Staphylococcus pseudintermedius and Pseudomonas aeruginosa Lubbock Chronic Wound Biofilm (LCWB): a suitable dual-species model for in vitro studies.

Antimicrobial treatment of methicillin-resistant Staphylococcus pseudintermedius associated with canine wounds represents an important challenge. The aim of this study was to create a canine wound infection model, Lubbock Chronic Wound Biofilm (LCWB), with a focus on S. pseudintermedius, drawing inspiration from the established human model involving Staphylococcus aureus. Methicillin-resistant S. pseudintermedius 115 (MRSP) and Pseudomonas aeruginosa 700 strains, isolated from dog wounds, were used to set up the LCWB at 24, 48 and 72 h. The LCWBs were evaluated in terms of volume, weight, and microbial CFU/mg. The microbial spatial distribution in the LCWBs was assessed by SEM and CLSM imaging. The best incubation time for the LCWB production in terms of volume (3.38 cm3 ± 0.13), weight (0.86 gr ± 0.02) and CFU/mg (up to 7.05 × 106 CFU/mg ± 2.89 × 105) was 48 h. The SEM and CLSM images showed a major viable microbial colonization at 48 h with non-mixed bacteria with a prevalence of MRSP on the surface and P. aeruginosa 700 in the depth of the wound. The obtained findings demonstrate the capability of S. pseudintermedius to grow together P. aeruginosa in the LCWB model, representing the suitable model to reproduce the animal chronic wound in vitro.

Complex magnetic fields represent an eco-sustainable technology to counteract the resistant Candida albicans growth without affecting the human gingival fibroblasts.

Novel technologies such as complex magnetic fields-CMFs represent an eco-sustainable proposal to counteract the infection associated to resistant microorganisms. The aim of this study was to evaluate the effect of two CMF programs (STRESS, ANTIBACTERIAL) against clinical antifungal resistant C. albicans also evaluating their uneffectiveness on gingival fibroblasts (hGFs). The STRESS program was more efficacious on C. albicans biofilm with up to 64.37% ± 10.80 of biomass and up to 99.19% ± 0.06 CFU/ml reductions in respect to the control also inducing an alteration of lipidic structure of the membrane. The MTT assay showed no CMFs negative effects on the viability of hGFs with a major ROS production with the ANTIBACTERIAL program at 3 and 24 h. For the wound healing assay, STRESS program showed the best effect in terms of the rate migration at 24 h, showing statistical significance of p < 0.0001. The toluidine-blue staining observations showed the typical morphology of cells and the presence of elongated and spindle-shaped with cytoplasmic extensions and lamellipodia was observed by SEM. The ANTIBACTERIAL program statistically increased the production of collagen with respect to control and STRESS program (p < 0.0001). CMFs showed a relevant anti-virulence action against C. albicans, no cytotoxicity effects and a high hGFs migration rate. The results of this study suggest that CMFs could represent a novel eco-sustainable strategy to counteract the resistant yeast biofilm infections.

In Vitro Evaluation of Candida albicans Adhesion on Heat-Cured Resin-Based Dental Composites.

Microbial adhesion on dental restorative materials may jeopardize the restorative treatment long-term outcome. The goal of this in vitro study was to assess Candida albicans capability to adhere and form a biofilm on the surface of heat-cured dental composites having different formulations but subjected to identical surface treatments and polymerization protocols. Three commercially available composites were evaluated: GrandioSO (GR), Venus Diamond (VD) and Enamel Plus HRi Biofunction (BF). Cylindrical specimens were prepared for quantitative determination of C. albicans S5 planktonic CFU count, sessile cells CFU count and biomass optical density (OD570 nm). Qualitative Concanavalin-A assays (for extracellular polymeric substances of a biofilm matrix) and Scanning Electron Microscope (SEM) analyses (for the morphology of sessile colonies) were also performed. Focusing on planktonic CFU count, a slight but not significant reduction was observed with VD as compared to GR. Regarding sessile cells CFU count and biomass OD570 nm, a significant increase was observed for VD compared to GR and BF. Concanavalin-A assays and SEM analyses confirmed the quantitative results. Different formulations of commercially available resin composites may differently interact with C. albicans. The present results showed a relatively more pronounced antiadhesive effect for BF and GR, with a reduction in sessile cells CFU count and biomass quantification.

