Individual Minimum Inhibitory Concentration Research Articles

The In vitro activity of carbapenems alone and in combination with β-lactamase inhibitors against difficult-to-treat mycobacteria; Mycobacterium tuberculosis, Mycobacterium abscessus, and Mycobacterium avium complex: A systematic review.

Difficult-to-treat mycobacterial infections are increasing globally. There is an urgent need of new treatment alternatives for multidrug-resistant Mycobacterium tuberculosis (MTB), as well as nontuberculous mycobacteria such as the Mycobacterium abscessus complex (MABC) and Mycobacterium avium complex (MAC). Recently, new carbapenems and combinations of carbapenems with β-lactamase inhibitors have become available, but activity data in vitro against mycobacteria are so far scarce. Therefore, we performed a systematic review collating the minimum inhibitory concentrations (MICs) of carbapenems, with or without a β-lactamase inhibitors for MTB, MABC, and MAC. The databases PubMed and Web of Science were searched for the relevant articles in English up until September 21, 2022. Screening of studies was performed by two independent reviewers. MIC data by recommended methods with at least five individual MICs were included. Data were reported as MIC range, MIC50, modal MIC, and/or histograms when individual MICs were available. The study protocol was registered at PROSPERO (CRD42021258537). After screening, a total of 75 studies with MIC data for carbapenems with or without β-lactamase inhibitors were included in the review. For MTB, the oral carbapenem tebipenem combined with the β-lactamase inhibitor clavulanic acid resulted in the most significant reduction of MICs. For MABC, the addition of avibactam to tebipenem resulted in a 64-fold reduction of modal MIC. Data were insufficient for the analysis of MAC. Carbapenems, and in particular the novel oral compound tebipenem, in combination with clavulanic acid for MTB and avibactam for MABC may be an untapped potential for difficult-to-treat mycobacterial infections.

Analysis of the Antimicrobial and Anti-Biofilm Activity of Natural Compounds and Their Analogues against Staphylococcus aureus Isolates.

(1) Background: Staphylococcus aureus (S. aureus) is one of the most frequent causes of biofilm-associated infections. With the emergence of antibiotic-resistant, especially methicillin-resistant S. aureus (MRSA), there is an urgent need to discover novel inhibitory compounds against this clinically important pathogen. In this study, we evaluated the antimicrobial and anti-biofilm activity of 11 compounds, including phenyl propenes and phenolic aldehydes, eugenol, ferulic acid, sinapic acid, salicylaldehyde, vanillin, cinnamoyl acid, and aldehydes, against drug-resistant S. aureus isolates. (2) Methods: Thirty-two clinical S. aureus isolates were obtained from Alkhidmat Diagnostic Center and Blood Bank, Karachi, Pakistan, and screened for biofilm-forming potential, and susceptibility/resistance against ciprofloxacin, chloramphenicol, ampicillin, amikacin, cephalothin, clindamycin, streptomycin, and gentamicin using the Kirby-Bauer disk diffusion method. Subsequently, 5 representative clinical isolates were selected and used to test the antimicrobial and anti-biofilm potential of 11 compounds using both qualitative and quantitative assays, followed by qPCR analysis to examine the differences in the expression levels of biofilm-forming genes (ica-A, fnb-B, clf-A and cna) in treated (with natural compounds and their derivatives) and untreated isolates. (3) Results: All isolates were found to be multi-drug resistant and dominant biofilm formers. The individual Minimum Inhibitory Concentration (MIC) of natural compounds and their analogues ranged from 0.75–160 mg/mL. Furthermore, the compounds, Salicylaldehyde (SALI), Vanillin (VAN), α-methyl-trans-cinnamaldehyde (A-MT), and trans-4-nitrocinnamic acid (T4N) exhibited significant (15–92%) biofilm inhibition/reduction percentage capacity at the concentration of 1–10 mg/mL. Gene expression analysis showed that salicylaldehyde, α-methyl-trans-cinnamaldehyde, and α-bromo-trans-cinnamaldehyde resulted in a significant (p < 0.05) downregulation of the expression of ica-A, clf-A, and fnb-A genes compared to the untreated resistant isolate. (4) Conclusions: The natural compounds and their analogues used in this study exhibited significant antimicrobial and anti-biofilm activity against S. aureus. Biofilms persist as the main concern in clinical settings. These compounds may serve as potential candidate drug molecules against biofilm forming S. aureus.

