Log CFU Reduction Research Articles

Photodynamic inactivation of E. coli with cationic imidazolyl-porphyrin photosensitizers and their synergic combination with antimicrobial cinnamaldehyde

Bacterial infections are a global health concern, particularly due to the increasing resistance of bacteria to antibiotics. Multi-drug resistance (MDR) is a considerable challenge, and novel approaches are needed to treat bacterial infections. Photodynamic inactivation (PDI) of microorganisms is increasingly recognized as an effective method to inactivate a broad spectrum of bacteria and overcome resistance mechanisms. This study presents the synthesis of a new cationic 5,15-di-imidazolyl porphyrin derivative and the impact of n-octanol/water partition coefficient (logP) values of this class of photosensitizers on PDI efficacy of Escherichia coli. The derivative with logP = –0.5, IP-H-OH2+, achieved a remarkable 3 log CFU reduction of E. coli at 100 nM with only 1.36 J/cm2 light dose at 415 nm, twice as effective as the second-best porphyrin IP-H-Me2+, of logP = –1.35. We relate the rapid uptake of IP-H-OH2+ by E. coli to improved PDI and the very low uptake of a fluorinated derivative, IP-H-CF32+, logP ≈ 1, to its poor performance. Combination of PDI with cinnamaldehyde, a major component of the cinnamon plant known to alter bacteria cell membranes, offered synergic inactivation of E. coli (7 log CFU reduction), using 50 nM of IP-H-OH2+ and just 1.36 J/cm2 light dose. The success of combining PDI with this natural compound broadens the scope of therapies for MDR infections that do not add drug resistance. In vivo studies on a mouse model of wound infection showed the potential of cationic 5,15-di-imidazolyl porphyrins to treat clinically relevant infected wounds.Graphical

Charge adaptive phytochemical-based nanoparticles for eradication of methicillin-resistant staphylococcus aureus biofilms

The intrinsic resistance of MRSA coupled with biofilm antibiotic tolerance challenges the antibiotic treatment of MRSA biofilm infections. Phytochemical-based nanoplatform is a promising emerging approach for treatment of biofilm infection. However, their therapeutic efficacy was restricted by the low drug loading capacity and lack of selectivity. Herein, we constructed a surface charge adaptive phytochemical-based nanoparticle with high isoliquiritigenin (ISL) loading content for effective treatment of MRSA biofilm. A dimeric ISL prodrug (ISL-G2) bearing a lipase responsive ester bond was synthesized, and then encapsulated into the amphiphilic quaternized oligochitosan. The obtained ISL-G2 loaded NPs possessed positively charged surface, which allowed cis-aconityl-d-tyrosine (CA-Tyr) binding via electrostatic interaction to obtain ISL-G2@TMDCOS-Tyr NPs. The NPs maintained their negatively charged surface, thus prolonging the blood circulation time. In response to low pH in the biofilms, the fast removal of CA-Tyr led to a shift in their surface charge from negative to positive, which enhanced the accumulation and penetration of NPs in the biofilms. Sequentially, the pH-triggered release of d-tyrosine dispersed the biofilm and lipase-triggered released of ISL effectively kill biofilm MRSA. An in vivo study was performed on a MRSA biofilm infected wound model. This phytochemical-based system led to ∼2 log CFU (>99 %) reduction of biofilm MRSA as compared to untreated wound (P < 0.001) with negligible biotoxicity in mice. This phytochemical dimer nanoplatform shows great potential for long-term treatment of resistant bacterial infections.

Defining the pharmacokinetic/pharmacodynamic index of piperacillin/tazobactam within a hollow-fibre infection model to determine target attainment in intensive care patients.