Helicobacter pylori Dormant States Are Affected by Vitamin C.

Helicobacter pylori colonizes human gastric mucosa, overcoming stressful conditions and entering in a dormant state. This study evaluated: (i) H. pylori's physiological changes from active to viable-but-non-culturable (VBNC) and persister (AP) states, establishing times/conditions; (ii) the ability of vitamin C to interfere with dormancy generation/resuscitation. A dormant state was induced in clinical MDR H. pylori 10A/13 by: nutrient starvation (for VBNC generation), incubating in an unenriched medium (Brucella broth) or saline solution (SS), and (for AP generation) treatment with 10xMIC amoxicillin (AMX). The samples were monitored after 24, 48, and 72 h, 8-14 days by OD600, CFUs/mL, Live/Dead staining, and an MTT viability test. Afterwards, vitamin C was added to the H. pylori suspension before/after the generation of dormant states, and monitoring took place at 24, 48, and 72 h. The VBNC state was generated after 8 days in SS, and the AP state in AMX for 48 h. Vitamin C reduced its entry into a VBNC state. In AP cells, Vitamin C delayed entry, decreasing viable coccal cells and increasing bacillary/U-shaped bacteria. Vitamin C increased resuscitation (60%) in the VBNC state and reduced the aggregates of the AP state. Vitamin C reduced the incidence of dormant states, promoting the resuscitation rate. Pretreatment with Vitamin C could favor the selection of microbial vegetative forms that are more susceptible to H. pylori therapeutical schemes.

Biofilms in Chronic Wound Infections: Innovative Antimicrobial Approaches Using the In Vitro Lubbock Chronic Wound Biofilm Model.

Chronic wounds have harmful effects on both patients and healthcare systems. Wound chronicity is attributed to an impaired healing process due to several host and local factors that affect healing pathways. The resulting ulcers contain a wide variety of microorganisms that are mostly resistant to antimicrobials and possess the ability to form mono/poly-microbial biofilms. The search for new, effective and safe compounds to handle chronic wounds has come a long way throughout the history of medicine, which has included several studies and trials of conventional treatments. Treatments focus on fighting the microbial colonization that develops in the wound by multidrug resistant pathogens. The development of molecular medicine, especially in antibacterial agents, needs an in vitro model similar to the in vivo chronic wound environment to evaluate the efficacy of antimicrobial agents. The Lubbock chronic wound biofilm (LCWB) model is an in vitro model developed to mimic the pathogen colonization and the biofilm formation of a real chronic wound, and it is suitable to screen the antibacterial activity of innovative compounds. In this review, we focused on the characteristics of chronic wound biofilms and the contribution of the LCWB model both to the study of wound poly-microbial biofilms and as a model for novel treatment strategies.

Spread of Multidrug-Resistant Microorganisms.

Multidrug-resistant organisms (MDRO) are bacteria that exhibit acquired resistance to multiple antibiotics, reducing the efficacy of antimicrobial therapies [...].

Antimicrobial Combined Action of Graphene Oxide and Light Emitting Diodes for Chronic Wound Management.

Innovative non-antibiotic compounds such as graphene oxide (GO) and light-emitting diodes (LEDs) may represent a valid strategy for managing chronic wound infections related to resistant pathogens. This study aimed to evaluate 630 nm LED and 880 nm LED ability to enhance the GO antimicrobial activity against Staphylococcus aureus- and Pseudomonas aeruginosa-resistant strains in a dual-species biofilm in the Lubbock chronic wound biofilm (LCWB) model. The effect of a 630 nm LED, alone or plus 5-aminolevulinic acid (ALAD)-mediated photodynamic therapy (PDT) (ALAD-PDT), or an 880 nm LED on the GO (50 mg/l) action was evaluated by determining the CFU/mg reductions, live/dead analysis, scanning electron microscope observation, and reactive oxygen species assay. Among the LCWBs, the best effect was obtained with GO irradiated with ALAD-PDT, with percentages of CFU/mg reduction up to 78.96% ± 0.21 and 95.17% ± 2.56 for S. aureus and P. aeruginosa, respectively. The microscope images showed a reduction in the cell number and viability when treated with GO + ALAD-PDT. In addition, increased ROS production was detected. No differences were recorded when GO was irradiated with an 880 nm LED versus GO alone. The obtained results suggest that treatment with GO irradiated with ALAD-PDT represents a valid, sustainable strategy to counteract the polymicrobial colonization of chronic wounds.

