Bacterial Regrowth Research Articles

In vitro Optimization of Ceftazidime/Avibactam for KPC-Producing Klebsiella pneumoniae.

Ceftazidime/avibactam is an important treatment option for infections caused by Klebsiella pneumoniae carbapenemase-producing K. pneumoniae (KPC-Kp), however, resistance can emerge during treatment. The objective of the study was to define the ceftazidime/avibactam concentrations required to suppress bacterial regrowth in ceftazidime/avibactam susceptible isolates and identify active therapies against ceftazidime/avibactam-resistant KPC-Kp. Time-kill assays were performed against twelve ST258 KPC-Kp isolates that harbored blaKPC–2 or blaKPC–3. Nine KPC-Kp isolates (KPC-Kp 5A, 6A, 7A, 8A, 9A, 24A, 25A, 26A, and 27A) were susceptible to ceftazidime/avibactam, two (KPC-Kp 6B and 7B) were ceftazidime/avibactam resistant and meropenem susceptible, and one (KPC-Kp 1244) was resistant to both ceftazidime/avibactam and meropenem. Sequencing of the blaKPC genes revealed mutations in KPC-Kp 6B (D179Y substitution) and 7B (novel 21 base pair deletion) that both affected the Ω-loop encoding portion of the gene. Time-kill assays showed that against ceftazidime/avibactam-susceptible KPC-Kp, ceftazidime/avibactam concentrations ≥40/7.5 mg/L caused mean 5.42 log10CFU/mL killing and suppressed regrowth. However, regrowth occurred for some KPC-Kp isolates with a ceftazidime/avibactam concentration of 20/3.75 mg/L. Against ceftazidime/avibactam-resistant and meropenem-susceptible KPC-Kp 6B and 7B, bactericidal activity and synergy was observed for ceftazidime/avibactam in combination with meropenem ≤3.125 mg/L, while meropenem concentrations ≥50 mg/L were bactericidal as monotherapy. In contrast, clinically achievable concentrations of ceftazidime/avibactam were bactericidal against KPC-Kp 1244, which was ceftazidime/avibactam-resistant and meropenem-resistant due to outer membrane porin mutations and elevated blaKPC expression. Achieving high ceftazidime/avibactam concentrations may help to suppress bacterial regrowth in the presence of ceftazidime/avibactam. The optimal treatment approach for ceftazidime/avibactam-resistant KPC-Kp likely depends on the mechanism of resistance. Additional studies are warranted to confirm these findings.

Control of Burn Wound Infection by Methylene Blue-Mediated Photodynamic Treatment With Light-Emitting Diode Array Illumination in Rats.

Control of burn wound infection is difficult due to the increase in drug-resistant bacteria and deteriorated immune responses. In this study, we examined the usefulness of methylene blue (MB)-mediated antimicrobial photodynamic therapy (aPDT) with illumination by a light-emitting diode (LED) array for controlling invasive infections from the wound to inside the body for rats with an extended deep burn infected with Pseudomonas aeruginosa. An MB solution with the addition of ethanol, ethylene-diamine-tetra-acetic acid disodium salt, and dimethyl sulfoxide was used as a photosensitizer (PS). An extended deep burn was made on the dorsal skin in rats and the wounds were infected with P. aeruginosa. The rats were divided into three groups: control (no treatment; n = 14), PS mixture application alone (PS alone group; n = 10), and aPDT group (n = 14). For aPDT, after the PS mixture was applied onto the surface of infected wounds, the wounds were illuminated with a 665-nm LED array at an intensity of 45 mW/cm2 three times per treatment, with an illumination duration of 20 minutes and an interval of 10 minutes. The treatment was repeated each day for 7 consecutive days (day 0-day 6). Bacterial numbers on the wound surface and the weights and survival rates of the animals were evaluated daily. At the endpoints, bacterial numbers in the liver and blood were counted. Since the PS mixture showed high dark toxicity against P. aeruginosa in vitro, the influence of the PS mixture application onto healthy skin was also examined in vivo. Even in the aPDT group, rapid bacterial regrowth was observed on the wound surface after each day's treatment, but the geometric mean values of the bacterial numbers before and after each aPDT were considerably lower than those in the control group. Application of the PS mixture alone showed a clear bactericidal effect only at day 0, which is attributable to the formation of biofilms after day 1. Rats in the aPDT group showed a smaller weight loss, a higher ratio of no bacterial migration at the endpoints, and significantly higher survival rates than those in the other two groups. Effects of repeated application of the PS mixture onto healthy skin were not evident. Application of MB-mediated aPDT with illumination by a high-intensity LED array daily for seven consecutive days was effective for suppressing invasive infection from the wound to inside the body in rats with an extensive deep burn infected with P. aeruginosa, resulting in significant improvement of their survival. Lasers Surg. Med. © 2021 Wiley Periodicals LLC.