Antibacterial activity of Sapindus saponins against microorganisms related to food hygiene and the synergistic action mode of Sapindoside A and B against Micrococcus luteus in vitro

Sapindus saponins are natural nonionic surfactants, and they have a broad-spectrum antibacterial effect, but few studies have elucidated the antibacterial mechanism of Sapindus saponins to date. Therefore, this study screened the antibacterial activity of Sapindus saponins singly and in combination against 7 bacteria, and investigated the synergistic antibacterial action via targeting cell membrane proteins. The only combination of Sapindoside A and B (S AB ) with synergistic activity against Micrococcusluteus ( M. luteus ) was obtained, where the MICs of Sapindoside A and B were one fourth of their individual minimum inhibitory concentration (MIC). After treatment with S AB , the spectral features of protein in the cell membrane of M. luteus showed obvious changes based on Raman spectroscopy, and the constitutions of membrane proteins were changed seriously. Besides, changes in protein microenvironment was observed, and the contribution of Sapindoside A was greater than that of Sapindoside B with concentration below 2MIC. Molecular docking also demonstrated that Sapindoside A interacted with penicillin-binding protein 2, and showed higher binding energy than Sapindoside B, further indicating that the greater contribution in the synergistic action of S AB on membrane proteins. Collectively, these results showed that the synergistic antibacterial action of Sapindoside A and B against M. luteus could be achieved by attacking cell membrane proteins, and Sapindoside A played a major role, suggesting that S AB has the potential to be the natural antibacterial detergent additive in food industry, although it is limited in its scope of antimicrobial activity against foodborne bacteria. • Synergistic antibacterial combination of Sapindoside A and B (S AB ) was obtained. • Spectral features of membrane protein changed greatly based on Raman spectroscopy. • Degradation of membrane proteins and change in protein microenvironment was observed. • Contribution of Sapindoside A was greater than that of Sapindoside B in the S AB .

Synergistic Antibacterial Effect of Casein-AgNPs Combined with Tigecycline against Acinetobacter baumannii.

Acinetobacter baumannii (A. baumannii) is a common and challenging pathogen of nosocomial infections, due to its ability to survive on inanimate objects, desiccation tolerance, and resistance to disinfectants. In this study, we investigated an antibacterial strategy to combat A. baumannii via the combination of antibiotics and silver protein. This strategy used a functional platform consisting of silver nanoparticles (AgNPs) resurrected from silver-based calcium thiophosphate (SSCP) through casein and arginine. Then, the silver protein was combined with tigecycline, the first drug in glycylcycline antibiotic, to synergistically inhibit the viability of A. baumannii. The synergistic antibacterial activity was confirmed by the 96-well checkerboard method to determine their minimum inhibitory concentrations (MIC) and calculated for the combination index (CI). The MIC of the combination of silver protein and tigecycline (0.31 mg/mL, 0.16 µg/mL) was significantly lower than that of the individual MIC, and the CI was 0.59, which indicates a synergistic effect. Consequently, we integrated the detailed synergistic antibacterial properties when silver protein was combined with tigecycline. The result could make for a promising approach for the treatment of A. baumannii.

Individualised dosing algorithm and personalised treatment of high-dose rifampicin for tuberculosis.