Complex Chronic Wound Biofilms Are Inhibited in vitro by the Natural Extract of Capparis spinose.

Resistant wound microorganisms are becoming an extremely serious challenge in the process of treating infected chronic wounds, leading to impaired healing. Thus, additional approaches should be taken into consideration to improve the healing process. The use of natural extracts can represent a valid alternative to treat/control the microbial infections in wounds. This study investigates the antimicrobial/antivirulence effects of Capparis spinose aqueous extract against the main chronic wound pathogens: Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans. The extract shows phenolic characterization with rutin (1.8 ± 0.14 µg/mg) as the major compound and antibacterial effect against bacteria (S. aureus PECHA 10 MIC 6.25%; P. aeruginosa PECHA 4 MIC 12.50%) without action against C. albicans (MIC and MFC ≥ 50%). Capparis spinose also shows a significant antivirulence effect in terms of antimotility/antibiofilm actions. In particular, the extract acts (i) on P. aeruginosa both increasing its swimming and swarming motility favoring the planktonic phenotype and reducing its adhesive capability, (ii) on S. aureus and P. aeruginosa biofilm formation reducing both the biomass and CFU/ml. Furthermore, the extract significantly displays the reduction of a dual-species S. aureus and P. aeruginosa Lubbock chronic wound biofilm, a complex model that mimics the realistic in vivo microbial spatial distribution in wounds. The results suggest that C. spinose aqueous extract could represent an innovative eco-friendly strategy to prevent/control the wound microbial infection.

Antimicrobial Peptide L18R Displays a Modulating Action against Inter-Kingdom Biofilms in the Lubbock Chronic Wound Biofilm Model.

Chronic wound infections represent an important health problem due to the reduced response to antimicrobial treatment of the pathogens organized in structured biofilms. This study investigated the effects of the previously described antifungal peptide L18R against three representative wound pathogens: Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans. The antimicrobial activity of L18R was evaluated (i) against single planktonic microbial populations; (ii) on single, dual, and triadic species of biofilms in both the early stage and mature stage; and (iii) in the polymicrobial Lubbock chronic wound biofilm (LCWB) model, mimicking spatial microbial colonization. This study used the evaluation of CFUs, biofilm biomass detection, and confocal and scanning electron microscopy analysis. L18R showed a significant antimicrobial activity against planktonic microorganisms and was able to differentially reduce the biomass of monomicrobial biofilms. No reduction of biomass was observed against the polymicrobial biofilm. In mature LCWB, L18R caused a moderate reduction in total CFU number, with a variable effect on the different microorganisms. Microscopy images confirmed a predominant presence of P.aeruginosa and a lower percentage of C. albicans cells. These findings suggest a modulating action of L18R and recommend further studies on its potential role in chronic wound management in association with conventional antibiotics or alternative treatments.

Searching for New Tools to Counteract the Helicobacter pylori Resistance: The Positive Action of Resveratrol Derivatives.