In Vitro Newly Isolated Environmental Phage Activity against Biofilms Preformed by Pseudomonas aeruginosa from Patients with Cystic Fibrosis.

As disease worsens in patients with cystic fibrosis (CF), Pseudomonas aeruginosa (PA) colonizes the lungs, causing pulmonary failure and mortality. Progressively, PA forms typical biofilms, and antibiotic treatments determine multidrug-resistant (MDR) PA strains. To advance new therapies against MDR PA, research has reappraised bacteriophages (phages), viruses naturally infecting bacteria. Because few in vitro studies have tested phages on CF PA biofilms, general reliability remains unclear. This study aimed to test in vitro newly isolated environmental phage activity against PA isolates from patients with CF at Bambino Gesù Children’s Hospital (OBG), Rome, Italy. After testing in vitro phage activities, we combined phages with amikacin, meropenem, and tobramycin against CF PA pre-formed biofilms. We also investigated new emerging morphotypes and bacterial regrowth. We obtained 22 newly isolated phages from various environments, including OBG. In about 94% of 32 CF PA isolates tested, these phages showed in vitro PA lysis. Despite poor efficacy against chronic CF PA, five selected-lytic-phages (Φ4_ZP1, Φ9_ZP2, Φ14_OBG, Φ17_OBG, and Φ19_OBG) showed wide host activity. The Φ4_ZP1-meropenem and Φ14_OBG-tobramycin combinations significantly reduced CF PA biofilms (p < 0.001). To advance potential combined phage-antibiotic therapy, we envisage further in vitro test combinations with newly isolated phages, including those from hospital environments, against CF PA biofilms from early and chronic infections.

Isolation, identification and reviewing the health effect of HPC bacteria in household point-of-use (PoU) water treatment devices: a case study, Ahvaz, Iran.

In Ahwaz, a city in west southern Iran, the majority of households are using Point of Use (PoU) water treatment units. The heterotrophic plate count (HPC) bacteria were isolated from these units while they were mounted on water distribution system in order to determine the variations in HPC and diversity of the bacterial population using polymerase chain reaction (PCR). Results showed that bacterial population regrowth in PoU units could increase HPC exceeding the limit of the 500CFU/mL in outlet water. In around 70% of the input water samples, the HPC was less than 500CFU/ml with a mean of 226.7 (CI 95%: 28.1-425.3). HPC in output treated water samples had an increasing trend from the start of the unit operation with a mean of 2416.4 (CI 95%: 1074.9-3757.9). Out of 49 detected bacterial strains, 20 strains were Gram-negative and 29 Gram-positive. Bacillus was the most frequent genes detected in inlet and outlet water samples. Most of the identified bacterial strains were opportunistic pathogens potentially dangerous for immunocompromised population. HPC population in PoU units significantly can be increased during a one-month period of operation, so replacement of the filters must be done regularly.