To propose new exposure targets for Bayesian dose optimisation suited for high-dose rifampicin and to apply them using measured plasma concentrations coupled with a Bayesian forecasting algorithm allowing predictions of future doses, considering rifampicin's auto-induction, saturable pharmacokinetics and high interoccasion variability. Rifampicin exposure targets for Bayesian dose optimisation were defined based on literature data on safety and anti-mycobacterial activity in relation to rifampicin's pharmacokinetics i.e. highest plasma concentration up to 24hours and area under the plasma concentration-time curve up to 24hours (AUC0-24h ). Targets were suggested with and without considering minimum inhibitory concentration (MIC) information. Individual optimal doses were predicted for patients treated with rifampicin (10mg/kg) using the targets with Bayesian forecasting together with sparse measurements of rifampicin plasma concentrations and baseline rifampicin MIC. The suggested exposure target for Bayesian dose optimisation was a steady state AUC0-24h of 181-214h×mg/L. The observed MICs ranged from 0.016-0.125mg/L (mode: 0.064mg/L). The predicted optimal dose in patients using the suggested target ranged from 1200-3000mg (20-50mg/kg) with a mode of 1800mg (30mg/kg, n=24). The predicted optimal doses when taking MIC into account were highly dependent on the known technical variability of measured individual MIC and the dose was substantially lower compared to when using the AUC0-24h -only target. A new up-to-date exposure target for Bayesian dose optimisation suited for high-dose rifampicin was derived. Using measured plasma concentrations coupled with Bayesian forecasting allowed prediction of the future dose whilst accounting for the auto-induction, saturable pharmacokinetics and high between-occasion variability of rifampicin.

Antioxidant activity and evidence for synergism of Cannabis sativa (L.) essential oil with antimicrobial standards

The present work was conducted to study the antioxidant activity and to determine for the first time the antimicrobial potential of Moroccan Cannabis sativa essential oil (EO) singly or in combination with two conventional antibiotics (fluconazol and ciprofloxacin) against some resistant and pathogenic microorganisms. The chemical composition analyzed by means of GC/MS showed that C. sativa EO was characterized by the dominance of (E)-caryophyllene (35.0%), α-humulene (12.8%) and caryophyllene oxide (10.6%). Results from antioxidant tests showed that C. sativa EO exhibit a moderate potency (IC50 = 1.6 ± 0.1 mg/m L for 2,2- diphenyl-1-picrylhydrazyl (DPPH) assay, IC50 = 1.8 ± 0.2 mg/mL for β-carotene/linoleic acid assay, and IC50 = 0.9 ± 0.1 mg/m L for ferric reducing power assay). Regarding antimicrobial assays, the results demonstrated that C. sativa EO displayed interesting antimicrobial activities with minimum inhibitory concentration (MIC) values in the range from (1.2 to 37.8) mg/m L for tested microorganisms. The combination of the studied EO with fluconazol and ciprofloxacin showed a significant decrease in their individual MICs. In fact, at sub-inhibitory concentrations, C. sativa EO contributed to the decrease of ciprofloxacin MICs of bacterial strains by 2- to 64-fold and by16-fold regarding fluconazol MICs against Candida spp. These findings suggest that C. sativa EO can be considered as a potential source of natural antioxidant and antimicrobials, and constitute in combinational treatments a promising strategy to overcome the intense use of antibiotics against some infectious diseases.

Synergistic Activity of Essential Oils from Herbs and Spices Used on Meat Products against Food Borne Pathogens

Essential oils (EOs) could be utilized as natural agents to improve the safety of meat products. However, the high concentration required to achieve an antimicrobial effect in foods might be incompatible with their sensory acceptance. To avoid this problem, combinations of EOs provide an effective approach reducing the odds of sensory rejection. In our study, 13 EOs of herbs and spices commonly used in the seasoning of meat products were assessed for their antimicrobial activity against Salmonella spp., Listeria monocytogenes, Escherichia coli and Staphylococcus aureus. However, only 7 of them were selected to study their synergistic effect based on their antimicrobial activity and minimum inhibitory concentration (MIC) against foodborne pathogens. EOs of thyme and cinnamon presented the largest antibacterial activity against foodborne pathogens. Combinations of selected EOs displayed a synergic effect against foodborne pathogens and also an important decrease in their individual MIC. Thyme EO presented the lowest individual MIC, but its utilization in combination decreased the MIC of the other EOs. Utilization of cinnamon EO also improved the reduction of the individual MICs of the EOs of cumin and parsley. Our results suggest the potential use of EO mixtures to control foodborne pathogens in meat products. Although the individual MIC values of selected EOs decreased, the sensory impact on meat products needs to be assessed.