It is important to optimize dosing schemes of antibiotics to maximize the probability of therapeutic success. The recommended pharmacokinetic/pharmacodynamic (PK/PD) index for piperacillin/tazobactam therapy in clinical studies ranges widely (50%-100% fT>1-4×MIC). Dosing schemes failing to achieve PK/PD targets may lead to negative treatment outcomes. The first aim of this study was to define the optimal PK/PD index of piperacillin/tazobactam with a hollow-fibre infection model (HFIM). The second aim was to predict whether these PK/PD targets are currently achieved in critically ill patients through PK/PD model simulation. A dose-fractionation study comprising 21 HFIM experiments was performed against a range of Gram-negative bacterial pathogens, doses and infusion times. Clinical data and dose histories from a case series of nine patients with a known bacterial infection treated with piperacillin/tazobactam in the ICU were collected. The PK/PD index and predicted plasma concentrations and therefore target attainment of the patients were simulated using R version 4.2.1. fT >MIC was found to be the best-fitting PK/PD index for piperacillin/tazobactam. Bactericidal activity with 2 log10 cfu reduction was associated with 77% fT>MIC. Piperacillin/tazobactam therapy was defined as clinically 'ineffective' in ∼78% (7/9) patients. Around seventy-one percent (5/7) of these patients had a probability of >10% that 2 log10 cfu reduction was not attained. Our dose-fractionation study indicates an optimal PK/PD target in piperacillin/tazobactam therapies should be 77% fT>MIC for 2 log10 kill. Doses to achieve this target should be considered when treating patients in ICU.

Edible antimicrobial yeast-based coating with basil essential oil for enhanced food safety

Foodborne pathogen contamination is recognized as a critical public health burden. The application of conventional antimicrobial strategies is significantly restricted in the food industry due to concerns over potential impacts on food quality and human health safety. Herein, this study developed a scalable methodology for encapsulating basil essential oil (BO) into yeast cell wall particles (YCWPs), creating edible antimicrobial coating suspensions for effective pathogen control on real food matrices. Thermal stability and long-term durability of the antimicrobial BO were enhanced after being encapsulated into the YCWPs, which served as a protective barrier, reducing the evaporation of small molecules in the BO mixture. In addition, benefiting from the specific affinity between the YCWP microcarriers and pathogens, the encapsulated BO with a high local concentration around the pathogens could avoid the unexpected consumption caused by organic pollutants encountered in the food system. The obtained BO@YCWPs demonstrated promising antimicrobial efficiency, even under high chemical oxygen demand (COD) conditions, achieving approximately 6 log CFU reductions of S. aureus and E. coli within 30 and 15 min, respectively. Moreover, the antimicrobial BO@YCWPs were suspended in an aqueous solution and then spray-coated onto the fresh basil leaves, significantly inhibiting the proliferation of various microorganisms and demonstrating the great potential application of the BO@YCWPs in ensuring food safety.

Development, characterization, and evaluation of a simple polymicrobial colony biofilm model for testing of antimicrobial wound dressings.

Simple biofilm models are the first step to testing of any antimicrobial and wound dressing; therefore, the aim of this study was to develop and validate a simple polymicrobial colony biofilm wound model comprised of Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans on RPMI-1640 agar. The model was then used to evaluate the topical disinfectant chlorohexidine and four commercially available wound dressings using the polymicrobial model. The model used was as a starting point to mimic debridement in clinical care of wounds and the effectiveness of wound dressings evaluated afterwards. Planktonic assessment using AATCC100-2004 demonstrated that all antimicrobial wound dressings reduced the planktonic microbial burden below the limit of detection; however, when challenged with polymicrobial colony biofilms, silver wound dressings showed limited effectiveness (1-2 log CFU reductions). In contrast, a single iodine releasing wound dressing showed potent antibiofilm activity reducing all species CFUs below the limit of detection (>6-10 log) depending on the species. A disrupted biofilm model challenge was performed to represent the debridement of a wound and wound silver-based wound dressings were found to be marginally more effective than in whole colony biofilm challenges while the iodine containing wound dressing reduced microbial recovery below the limit of detection. In this model, silver dressings were ineffective versus the whole colony biofilms but showed some recovery of activity versus the disrupted colony biofilm. The iodine wound dressing reduced the viability of all species below the level of detection. This suggests that mode of action of wound dressing should be considered for the type of biofilm challenge as should the clinical use, e.g. debridement.