Modeling Bacterial Regrowth and Trihalomethane Formation in Water Distribution Systems

The formation of bacterial regrowth and disinfection by-products is ubiquitous in chlorinated water distribution systems (WDSs) operated with organic loads. A generic, easy-to-use mechanistic model describing the fundamental processes governing the interrelationship between chlorine, total organic carbon (TOC), and bacteria to analyze the spatiotemporal water quality variations in WDSs was developed using EPANET-MSX. The representation of multispecies reactions was simplified to minimize the interdependent model parameters. The physicochemical/biological processes that cannot be experimentally determined were neglected. The effects of source water characteristics and water residence time on controlling bacterial regrowth and Trihalomethane (THM) formation in two well-tested systems under chlorinated and non-chlorinated conditions were analyzed by applying the model. The results established that a 100% increase in the free chlorine concentration and a 50% reduction in the TOC at the source effectuated a 5.87 log scale decrement in the bacteriological activity at the expense of a 60% increase in THM formation. The sensitivity study showed the impact of the operating conditions and the network characteristics in determining parameter sensitivities to model outputs. The maximum specific growth rate constant for bulk phase bacteria was found to be the most sensitive parameter to the predicted bacterial regrowth.

Pharmacodynamics of once- versus twice-daily dosing of nebulized amikacin in an in vitro Hollow-Fiber Infection Model against 3 clinical isolates of Pseudomonas aeruginosa

This study aims to compare the bacterial killing of once- versus twice-daily nebulized amikacin against Pseudomonas aeruginosa and to determine the optimal duration of therapy. Three clinical P. aeruginosa isolates (amikacin MICs 2, 8, and 64 mg/L) were exposed to simulated epithelial lining fluid exposures of nebulized amikacin with dosing regimens of 400 mg and 800 mg once- or twice-daily up to 7-days using the in vitro hollow-fiber infection model. Quantitative cultures were performed. Simulated amikacin dosing regimens of 400 mg twice-daily and 800 mg once-daily achieved ≥2-log reduction in the bacterial burden within the first 24-hours of therapy for all isolates tested. No dosing regimen suppressed the emergence of amikacin resistance. No difference in bacterial killing or regrowth was observed between 3- and 7-days of amikacin. Amikacin doses of 800 mg once-daily for up to 3-days may be considered for future clinical trials.

Water Lice and Other Macroinvertebrates in Drinking Water Pipes: Diversity, Abundance and Health Risk

Activities to ensure and maintain water quality in drinking water networks, including flushing, are presented after standardized hydrant sampling combined with a stainless-steel low pressure–high flow rate (NDHF) filter and a 100 µm mesh size was used to separate pipe inhabitants. A databank of more than 1000 hydrant samples in European lowland areas was developed and used to analyze the diversity and abundance of macroinvertebrates in drinking water networks. Load classes for water louse (Asellus aquaticus) and oligochaetes are given with three evaluation classes: normal colonization, increased colonization, and mass development. The response of Asellus aquaticus in drinking water networks to environmental conditions are presented as are their growth and reproduction, promotion of a third generation by climate change effects, food limitations, and the composition and stability of their feces. Finally, the health risks posed by dead water lice and water lice feces with bacterial regrowth and the promotion of microbe development on house filters are analyzed.

An Optimized Bacteriophage Cocktail Can Effectively Control Salmonella in vitro and in Galleria mellonella.

Salmonella spp. is a leading cause of gastrointestinal enteritis in humans where it is largely contracted via contaminated poultry and pork. Phages can be used to control Salmonella infection in the animals, which could break the cycle of infection before the products are accessible for consumption. Here, the potential of 21 myoviruses and a siphovirus to eliminate Salmonella in vitro and in vivo was examined with the aim of developing a biocontrol strategy to curtail the infection in poultry and swine. Together, the phages targeted the twenty-three poultry and ten swine prevalent Salmonella serotype isolates tested. Although individual phages significantly reduced bacterial growth of representative isolates within 6 h post-infection, bacterial regrowth occurred 1 h later, indicating proliferation of resistant strains. To curtail bacteriophage resistance, a novel three-phage cocktail was developed in vitro, and further investigated in an optimized Galleria mellonella larva Salmonella infection model colonized with representative swine, chicken and laboratory strains. For all the strains examined, G. mellonella larvae given phages 2 h prior to bacterial exposure (prophylactic regimen) survived and Salmonella was undetectable 24 h post-phage treatment and throughout the experimental time (72 h). Administering phages with bacteria (co-infection), or 2 h post-bacterial exposure (remedial regimen) also improved survival (73–100% and 15–88%, respectively), but was less effective than prophylaxis application. These pre-livestock data support the future application of this cocktail for further development to effectively treat Salmonella infection in poultry and pigs. Future work will focus on cocktail formulation to ensure stability and incorporation into feeds and used to treat the infection in target animals.