The drug-resistance phenomenon in Helicobacter pylori underlines the need of novel strategies to improve the eradication rate including alternative treatments combining antibiotic and non-antibiotic compounds with synergistic action. In this study, the antibacterial (MIC/MBC) and anti-virulence effects (biofilm reduction and swarming motility inhibition) of resveratrol-RSV and new synthetized RSV-phenol derivatives, with a higher bioavailability, alone and combined with levofloxacin-LVX were evaluated against resistant H. pylori clinical strains. The experiments were confirmed in vivo using the Galleria mellonella model. Among the studied RSV derivatives, RSV-3 and RSV-4 possessed higher antibacterial activity with respect to RSV (MICs from 6.25 to 200 µg/mL and from 3.12 to 200 µg/mL, respectively). RSV, RSV-3, and RSV-4 were able to synergize with LVX restoring its effect in two out of seven clinical resistant strains tested for the study. RSV, RSV-3, and RSV-4, alone and with LVX at sub-MIC and sub-synergistic concentrations, significantly reduced the biofilm formation. Moreover, RSV-3 and RSV-4 reduced the H. pylori swarming motility on soft agar. RSV, RSV-3, and RSV-4 were non-toxic for G. mellonella larvae and displayed a protective effect against H. pylori infection. Overall, RSV-phenol derivatives should be considered interesting candidates for innovative therapeutic schemes to tackle the H. pylori antibiotic resistance.

Prebiotic Combinations Effects on the Colonization of Staphylococcal Skin Strains.

BACKGROUND: An unbalanced skin microbiota due to an increase in pathogenic vs. commensal bacteria can be efficiently tackled by using prebiotics. The aim of this work was to identify novel prebiotic combinations by exerting species-specific action between S. aureus and S. epidermidis strains. METHODS: First, the antimicrobial/antibiofilm effect of Xylitol-XYL and Galacto-OligoSaccharides-GOS combined with each other at different concentrations (1, 2.5, 5%) against S. aureus and S. epidermidis clinical strains was evaluated in time. Second, the most species-specific concentration was used to combine XYL with Fructo-OligoSaccharides-FOS, IsoMalto-Oligosaccharides-IMO, ArabinoGaLactan-LAG, inulin, dextran. Experiments were performed by OD600 detection, biomass quantification and LIVE/DEAD staining. RESULTS: 1% XYL + 1% GOS showed the best species-specific action with an immediate antibacterial/antibiofilm action against S. aureus strains (up to 34.54% ± 5.35/64.68% ± 4.77) without a relevant effect on S. epidermidis. Among the other prebiotic formulations, 1% XYL plus 1% FOS (up to 49.17% ± 21.46/37.59% ± 6.34) or 1% IMO (up to 41.28% ± 4.88/36.70% ± 10.03) or 1% LAG (up to 38.21% ± 5.31/83.06% ± 5.11) showed antimicrobial/antibiofilm effects similar to 1% XYL+1% GOS. For all tested formulations, a prevalent bacteriostatic effect in the planktonic phase and a general reduction of S. aureus biofilm formation without loss of viability were recorded. CONCLUSION: The combinations of 1% XYL with 1% GOS or 1% FOS or 1% IMO or 1% LAG may help to control the balance of skin microbiota, representing good candidates for topic formulations.

Graphene Oxide affects Staphylococcus aureus and Pseudomonas aeruginosa dual species biofilm in Lubbock Chronic Wound Biofilm model.

Chronic wound management becomes a complex procedure because of the persistence of forming biofilm pathogens that do not respond to antimicrobial treatment. The aim of this paper is to detect the Graphene Oxide-GO effect on Staphylococcus aureus and Pseudomonas aeruginosa dual species wound biofilm in Lubbock Chronic Wound Biofilm-LCWB model. LCWB is a recognized model that mimics the spatial microbial colonization into chronic wounds and reproduces the wound and its clot. Staphylococcus aureus PECHA 10 and P. aeruginosa PECHA 4, are the pathogens used in the study. The GO effect on both in forming and mature biofilms, is detected by the evaluation of the CFU/mg reduction, the cell viability and ultrastructural analysis of the treated LCWBs. Graphene Oxide, at 50 mg/l, shows a significant antibiofilm effect in forming and mature LCWBs. In particular, during the biofilm formation, GO reduces the S. aureus and P. aeruginosa growth of 55.05% ± 4.73 and 44.18% ± 3.91 compared to the control. In mature biofilm, GO affects S. aureus and P. aeruginosa by reducing their growth of 70.24% ± 4.47 and 63.68% ± 17.56, respectively. Images taken by SEM show that GO display a disaggregated microbial effect also disrupting the fibrin network of the wound-like biofilm framework. In conclusion, GO used against microorganisms grown in LCWB, displays a significant inhibitory action resulting in a promising tool for potential application in wound management.