Glycogen plays a key role in survival of Salmonella Typhimurium on dry surfaces and in low-moisture foods

Salmonella enterica is known to survive in desiccate environments and is often associated with low-moisture foods (LMFs). In this work, S. Typhimurium ATCC 14028 was found to survive better by achieving the least reductions (3.17 ± 0.20 Log CFU reduction) compared to S. Tennessee ATCC 10722 (3.82 ± 0.13 Log CFU reduction) and S. Newport ATCC 6962 (6.03 ± 0.36 Log CFU reduction) after 30 days on surfaces with a relative humidity of 49% at ambient temperature. A metabolomic analysis revealed that S. Typhimurium was still active in energy metabolism after 24 h in the desiccate environment and glycogen, an energy reserve, was drastically reduced. We followed up on the glycogen levels over 30 days and found indeed a sharp decline on the first day. However, the glycogens detected on day 7 were significantly higher (P < 0.05) and thereafter remained stable above the original levels until day 30. The expression levels of both glycogen anabolism- and catabolism-related genes (csrA, glgA, glgC, glgX) were significantly up-regulated at all tested points (P < 0.05). The glgA and glgC insertion mutants displayed weaker survivability on both dry surfaces and in representative LMFs (flour and milk powder) compared to the wild-type strain. This work highlights the role of glycogen during different periods of desiccation, which may bring novel insight into mitigating Salmonella by disrupting glycogen metabolism.

Plasma-activated water for inactivation of Salmonella Typhimurium avirulent surrogate: Applications in produce and shell egg and understanding the modes of action

Plasma-activated water (PAW) is gaining interest as a novel sanitizer for foods. However, the sanitizing efficacy of PAW varies under different conditions, and its modes of action and impacts on food quality need further investigation for it to be applied in the food industry. This study evaluated the inactivation efficacy of PAW against a Salmonella Typhimurium avirulent surrogate, investigated the underlying mechanisms, and assessed the impacts of PAW on the safety and quality of cherry tomatoes and shell eggs. PAW was effective in inactivating S. Typhimurium in a planktonic state (1.5–5.8 log CFU reduction in 2–5 min) and on the food surface (>5 log CFU reduction in 5 min). PAW prepared with deionized water vs. tap water did not differ significantly in sanitizing efficacy. Various reactive oxygen and nitrogen species (RONS) contributed to the antimicrobial property of PAW, with nitric oxide playing the most significant role. The acidic nature of PAW might synergistically affect bacteria by disrupting cell membrane and enhancing RONS permeation. PAW did not modify the visual appearance and internal pH of the treated foods. This study demonstrates that PAW has the potential to be applied as a novel sanitizer for fresh produce and egg washing.

Storage-related stability and antimicrobial efficacy of bottled, neutral-pH Electrolysed Oxidising Water

ObjectivesDenture stomatitis is prevalent in older people and poses serious health risks. Ready-to-use (RTU) neutral-pH Electrolysed Oxidizing Water (EOW) is an effective environmental disinfectant used in residential care settings and geriatric wards. However, the influence of storage on stability and effectiveness for denture disinfection has not been established. This research investigated the storage-related stability and antimicrobial activity of RTU EOW, and its efficacy against Candida albicans biofilms formed on denture resin. MethodsThe pH, oxidation/reduction potential (mV), available chlorine content (mg/L) and [HOCl] (mM) of RTU EOW (Envirolyte, New Zealand) solutions (n = 22) were measured from bottle opening to 28 days following storage at 4 °C, room temperature (RT) or 37 °C. Staphylococcus aureus and C. albicans cells were incubated in 80% EOW for contact times (CTs) up to 15 min and colony-forming units (cfu) determined. Minimum inhibitory concentrations (MIC90 EOW-HOCl) after CTs up to five minutes were determined for S. aureus and C. albicans reference strains and clinical isolates. C. albicans-denture resin disc biofilms were assessed after a five-minute CT with undiluted EOW by XTT-metabolic activity assay. Results[HOCl] remained stable when RTU EOW was stored at 4 °C or RT for five months after manufacture. One-minute CT resulted in log10 cfu reductions of >6 for S. aureus and >5 for C. albicans. Mean MIC90 for five-minute CT was 37 µM (S. aureus) and 54 µM (C. albicans). Undiluted EOW reduced C. albicans biofilm metabolic activity by 86%. ConclusionsRTU neutral-pH EOW is stable over five-months storage and is an effective denture disinfectant. Clinical significanceThe efficacy of the RTU neutral EOW against C. albicans isolates and biofilms formed on denture resin surfaces supports its use as a denture disinfectant and can inform future research to assess its potential for preventing denture-related oral Candida infections in the older population, especially in resource-limited communities.