A solid oxide fuel cell fuelled by methane recovered from groundwater

This study investigates the feasibility of electricity production in a solid oxide fuel cell using methane recovered from groundwater as the fuel. Methane must be removed from groundwater for the production of drinking water to, amongst others, avoid bacterial regrowth. Instead of releasing methane to the atmosphere or converting it to carbon dioxide by flaring, methane can also be recovered by vacuum stripping and served as a fuel. However, the electrical efficiency of currently used combustion-based technologies fuelled with methane-rich gas is limited to 35% due to the low heating value of the recovered gas (70 mol. % methane) and power derating due to the presence of carbon dioxide (25 mol.%). We propose to use a solid oxide fuel cell to use the methane-rich gas as fuel. Solid Oxide Fuel Cells are fuel-flexible and potentially attain higher electrical efficiencies up to 60%. To this end, specific gas processing, including cleaning and methane reforming, is required to allow for durable operation in a solid oxide fuel cell. We assessed whether electricity could be generated by a solid oxide fuel cell using methane recovered from a full-scale drinking water treatment plant as a fuel. The groundwater had a methane concentration of 45 mg∙L-1, and the recovered gas by vacuum towers contained 70 mol% methane. We used a gas cleaning reactor with impregnated activated carbon to remove hydrogen sulfide traces from the methane-rich gas. Thermodynamic calculations showed that additional steam is required to achieve a high methane reforming. The added steam and the carbon dioxide content in the recovered gas simultaneously contribute to the methane reforming to prevent carbon deposition. The measured open circuit potential corresponded with the theoretical Nernst voltage, implying high methane reforming in the solid oxide fuel cell. The achieved power density of the cell fuelled with the methane-rich gas (mixed with steam) was 27% less than the hydrogen-fuelled cell. Ultimately, 51.2% of the power demand of the plant can be covered by replacing the gas engine in a drinking water treatment with a 915 kW solid oxide fuel cell system fuelled by the methane recovered from the groundwater, while the greenhouse gas emission can be reduced by 17.6%.

Simulation and vulnerability assessment of water distribution networks under deliberate contamination attacks

ABSTRACT This work proposes a modeling framework to quantify the effects of deliberate organic contamination events on water distribution systems. A bacterial regrowth model and a first parallel chlorine decay model were combined to describe the biochemical water species kinetics during a deliberate contaminant injection consisting of organic (TOC) and bacterial loads. Several attacks were modeled using EPANET-MSX. The impact of each attack was quantified using vulnerability indexes measuring the number of consumers affected and other metrics. These vulnerability indexes were then used to depict the network’s response to a potential threat and identify the most critical areas of the network. A sensitivity analysis was also performed to determine the network’s sensitivity to three dynamic network variables (mass injection, injection duration, and organic carbon concertation in water). An increase on bacterial populations was observed during the contaminant spread to the network, which in turn signals additional health risk to consumers.

Enhancing disinfection performance of the carbon fiber-based flow-through electrode system (FES) by alternating pulse current (APC) with low-frequency square wave