Near-infrared NIR irradiation and sodium hypochlorite: An efficacious association to counteract the Enterococcus faecalis biofilm in endodontic infections.

New strategies are necessary for the prevention of endodontic infections caused by Enterococcus faecalis, a common resistant pathogen and biofilm producer. Aim of the present study was to compare the effects of Near-Infrared (NIR) Light-Emitting Diode (LED) irradiation and different concentrations of sodium hypochlorite (NaOCl) alone or combined to each other on the E. faecalis biofilm, on artificial and human dentin surfaces. E. faecalis ATCC 29212 preformed biofilms, on polystyrene wells and on dentin discs, were treated with 880 nm NIR irradiation and NaOCl at 4%, 2.5%, 1% and 0.5% alone and combined to each other (NIR irradiation plus NaOCl 1% or 0.5%) at 5 and 10 min. Treated biofilms were compared to the controls for (i) biofilm biomass evaluation, (ii) CFU count for the quantification of cultivable cells and (iii) cells viability. All the detected experimental conditions displayed a significant reduction of biofilm biomass (p < 0.001) and CFUs/mL (p < 0.01) in respect to the controls on both tested surfaces. The effects on the E. faecalis biomass, colony count and cell viability were not time-dependent except for NaOCl 2.5% and 1% in the biofilm biomass reductions on human dentin discs. NIR-LED irradiation alone showed a reduction of E. faecalis aggregates without interfering with cell viability whereas NaOCl alone expressed a killing effect in a concentration dependent way. The combination of NIR-LED irradiation with NaOCl 1% and 0.5% displayed a double effect of cluster disaggregation and cell killing. In particular, NIR-LED irradiation combined with NaOCl 0.5% displayed an anti-biofilm activity major than those expressed by NaOCl 0.5% alone (p = 0.001) with a reduction of biomass 93% vs 71% and 97% vs 25% after 10 min, on polystyrene wells and human dentin discs, respectively. The innovative use of NIR-LED irradiation combined at short times with low concentration of NaOCl (1% and 0.5%) is capable to reach a significant effect on E. faecalis biofilm, especially on human dentin discs.

Porphyromonas Gingivalis Load is Balanced by 0.20% Chlorhexidine Gel. A Randomized, Double-Blind, Controlled, Microbiological and Immunohistochemical Human Study.

Microbial contamination could compromise the stability of dental implants increasing the risk of inflammatory reactions in the surrounding soft tissues. In this human, randomized, double-blind, clinical study, the presence of Porphyromonas gingivalis on the healing abutment and the inflammatory infiltrate surrounding peri-implant soft tissues were investigated. Experiments were done in order to clarify the effect of 0.20% chlorhexidine (CHX) versus placebo, applied during each rehabilitation stage. Thirty patients (15 per group) were included. The load of adhering P. gingivalis on the healing screw were quantified by quantitative Polymerase Chain Reaction (qPCR) Taq-Man. Immunohistochemical analysis was carried out on the gingival biopsy. Moreover, clinical data were recorded. Analysis of variance and the Holm-Sidak test was used to evaluate differences between groups. The results showed a significant low presence of P. gingivalis load in healing abutments belonging to the 0.20% CHX group. Overall, the differences in terms of P. gingivalis DNA copy number between two groups were statistically significant (p < 0.01). All implants showed very low plaque and bleeding scores, but the placebo group appeared to have the highest expression of inflammation markers for T Lymphocytes, B Lymphocytes and macrophages Cluster definitions (CD3, CD20 and CD68). The use of 0.20% CHX could be recommended in all clinical procedures as it reduces significantly P. gingivalis load and host inflammatory response around implants.