Dielectric and dynamic antibacterial investigations of organic–inorganic conductive membranes based on oxidized cellulose with BNKT nanoceramics

The development of eco-friendly materials for advanced applications is highly demanded. The current study focuses on the preparation of conductive membranes based on tricarboxylic cellulose (TCC) loaded with cubic bismuth sodium titanate (BNKT) nanoceramics. FTIR, SEM, and EDX analyses confirm the presence of loaded BNKT on the membranes. The electrical response of the cellulose/xBNKT (x = 5, 10, 15, and 20% wt/wt) membrane is investigated using impedance spectroscopy. The real part (Z′) and the imaginary part (Z″) of the complex impedance are studied as a function of frequency (4Hz ~ 8MHz) and temperature (20 ~ 160 °C) for the different compositions. Impedance and modulus studies reveal a Debye-type relaxation phenomenon. The dielectric studies manifest promising dielectric properties. The bactericidal performance of all nanomembranes is evaluated. The nanomembrane with 20% BNKT (C20) exhibits bactericidal activity against Gram-negative and Gram-positive bacteria, with 6 log CFU reductions observed after an exposure time of 180 min. Treatment with the C20 nanomembrane shows the highest amounts of protein efflux. The results indicate that the C20 nanomembrane layer eradicated all bacterial cells. The findings suggest that the C20 nanomembrane is recommended as an intelligent and innovative antibacterial nano-system for bio-applications.

Exploring Washing Procedures for Produce Brush Washer

Previous environmental monitoring projects in food production facilities have revealed inconsistencies in how produce brush washer machines are cleaned after use; thus, the study of effective sanitation procedures for these machines is needed. Four chlorine solution treatments (ranging from 25 to 200 ppm), as well as a water-only treatment, were tested for efficacy in reducing bacterial loads for a selected small brush washer machine. Results indicate that rinsing with the machine’s power and water alone, a frequent practice among some produce processors, yielded a reduction of 0.91–1.96 log CFU per brush roller in bacterial counts, which was not statistically significant (p > 0.05). However, the chlorine treatments were found to be effective in reducing bacterial loads significantly, with higher concentrations being the most effective. The 200 ppm and 100 ppm chlorine treatments yielded bacterial reductions of 4.08 and 3.95 log CFU per brush roller, respectively, leaving bacterial levels statistically similar to the levels at postprocess decontamination, meaning these are the most effective at killing bacteria of all the chlorine concentrations tested. These data suggest the use of at least 100 ppm chlorine sanitizer solution is a good method to sanitize hard-to-clean produce washing machines, yielding an approximate 4 log CFU reduction of the inoculated bacteria.

Novel fluorobenzothiazole as a dual inhibitor of gyrase B and topoisomerase IV against Gram-positive pathogens.

Aim: The development of a novel inhibitor targeting gyrase B and topoisomerase IV offers an opportunity to combat multidrug resistance. Methods: We investigated the activity of RBx 10080758 against Gram-positive bacteria in vitro and in vivo. Results: RBx 10080758 showed a potent 50% inhibitory concentration of 0.13μM and 0.25μM against gyrase B and topoisomerase IV, respectively, and exhibited strong whole-cell in vitro activity with MIC ranges of 0.015-0.06 and 0.015-0.03μg/ml against Staphylococcus aureus and Streptococcus pneumoniae, respectively. In a rat thigh infection model with methicillin-resistant S. aureus, RBx 10080758 at 45mg/kg exhibited a>3 log10 CFU reduction in thigh muscles. Conclusion: RBx 10080758 displayed potent activity against multiple multidrug-resistantGram-positive bacteria with a dual-targeting mechanism of action.