This study was aimed to investigate the influence of low-frequency alternative pulse current (APC) with square wave on the disinfection performance of carbon fiber-based flow-through electrode system (FES) towards a model bacterium (Escherichia coli). The FES disinfection under direct current (DC) supply was limited by the weak disinfection efficiency of cathode. Compared with DC supply (less than 1 log removal), FES under APC supply achieved superior disinfection performance (over 6 log removal) at the voltage of 3 V and flow rate of 125 mL/min. The optimum condition was acquired when pulse cycle time (T) equaled to hydraulic retention time (HRT), i.e. HRT/T = 1, corresponding to the low frequency (less than 1 Hz), under which the disinfection performance was improved by alternating of anode and cathode. In-situ sampling experiments revealed that redox reactions were carried out alternatively in each electrode, and both electrodes made great contributions to disinfection under APC supply, which was significantly different from DC supply. In addition, FES disinfection under APC supply also caused irreversible damage to bacterial cells, resulting in no bacterial regrowth/reactivation during storage, and guaranteed low energy consumption (11.4 ~ 12.1 Wh/m3) with high flow rate (100 ~ 125 mL/min). Therefore, the development of a novel FES under APC supply provided exciting possibilities for future application of electrochemical disinfection to achieve superior disinfection performance with no bacterial regrowth and low energy consumption.

Implementation of Pre-Milking Udder Preparation to Improve Milk Quality in Small Buffalo Dairy Farm

Pre milking udder preparations play an important role in increasing quality and safety of milk. The objective of this study is to evaluate the effect of implementation of pre milking teat dipping technique by using different chemical sanitizers (Iodophore and Quaternary ammonium compounds QACs). Materials and Methods: Pre-milking udder preparation was carried out in a private small-scale buffalo dairy farm. A total number of 30 dairy buffalo, equal in their milk production and free from clinical and subclinical mastitis, were chosen for this study. They were divided into 3 groups (10 animals each). The first group was used as control group, the second group was treated with Iodophore-based pre-milking udder preparation while, the third group was treated with Quaternary Ammonium Compound-based pre-milking udder preparation. Raw milk samples were collected from individual dairy animals (20 samples) and bulk tank milk (20 samples) after application of the sanitizers for two successive weeks. Each sample was evaluated for prevalence Aerobic mesophilic count, Coliforms, Staphylococci and Enterococci counts. Results: By the end of the second week of application of sanitizers, the count of aerobic mesophilic count, Coliforms, enterococci and staphylococci counts showed reduction% by 30.22, 36.9, 53.17 and 94.38% for Iodophore-treated group while the Quaternary Ammonium Compound treated group showed reduction% by 22.12, 10.70, 17.71 and 85.41%, respectively in comparing with control group. Conclusion: Our study indicated that Iodophore was better than QACs in reducing the microbial counts of tested milk samples as Iodophore efficacy has a long-term residual disinfection including bacterial re-growth and biofilm control.

Efectividad del sistema rotatorio Mtwo® e hipoclorito de sodio en la eliminación del Enterococcus faecalis. Revisión de la literatura

El objetivo de este estudio fue determinar la efectividad del sistema rotatorio Mtwo® y del hipoclorito de sodio en la eliminación de Enterococcus faecalis.Se realizó una revisión de la literatura de estudios experimentales In vitro y Ex vivo, publicados desde 2010 hasta 2020 en las bases de datos MEDLINE/PubMed, EBSCOhost y Scopus, disponibles en texto completo, publicados en español e inglés. Se seleccionaron los artículos de acuerdo con los criterios establecidos y se analizó el contenido de cada uno de ellos por medio de una tabla matriz en la cual se incluyeron los principales hallazgos.Se identificaron cuarenta y cuatro artículos, de los cuales se incluyeron cinco estudios. La efectividad de la solución irrigadora (NaOCl) demostró resultados a concentraciones de 2.5% y 5.25% y la aplicación del sistema rotatorio Mtwo® reportó disminución del recuento bacteriano al compararlo con otros métodos de instrumentación, así como un recrecimiento menor de bacterias después del tratamiento con este sistema.Las concentraciones de hipoclorito de sodio al 2,5% y al 5,25% y el uso del sistema de instrumentación Mtwo® son eficaces en la eliminación del Enterococcus faecalis.

