaction of Helichrysum pedunculatum leaf extracts with antibiotics against wound infection associated bacteria

The effect of combinations of the crude methanolic extract of the leaves of Helichrysum pedunculatum and eight first-line antibiotics were investigated by time kill assays against a panel of bacterial strains that have been implicated in wound infections. The plant extract showed appreciable antibacterial activities against the test bacteria with zones of inhibition ranging between 18 and 27 mm, and minimum inhibitory concentrations (MICs) varying between 0.1 and 5.0 mg/ml. The MICs of the test antibiotics range between 0.001 and 0.412 mg/ml, and combination of the plant extract and the antibiotics resulted in reduction of bacterial counts by between 0 and 6.63 Log10 cfu/ml. At V2 MIC, 56.81 percent synergy; 43.19 percent indifference and no antagonism were observed, and at MIC levels, 55.68 percent synergy; 44.32 percent indifference and no antagonism were observed when the extracts were combined with eight different antibiotics. In all, 60 percent of the interactions were synergistic. All combination regimes on Staphylococcus aureus ATCC 6538 yielded no synergy, neither was antagonism detected in any of the assays. We propose that extracts of the leaves of Helichrysum pedunculatum could be of relevance in combination therapy and as a source of resistance modifying principies that could be useful as treatment options for persistent wound infections.

Synergy of Arbekacin-based Combinations Against Vancomycin Hetero-intermediate Staphylococcus aureus

This study was undertaken to evaluate the in vitro activities of arbekacin-based combination regimens against vancomycin hetero-intermediate Staphylococcus aureus (hetero-VISA). Combinations of arbekacin with vancomycin, rifampin, ampicillin-sulbactam, teicoplanin, or quinipristin-dalfopristin against seven hetero-VISA strains and two methicillin-resistant S. aureus strains were evaluated by the time-kill assay. The combinations of arbekacin with vancomycin, teicoplanin, or ampicillinsulbactam showed the synergistic interaction against hetero-VISA strains. Data suggest that these arbekacin-based combination regimens may be useful candidates for treatment options of hetero-VISA infections.