EPS9.07 Improving colistin activity against Pseudomonas aeruginosa biofilms

Colistin is the last-resort therapeutic option in infections by multi-drug resistant pathogens and for this it has been included by World Health Organization among the “highest priority critically important antimicrobials.” Colistin binding to outer membrane (OM) of Gram negative bacteria leads to membrane destabilization, increasing permeability and finally cell death. OM remodeling, by chemical groups that reduce its negative charge, induces resistance to colistin. In Pseudomonas aeruginosa addition of aminoarabinosyl residues (Ara4N) to the lipid-A is the major pathway leading to colistin resistance, whose last step is catalyzed by the aminoarabinosyl transferase ArnT. We have previously identified a natural compound, BBN149 (renamed FDO), as putative inhibitors of ArnT, which potentiates colistin activity against planktonic colistin-resistant P aeruginosa strain (Ghirga et al., 2020, doi:10.1093/jac/dkaa200). As biofilms are relevant to the persistence of bacterial infection, we tested the colistin adjuvant activity of FDO, and its derivative FDO-H, on P. aeruginosabiofilms. First, biofilms were grown in medium containing colistin, and the resulting biofilms have been quantified by crystal violet, metabolic assay and viable cell counting. The resulting data showed a dose-dependent reduction in metabolic activity, total biomass and cell viability of biofilms treated with colistin. In biofilms treated with colistin and FDO or FDO-H, viable cells counting showed the adjuvant activity of both compounds with one log reduction of CFU in sample treated with colistin +compound respect to colistin only. Importantly, similar results were obtained with an in vitro evolved P. aeruginosa reference strain (PA14 colR5, doi:10.1128/AAC.01820-17) and with a clinical colR isolate. Collectively, our data suggest that FDO and FDO-H potentiate colistin activity against P. aeruginosa biofilms.

Effect of Food Matrix and Treatment Time on the Effectiveness of Grape Seed Extract as an Antilisterial Treatment in Fresh Produce.

Listeriosis outbreaks were associated with contaminated fruits and vegetables, including cantaloupe, apples, and celery. Grape seed extract (GSE) is a natural antimicrobial with potential for reducing Listeria monocytogenes contamination in food. This study assessed the effectiveness of GSE to reduce L. monocytogenes on fresh produce and the impact of food matrices on its antilisterial activity. GSE showed MIC values of 30-35 μg/mL against four Listeria strains used in this study. A total of 100 g portions of cantaloupe, apples, and celery were inoculated with L. monocytogenes and treated with 100-1000 μg/mL of GSE for 5 or 15 min. Results were analyzed using Rstudio and a Tukey's test. Treated produce had significantly lower L. monocytogenes counts than the control samples (p-value < 0.05). The inhibition was significantly higher on apples and lowest on cantaloupe. Moreover, a 15 min treatment was found to be more effective than a 5 min treatment in reducing L. monocytogenes on all produce types. The reduction in L. monocytogenes levels varied between 0.61 and 2.5 log10 CFU reduction, depending on the treatment concentration, duration, and produce matrix. These findings suggest that GSE is an effective antilisterial treatment for fresh produce, with varying levels of effectiveness depending on the food matrix and treatment time.

Efficacy of PBTZ169 and pretomanid against Mycobacterium avium, Mycobacterium abscessus, Mycobacterium chelonae, and Mycobacterium fortuitum in BALB/c mice models.

We aimed to evaluate the activity of PBTZ169 and pretomanid against non-tuberculous mycobacteriosis (NTM) in vitro and in vivo. The minimum inhibitory concentrations (MICs) of 11 antibiotics, against slow-growing mycobacteria (SGMs) and rapid-growing mycobacteria (RGMs) were tested using the microplate alamarBlue assay. The in vivo activities of bedaquiline, clofazimine, moxifloxacin, rifabutin, PBTZ169 and pretomanid against four common NTMs were assessed in murine models. PBTZ169 and pretomanid had MICs of >32 μg/mL against most NTM reference and clinical strains. However, PBTZ169 was bactericidal against Mycobacterium abscessus (3.33 and 1.49 log10 CFU reductions in the lungs and spleen, respectively) and Mycobacterium chelonae (2.29 and 2.24 CFU reductions in the lungs and spleen, respectively) in mice, and bacteriostatic against Mycobacterium avium and Mycobacterium fortuitum. Pretomanid dramatically decreased the CFU counts of M. abscessus (3.12 and 2.30 log10 CFU reductions in the lungs and spleen, respectively), whereas it showed moderate inhibition of M. chelonae and M. fortuitum. Bedaquiline, clofazimine, and moxifloxacin showed good activities against four NTMs in vitro and in vivo. Rifabutin did not inhibit M. avium and M. abscessus in mice. PBTZ169 appears to be a candidate for treating four common NTM infections. Pretomanid was more active against M. abscessus, M. chelonae and M. fortuitum than against M. avium.

Photoantibacterial Poly(vinyl)chloride Films Applying Curcumin Derivatives as Bio-Based Plasticizers and Photosensitizers.