A Pilot Study Combining Ultrafiltration with Ozonation for the Treatment of Secondary Urban Wastewater: Organic Micropollutants, Microbial Load and Biological Effects

Ozonation followed by ultrafiltration (O3 + UF) was employed at pilot scale for the treatment of secondary urban wastewater, envisaging its safe reuse for crop irrigation. Chemical contaminants of emerging concern (CECs) and priority substances (PSs), microbial load, estrogenic activity, cell viability and cellular metabolic activity were measured before and immediately after O3 + UF treatment. The microbial load was also evaluated after one-week storage of the treated water to assess potential bacteria regrowth. Among the organic micropollutants detected, only citalopram and isoproturon were not removed below the limit of quantification. The treatment was also effective in the reduction in the bacterial loads considering current legislation in water quality for irrigation (i.e., in terms of enterobacteria and nematode eggs). However, after seven days of storage, total heterotrophs regrew to levels close to the initial, with the concomitant increase in the genes 16S rRNA and intI1. The assessment of biological effects revealed similar water quality before and after treatment, meaning that O3 + UF did not produce detectable toxic by-products. Thus, the findings of this study indicate that the wastewater treated with this technology comply with the water quality standards for irrigation, even when stored up to one week, although improvements must be made to minimise microbial overgrowth.

Green photocatalytic disinfection of real sewage: efficiency evaluation and toxicity assessment of eco-friendly TiO2-based magnetic photocatalyst under solar light

To evaluate the green photocatalytic disinfection for practical applications, disinfection of different types of real sewage using magnetic photocatalyst RGO/Fe,N-TiO2/Fe3O4@SiO2 (RGOFeNTFS) under simulated solar light was investigated: low-salinity sewage after tertiary treatment, low-salinity sewage after secondary biological treatment, high-salinity sewage after secondary biological treatment, and high-salinity sewage after chemically enhanced primary treatment. The classification of the sewage as high and low-salinity is based on the regions of sewage source that use seawater and freshwater for toilet flushing, respectively. It shows potential of solar-light-driven photocatalytic disinfection in low-salinity sewage: around 20 min (for sewage after tertiary treatment) and 45 min (for sewage after secondary treatment) of photocatalytic disinfection are required for sewage to meet the discharge standard, and no bacterial regrowth is observed in the treated sewage after 48 h. However, due to the poorer water quality, the high-salinity sewage requires a relatively long reaction time (more than 240 min) to meet the discharge standard, showing minimal practical significance. Further, the complex characteristics of real sewage, such as organic matter, suspended matter, multivalent-ions, pH and DO level significantly influence photocatalytic disinfection, and should be carefully reviewed in evaluating the photocatalytic disinfection of sewage. Besides, RGOFeNTFS shows a good reusability over three cycles for photocatalytic disinfection of low-salinity sewage samples. Moreover, the non-toxicity, indicated by phytoplankton in seawater, of both RGOFeNTFS (<= 3 g/L) and treated low-salinity sewage demonstrates the feasibility of the practical application of photocatalytic disinfection using RGOFeNTFS under irradiation of solar light.

Characterization of Vibrio alginolyticus bacteriophage recovered from shrimp ponds in south west of Iran

Farming of the pacific white leg shrimp (Penaeus vannamei), consists the greatest percent of aquaculture production of crustaceans all over the world. Vibriosis caused by Vibrio species such as Vibrio alginolyticus, is a serious limiting agent for aquaculture. Phage therapy is an eco-friendly alternative to antibiotics as a prophylactic measure for the growth of pathogenic bacteria. The aim of this study was isolation and characterization of Vibriophages present in Litopenaeus vannamei breeding and culture facilities and estuarine resources in south west of Iran, Choebdeh shrimp farming site of Abadan city, Khuzestan Province, based on morphological structure, biological conditions and their potential applications in lysis of Vibrio bacteria. The bacterial and phage samples were analyzed using different physicochemical parameters, 16S rRNA gene sequencing and TEM. The results showed the V. alginolyticus isolates and identified the Podoviridae family of phages. These bacterial isolates were resistant to ampicillin and penicillin. The efficacy of the Podoviridae phage against V. alginolyticus bacterium indicated that this phage has a short period of 30 minutes latent phase, possessed a desirable stability and growth in keeping for 1 hour at 30-55oC, the salinity of 0.5-10 and pH of 5-8. Based on evaluation of phage therapy, mean absorbance values (OD600), in control and treatment groups, at 12 and 24 hours after phage inoculation, were significantly different (p <0.05). According to the results, phage therapy with single-phage suspensions did not prevent bacterial regrowth after treatment. However, delay in development of resistance is a desirable success.