The redox-active drug metronidazole and thiol-depleting garlic compounds act synergistically in the protist parasite Spironucleus vortens

Spironucleus vortens is a protozoan parasite associated with significant mortalities in the freshwater angelfish, Pterophyllum scalare. Control of this parasite is especially problematic due to restrictions on the use of the drug of choice, metronidazole (MTZ), on fish farms. Use of garlic (Allium sativum) is undergoing a renaissance following experimental validations of its antimicrobial efficiency. Ajoene ((E,Z)-4,5,9-trithiadodeca-1,6,11-triene 9-oxide), is a stable transformation product of allicin, the primary biologically active component of garlic. In the current study, an ajoene oil crude extract had a minimum inhibitory concentration (MIC) of 40μg/ml against S. vortens. GC–MS and NMR spectroscopy revealed this ajoene extract contained a mixture of the (E) and (Z)-ajoene isomers along with diallyl disulphide (DADS) and diallyl trisulphide (DATS). The only component of the ajoene crude oil found to substantially inhibit S. vortens growth by optical density monitoring (Bioscreen C Reader) was (Z)-ajoene (MIC 16μg/ml). Ajoene oil acted in synergy with MTZ in vitro, reducing the individual MIC of this drug (4μg/ml) by 16-fold, and that of ajoene oil by 200-fold with a fractional inhibitory concentration (FIC) index of 0.263. This synergistic interaction was confirmed in vivo. S. vortens-infected Pterophyllum scalare angelfish dosed orally with 0.5% (v/w) MTZ combined with 0.05% (v/w) ajoene displayed a significant reduction in faecal trophozoite count, whilst those fed on 0.5% MTZ flakes (half the recommended oral dose) alone did not. This study demonstrates for the first time the synergistic interaction between the synthetic drug MTZ and natural ajoene oil both in vitro and in vivo. Future work should evaluate the potential synergy of ajoene and MTZ against MTZ-resistant bacteria and protists.

Rifaximin combined with polymyxins: A potential regimen for selective decontamination of multidrug-resistant bacteria in the digestive tract?

Selective decontamination of the digestive tract (SDD) using combinations of oral non-absorbable antibiotics has been proposed as a means of preventing multidrug-resistant (MDR) infections. The minimum inhibitory concentrations (MICs) of rifaximin (RIFAX) were determined against 262 Gram-negative and Gram-positive bacterial isolates by broth microtitre assay. Rifampicin (RIF) was used as a comparator in the analysis. Synergistic interactions between RIFAX and polymyxin B (PMB) were assessed by using the chequerboard method and calculating the fractional inhibitory concentration index (FICI). The antimicrobial activities of both RIFAX and RIF were similar with little variation in the overall MIC distributions for Gram-negative non-fermenters and Gram-positive bacteria. However, against Enterobacteriaceae higher MICs (>16mg/L) were observed for RIFAX than for RIF (50% vs 27%). Amongst the 262 isolates tested, 100 could be considered resistant to RIFAX. Overall, the combination of RIFAX and PMB was more active against all of the isolates tested compared with either drug alone, with reductions of 2-11 doubling dilutions in individual MICs. Potent synergy was observed with the RIFAX+PMB combination using FICI criteria (FICI range 0.02-0.5). The data presented here suggest that combination therapy may be significantly more effective against isolates with RIFAX and/or PMB resistance and could be considered as part of a SDD regimen aimed at reducing enteric carriage of MDR pathogens in colonised and infected patients.