Herein we describe the design of natural curcumin ester and ether derivatives and their application as potential bioplasticizers, to prepare photosensitive phthalate-free PVC-based materials. The preparation of PVC-based films incorporating several loadings of newly synthesized curcumin derivatives along with their standard solid-state characterization is also described. Remarkably, the plasticizing effect of the curcumin derivatives in the PVC material was found to be similar to that observed in previous PVC-phthalate materials. Finally, studies applying these new materials in the photoinactivation of S. aureus planktonic cultures revealed a strong structure/activity correlation, with the photosensitive materials reaching up to 6 log CFU reduction at low irradiation intensities.

Phage-Based Control of Methicillin Resistant Staphylococcus aureus in a Galleria mellonella Model of Implant-Associated Infection

Staphylococcus aureus implant-associated infections are difficult to treat because of the ability of bacteria to form biofilm on medical devices. Here, the efficacy of Sb-1 to control or prevent S. aureus colonization on medical foreign bodies was investigated in a Galleria mellonella larval infection model. For colonization control assays, sterile K-wires were implanted into larva prolegs. After 2 days, larvae were infected with methicillin-resistant S. aureus ATCC 43300 and incubated at 37 °C for a further 2 days, when treatments with either daptomycin (4 mg/kg), Sb-1 (107 PFUs) or a combination of them (3 x/day) were started. For biofilm prevention assays, larvae were pre-treated with either vancomycin (10 mg/kg) or Sb-1 (107 PFUs) before the S. aureus infection. In both experimental settings, K-wires were explanted for colony counting two days after treatment. In comparison to the untreated control, more than a 4 log10 CFU and 1 log10 CFU reduction was observed on K-wires recovered from larvae treated with the Sb-1/daptomycin combination and with their singular administration, respectively. Moreover, pre-infection treatment with Sb-1 was found to prevent K-wire colonization, similarly to vancomycin. Taken together, the obtained results demonstrated the strong potential of the Sb-1 antibiotic combinatory administration or the Sb-1 pretreatment to control or prevent S. aureus-associated implant infections.

Anti-Salmonella polyvinyl alcohol coating containing a virulent phage PBSE191 and its application on chicken eggshell

The use of antibiotics in the food industry is avoided due to the increase of antibiotic-resistant bacteria. Therefore, the bacteriophage is emerging as an alternative agent. Here, we characterized the Salmonella Enteritidis phage PBSE191 and applied it to a polyvinyl alcohol (PVA) film. Transmission electron microscopic analysis revealed that it belonged to the Caudoviricetes class, with an icosahedral head and flexible tails. The phage showed rapid and strong lytic activity within 1 h. It was active against a broad range of Salmonella isolates, including six serotypes. In 25 min, 99 % of the initial population was adsorbed to the bacterial cell surface. The phage was also applied to 10 % (w/v) PVA films and coatings, which were then characterized in terms of phage stability and antibacterial performance, both in vitro and in foods. The phage remained stable in the 10 % (w/v) PVA solution containing 20 % (w/w, based on PVA weight) sorbitol (PVAS20), indicating that the phage was stable under dry conditions and strongly released in the polymer. Furthermore, significant bacterial cell reduction (2.0 × 105 CFU/film within 2 h) was observed in the phage-containing PVAS20 films. In addition, the PBSE191-containing PVAS20 coating on the chicken eggshell surface showed significant anti-Salmonella efficiency (about 2 log CFU reduction) within 24 h. Overall, the PBSE191 phage possesses a high potential as a biocontrol agent for use as an additive, or as an active antibacterial packaging to improve food safety against Salmonella contamination.

Antioxidant, Anti-Inflammatory, and Antibacterial Properties of an Achillea millefolium L. Extract and Its Fractions Obtained by Supercritical Anti-Solvent Fractionation against Helicobacter pylori.