Efficacy of carvacrol against resistant rapidly growing mycobacteria in the planktonic and biofilm growth mode.

Rapidly growing mycobacteria (RGM) are environmental bacteria found worldwide with a propensity to produce skin and soft-tissue infections. Among them, the most clinically relevant species is Mycobacterium abscessus. Multiple resistance to antibiotics and the ability to form biofilm contributes considerably to the treatment failure. The search of novel anti-mycobacterial agents for the control of biofilm growth mode is crucial. The aim of the present study was to evaluate the activity of carvacrol (CAR) against planktonic and biofilm cells of resistant RGM strains. The susceptibility of RGM strains (n = 11) to antibiotics and CAR was assessed by MIC/MBC evaluation. The CAR activity was estimated by also vapour contact assay. The effect on biofilm formation and preformed biofilm was measured by evaluation of bacterial growth, biofilm biomass and biofilm metabolic activity. MIC values were equal to 64 µg/mL for most of RGM isolates (32-512 µg/mL), MBCs were 2-4 times higher than MICs, and MICs of vapours were lower (16 µg/mL for most RGM isolates) than MICs in liquid phase. Regarding the biofilm, CAR at concentrations of 1/2 × MIC and 1/4 × MIC showed a strong inhibition of biofilm formation (61-77%) and at concentration above the MIC (2-8 × MIC) produced significant inhibition of 4- and 8-day preformed biofilms. In conclusion, CAR could have a potential use, also in vapour phase, for the control of RGM.

Pistacia vera L. oleoresin and levofloxacin is a synergistic combination against resistant Helicobacter pylori strains.

The increasing multidrug resistance in Helicobacter pylori, also correlated to its biofilm-forming ability, underlines the need to search novel strategies to improve the eradication rate. Natural compounds are proposed as antibiotic-resistant-breakers capable to restore the efficacy of conventional drugs. Aim of this work was to evaluate the capability of Pistacia vera L. oleoresin (ORS) to synergize with levofloxacin (LVX) against resistant H. pylori strains. The antimicrobial activity of P. vera L. ORS and LVX and their combinations was determined by MIC/MBC (in neutral and acidic environments) and checkerboard tests. The anti-biofilm effect was determined by biomass quantification. In vivo Galleria mellonella model was used to confirm in vitro data. Pistacia vera L. ORS and LVX MICs ranged respectively from 780 to 3120 mg/l and from 0.12 to 2.00 mg/l, at pH 7.0 and 5.5. MBCs were similar to MICs. Pistacia vera L. ORS was able to synergize with LVX, restoring its effectiveness in LVX resistant strains. Pistacia vera L. ORS, LVX and their synergistic combinations displayed significant biofilm reduction. Pistacia vera L. ORS and LVX, showed protective effect against H. pylori infection on G. mellonella (62% and 63% of survival, respectively). Pistacia vera L. ORS can be considered a promising potentiator to restore the effectiveness of LVX tackling the H. pylori antibiotic resistance phenomenon.

Bovine lactoferrin enhances the efficacy of levofloxacin-based triple therapy as first-line treatment of Helicobacter pylori infection: an in vitro and in vivo study.