Synergistic antimicrobial profiling of violacein with commercial antibiotics against pathogenic micro-organisms

Context: Chromobacterium violaceum Bergonzini (Neisseriaceae), a Gram-negative bacterium, secretes a spectacular pigment called violacein. Violacein is a quorum-sensing metabolite and is also an active antimicrobial, anticancer agent. However, its efficiency as a potential drug, alone or in synergy with other active principles, has not being completely exploited. With the advent of different multi-drug resistant strains, it becomes essential to find a new natural product(s) that could be effectively used as a therapeutic agent.Objective: This work focused on the extraction of violacein from an isolated strain of C. violaceum and determined the combinatory effect of violacein with commercial antibiotics against various pathogens.Materials and methods: Violacein production was optimized and was later extracted using ethanol and characterized by liquid chromatography-mass spectrometry and infrared spectroscopy. Then, individual minimum inhibitory concentration (MIC) values for each of the antibiotics were determined followed by violacein–commercial antibiotics (1:1) combinations, tested at different concentrations starting from 500 to 1 µg/ml against major pathogens.Results and discussion: The individual MIC data for violacein was found to be 5.7 µg/ml (Staphylococcus aureus), 15.6 µg/ml (Klebsiella pneumoniae), 18.5 µg/ml (Pseudomonas aeruginosa), 20.0 µg/ml (Vibrio cholerae) and 5.7 µg/ml (Salmonella typhi). Violacein–gentamicin and violacein-cefadroxil combinations had MIC of 1.0 µg/ml against S. aureus. Most violacein–macrolide and violacein--aminoglycoside class combinations revealed fractional inhibitory concentration indices (FICI) of <0.5, thus exhibiting synergism. Furthermore, violacein–azithromycin and violacein–kanamycin combination, exhibited significant synergy (FICI-0.3) against S. typhi.Conclusion: Violacein works synergistically with most commercial antibiotics and could be used as drug in combination with other antimicrobial agents.

In vitro effect of three-drug combinations of antituberculous agents against multidrug-resistant Mycobacterium tuberculosis isolates

Multidrug resistance has become a problem in the management of tuberculosis, leading to an urgent need for research related to new regimens including the currently available drugs. The objectives of this study were: (i) to study the effect of the following second-choice three-drug combinations against multidrug-resistant (MDR) and drug-susceptible clinical isolates (levofloxacin, linezolid and ethambutol; levofloxacin, amikacin and ethambutol; and levofloxacin, linezolid and amikacin); and (ii) to compare the effect of these combinations with an isoniazid, rifampicin and ethambutol combination against drug-susceptible clinical isolates. A total of 9 MDR clinical and 12 drug-susceptible isolates (11 clinical isolates and the H37Rv reference strain) were studied using an adaptation of the chequerboard assay. The fractional inhibitory concentration index (FICI) was calculated as follows: FICI=FICA+FICB+FICC=A/MICA+B/MICB+C/MICC, where A, B and C are the minimum inhibitory concentrations (MICs) of each antibiotic in combination and MICA, MICB and MICC are the individual MICs. The FICI was interpreted as synergism when the value was <0.75. The FICI of all the combinations ranged from 1.5 to 3, showing indifferent activity. No differences were found between MDR and drug-susceptible isolates, or between the second-choice combinations and the fourth combination against drug-susceptible isolates. In conclusion, the second-choice drugs are equally effective as the combination of isoniazid, rifampicin and ethambutol.

Synergistic effects of miconazole and polymyxin B on microbial pathogens.