The main objective of this work is to evaluate the potential utility of an Achillea millefolium extract (yarrow extract, YE) in the control of H. pylori infection. The supercritical anti-solvent fractionation (SAF) process of YE allowed the obtaining of two different fractions: yarrow’s precipitated fraction (YPF), enriched in most polar phenolic compounds (luteolin-7-O-glucoside, luteolin, and 3,5-dicaffeoylquinic acid), and yarrow’s separator fraction (YSF), enriched in monoterpenes and sesquiterpenes, mainly containing camphor, artemisia ketone, and borneol. YE was effective in reducing reactive oxygen species (ROS) production in human gastric AGS cells by 16% to 29%, depending on the H. pylori strain. YPF had the highest inhibitory activity (38–40%) for ROS production. YE modulated the inflammatory response in AGS gastric cells, decreasing IL-8 production by 53% to 64%. This IL-8 inhibition also showed a strain-dependent character. YPF and YSF exhibited similar behavior, reducing IL-8 production, suggesting that both phenolic compounds and essential oils could contribute to IL-8 inhibition. YSF showed the highest antibacterial activity against H. pylori (6.3–7.1 log CFU reduction, depending on the strain) and lower MIC (0.08 mg/mL). Results obtained have shown that YE and SAF fractions (YPF and YSF) were effective as antioxidant, anti-inflammatory, and antibacterial agents regardless of the H. pylori strain characteristics.

Synergy between Mecillinam and Ceftazidime/Avibactam or Avibactam against Multi-Drug-Resistant Carbapenemase-Producing Escherichia coli and Klebsiella pneumoniae.

Background: Carbapenemase-producing Klebsiella pneumoniae and Escherichia coli have become a significant global health challenge. This has created an urgent need for new treatment modalities. We evaluated the efficacy of mecillinam in combination with either avibactam or ceftazidime/avibactam against carbapenemase-producing clinical isolates. Materials and methods: Nineteen MDR clinical isolates of K. pneumoniae and E. coli were selected for the presence of blaKPC, blaNDM, blaOXA or blaIMP based on whole-genome sequencing and phenotypic susceptibility testing. We tested the synergy between mecillinam and avibactam or ceftazidime/avibactam. We used time–kill studies in vitro and a mouse peritonitis/sepsis model to confirm the synergistic effect. We investigated avibactam’s impact on mecillinam´s affinity for penicillin-binding proteins with a Bocillin assay, and cell changes with phase-contrast and confocal laser scanning microscopy. Results: Mecillinam combined with ceftazidime/avibactam or avibactam substantially reduced MICs (from up to >256 µg/mL to <0.0016 µg/mL) for 17/18 strains. Significant log-CFU reductions were confirmed in time–kill and in vivo experiments. The Bocillin assay did not reveal changes. Conclusion: Mecillinam in combination with avibactam or ceftazidime/avibactam has a notable effect on most types of CPEs, both in vitro and in vivo. The mecillinam/avibactam combination treatment could be a new efficient antibiotic treatment against multi-drug-resistant carbapenemase-producing Gram-negative pathogens.

Trimethoprim/sulfamethoxazole pharmacodynamics against Stenotrophomonas maltophilia in the in vitro chemostat model.

Trimethoprim/sulfamethoxazole has historically been the treatment of choice for infection caused by Stenotrophomonas maltophilia. This study sought to define the pharmacodynamic indices and magnitude of exposure required for stasis and 1 log10 cfu reductions. Pharmacodynamic studies were conducted using the in vitro chemostat model over 24 h against three trimethoprim/sulfamethoxazole-susceptible S. maltophilia isolates with MICs from 0.25/4.75 to 2/38 mg/L. The primary endpoint was the change in cfu at 24 h relative to baseline. The log ratio of the area under the cfu curve (LR AUcfu) was a secondary endpoint. Trimethoprim and sulfamethoxazole exposures required for stasis and 1 log10 cfu/mL reduction were determined. Trimethoprim/sulfamethoxazole exposures achieved stasis and 1 log10 cfu/mL reductions in 9/16 (56%) and 2/16 (13%) of experiments. Both the fAUC/MIC and fCmax/MIC were identified as equivalent pharmacodynamic drivers, with stasis achieved at an fAUC/MIC of 67.4 and 30.0 for trimethoprim and sulfamethoxazole, respectively. Clinically meaningful exposures required to achieve 1 log10 cfu/mL reductions were not quantifiable. The LR AUcfu analysis supported the lack of overall bacterial burden reduction against S. maltophilia. In this in vitro chemostat model, trimethoprim/sulfamethoxazole monotherapy, even at higher doses, achieved limited activity against susceptible S. maltophilia.