OBJECTIVES: To evaluate the in vitro antimicrobial/antivirulence action of bovine lactoferrin and its ability to synergize with levofloxacin against resistant Helicobacter pylori strains and to analyse the effect of levofloxacin, amoxicillin and esomeprazole with and without bovine lactoferrin as the first-line treatment for H. pylori infection. METHODS: The bovine lactoferrin antimicrobial/antivirulence effect was analysed in vitro by MIC/MBC determination and twitching motility against six clinical H. pylori strains and a reference strain. The synergism was evaluated using the chequerboard assay. The prospective therapeutic trial was carried out on two separate patient groups, one treated with esomeprazole/amoxicillin/levofloxacin and the other with esomeprazole/amoxicillin/levofloxacin/bovine lactoferrin. Treatment outcome was determined with the [13C]urea breath test. RESULTS: In vitro, bovine lactoferrin inhibited the growth of 50% of strains at 10 mg/mL and expressed 50% bactericidal effect at 40 mg/mL. The combination of levofloxacin and bovine lactoferrin displayed a synergistic effect for all strains, with the best MIC reduction of 16- and 32-fold for levofloxacin and bovine lactoferrin, respectively. Bovine lactoferrin at one-fourth MIC reduced microbial motility significantly for all strains studied. In the in vivo study, 6 of 24 patients recruited had treatment failure recorded with esomeprazole/amoxicillin/levofloxacin (75% success, 95% CI 57.68%-92.32%), and in the group with esomeprazole/amoxicillin/levofloxacin/bovine lactoferrin, 2 out of 53 patients recruited had failure recorded (96.07% success, 95% CI 90.62%-101.38%). CONCLUSIONS: Bovine lactoferrin can be considered a novel potentiator for restoring susceptibility in resistant H. pylori strains. Bovine lactoferrin added to a triple therapy in first-line treatment potentiates the therapeutic effect.

Antimicrobial and Antibiofilm Efficacy of Graphene Oxide against Chronic Wound Microorganisms.

Chronic wounds represent an increasing problem worldwide. Graphene oxide (GO) has been reported to exhibit strong antibacterial activity toward both Gram-positive and Gram-negative bacteria. The aim of this work was to investigate the in vitro antimicrobial and antibiofilm efficacy of GO against wound pathogens. Staphylococcus aureus PECHA 10, Pseudomonas aeruginosa PECHA 4, and Candida albicans X3 clinical isolates were incubated with 50 mg/liter of GO for 2 and 24 h to evaluate the antimicrobial effect. Optical and atomic force microscopy images were performed to visualize the effect of GO on microbial cells. Moreover, the antibiofilm effect of GO was tested on biofilms, both in formation and mature. Compared to the respective time controls, GO significantly reduced the S. aureus growth both at 2 and 24 h in a time-dependent way, and it displayed a bacteriostatic effect in respect to the GO t = 0; an immediate (after 2 h) slowdown of bacterial growth was detected for P. aeruginosa, whereas a tardive effect (after 24 h) was recorded for C. albicans Atomic force microscopy images showed the complete wrapping of S. aureus and C. albicans with GO sheets, which explains its antimicrobial activity. Moreover, significant inhibition of biofilm formation and a reduction of mature biofilm were recorded for each detected microorganism. The antibacterial and antibiofilm properties of GO against chronic wound microorganisms make it an interesting candidate to incorporate into wound bandages to treat and/or prevent microbial infections.

Curcumin, an antibiotic resistance breaker against a multiresistant clinical isolate of Mycobacterium abscessus.

Curcumin, a phenolic compound extracted from Curcuma longa, exerts multiple pharmacological effects, including an antimicrobial action. Mycobacterium abscessus, an environmental, nontuberculous, rapidly growing mycobacterium, is an emerging human pathogen causing serious lung infections and one of the most difficult to treat, due to its multidrug resistance and biofilm-forming ability. We wanted to evaluate the antimicrobial and antivirulence activity of curcumin and its ability to synergize with antibiotics against a clinical M. abscessus strain (29904), isolated from the bronchoaspirate of a 66-year-old woman admitted to hospital for suspected tuberculosis. Curcumin [minimum inhibitory concentrations (MIC) = 128 mg/L] was synergic (fractional inhibitory concentration index ≤0.5) with amikacin, clarithromycin, ciprofloxacin, and linezolid, to which strain 29904 showed resistance/intermediate susceptibility. Curcumin at 1/8 × MIC significantly reduced motility, whereas at 4 × MIC, it completely inhibited 4- and 8-day mature biofilms. Synergistic combinations of curcumin and amikacin induced a general reduction in microbial aggregates and substantial loss in cell viability. Disruption of 4- and 8-day biofilms was the main effect detected when curcumin was the predominant compound. The present findings support previous evidence that curcumin is a potential antibiotic resistance breaker. Curcumin, either alone or combined with antibiotics, could provide a novel strategy to combat antibiotic resistance and virulence of M. abscessus.