The therapeutic value of antibiotics depends on the susceptibility of the infecting microorganism and the pharmacological profile of the drugs. To assess the value of an antibiotic combination of polymyxin B and miconazole this study examined the in vitro synergistic potential of the two drugs on Gram-negative and Gram-positive bacteria and yeast. Antifungal and antibacterial activity was tested by minimum inhibitory concentration (MIC) of broth macrodilution and urea broth microdilution, by fluorescence microscopy and flow cytometry. Synergism was calculated using the fractional inhibitory concentration index (FICi). With Staphylococcus intermedius as target we found up to an eightfold reduction of the individual MICs when both drugs were combined. However, the FICi was 0.63 suggesting no real interaction between the two drugs. With Escherichia coli, Pseudomonas aeruginosa, and Malassezia pachydermatis as targets the antimicrobial drug combination reduced the MICs of polymyxin B and miconazole from fourfold to hundredfold resulting in FICi between 0.06 and 0.5 which defines a synergistic action. Thus, if polymyxin B and miconazole are combined their effect is greater than the sum of the effects observed with polymyxin B and miconazole independently, revealing bactericidal and fungicidal synergism. Our results indicate a strong therapeutic value for the combination of these antimicrobial agents against Gram-negative bacteria and yeast and a weaker value against Gram positive bacteria for clinical situations where these pathogens are involved.

Modelling the inhibition of sorbic and benzoic acids on a native yeast cocktail from table olives

The single and combined effects, in a synthetic medium at selected pH values, of sorbic and benzoic acids on a yeast cocktail ( Saccharomyces cerevisiae, Pichia anomala, Issatchenkia occidentalis, and Candida diddensiae, isolated from table olives) have been studied. Applying the checkerboard method the minimum inhibitory concentration (MIC) obtained for the respective individual preservatives (expressed as undissociated acid) were: sorbic acid, 5.94, 3.85 and 3.19 mM at pH of 4.5, 4.0, and 3.5, respectively; and benzoic acid, not detected (at total 20.5 mM), 10.40 and 6.83 mM, respectively, for the same pH levels. The estimated fractional inhibitory concentrations (FIC) indexes showed additive effects between inhibitors. Fractional area (fa), modelled by the (extended) Lambert and Lambert [2003. A model for the efficacy of combined inhibitors. J. Appl. Microbiol. 95, 734–743] equation (ELPM), also showed additives of both preservatives but different shapes in the dose–response curves; the individual MIC (as undissociated acid) deduced from this method were: 5.60, 3.31, and 3.26 mM for sorbic acid at pH of 4.5, 4.0, and 3.5, respectively; and 29.65 (extrapolated), 10.00, and 6.25 mM for benzoic acid at the same pH levels. Mixtures above the curves connecting the limits (MIC) at each pH were also inhibitory. There was agreement between MIC values from FIC and ELPM, although the last one provided further information on the inhibition behaviour. I. occidentalis was the most resistant yeast of the cocktail to sorbic and benzoic acids.

Antibiotic MIC/MBC analysis of Bacillus-based commercial insecticides: use of bioreduction and DNA-based assays.

Minimum inhibitory concentration (MIC) assays, monitored by colony counts, growth (turbidity) and bioreduction of non-toxic XTT [2,3-bis (2-methoxy-4-nitro-5-sulphophenyl)-2H-tetrazolium-5-carboxanilide, inner salt], were used to assess the performance of several types of antibiotics against: (1) commercial BT products made from scale-up sporulation phase cultures of Bacillus thuringiensis subsp israelensis (Bti) and subsp kurstaki (Btk); (2) vegetative cells derived from these BT products; and (3) Gram-positive and Gram-negative bacteria used as controls. The XTT-kinetic assay improved sensitivity and early reading of MIC breakpoints. The conventional colony count method for determining minimal bactericidal concentration (MBC) was used to validate a multi-sample dot-blot assay in which organisms in individual MIC assays are trapped free of residual antibiotic and their viability is estimated by in situ conversion of MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] to insoluble formazan. Tolerance (MBC/MIC) for most antibiotics was low (</=4). Resistance to beta-lactams was attributed to beta-lactamase activity in both BT products and cultures derived from them. MIC and MBC breakpoints in spore-based assays were also approximated by changes in genome copy, using delta-endotoxin and beta-lactamase genes as probes. The DNA assays are effective for monitoring and authenticating organisms in microbe-containing biotechnology products.