Are iron chelates suitable to perform photo-Fenton at neutral pH for secondary effluent treatment?

There have been concerns about which iron chelate is most suitable for application in the photo-Fenton process as well as the fate of these chelates after application. In this study, five chelating agents, i.e. citric acid (CA), oxalic acid (OA), ethylenediamine disuccinic acid (EDDS), ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA), representing the most used iron chelates were assessed for suitability of application in homogeneous photo-Fenton-like process at pH of 7. The efficiency of the iron chelates were assessed in the disinfection of secondary effluent. The results for the disinfection and bacteria regrowth followed the order EDTA>OA>NTA>CA>OA. All the iron chelates were observed to have increased the COD of the effluent with EDDS having the highest COD contribution. The ability of the chelates to remove aromaticity was measured by the UV254 measurement. The efficiency of the chelates to remove aromaticity decreased in the order CA>EDDS>NTA>CA>OA. To determine the fate of the chelates, toxicity tests were conducted on the chelates before and after irradiation and the results revealed a decrease in toxicity after photoirradiation, implying the chelates were degraded and the products/intermediates produced were of less toxicity as compared to the parent compounds.

Activities of Cefiderocol with Simulated Human Plasma Concentrations against Carbapenem-Resistant Gram-Negative Bacilli in an In Vitro Chemostat Model.

Activities of cefiderocol under simulated human plasma concentrations at the recommended dosing regimen of 2 g every 8 h with a 3-h infusion were evaluated using an in vitro chemostat model. Against a total of 6 meropenem-resistant Gram-negative strains with cefiderocol MICs of 0.5 to 4 μg/ml, including metallo-β-lactamase producers and carbapenem-resistant Acinetobacter baumannii, cefiderocol treatment showed a bactericidal effect within 8 h and sustained efficacy with no marked bacterial regrowth over 24 h.

Optimizing pharmacokinetics/pharmacodynamics of β-lactam/β-lactamase inhibitor combinations against high inocula of ESBL-producing bacteria

Reduced in vitro β-lactam activity against a dense bacterial population is well recognized. It is commonly attributed to the presence of β-lactamase(s) and it is unknown whether the inoculum effect could be diminished by a β-lactamase inhibitor. We evaluated different β-lactam/β-lactamase inhibitor combinations in suppressing a high inoculum of ESBL-producing bacteria. Three clinical isolates expressing representative ESBLs (CTX-M-15 and SHV-12) were examined. The impact of escalating β-lactamase inhibitor (tazobactam or avibactam) concentrations on β-lactam (piperacillin or ceftazidime) MIC reduction was characterized by an inhibitory sigmoid Emax model. The effect of various dosing regimens of β-lactam/β-lactamase inhibitor combinations was predicted using %T>MICi and selected exposures were experimentally validated in a hollow-fibre infection model over 120 h. The threshold exposure to suppress bacterial regrowth was identified using recursive partitioning. A concentration-dependent reduction in β-lactam MIC was observed (r2 ≥0.93). Regrowth could be suppressed in all six experiments using %T>MICi ≥73.6%, but only one out of six experiments below the threshold (P = 0.015). The exposures to suppress regrowth might be attained using the clinical dose of avibactam, but a much higher dose than the standard dose would be needed for tazobactam. A dense population of ESBL-producing bacteria could be suppressed by an optimized dosing regimen of selected β-lactam/β-lactamase inhibitor combinations. The reversibility of enzyme inhibition could play an important role in diminishing the inoculum effect. In vivo investigations to validate these findings are warranted.

Properties of a modified quaternary ammonium silane formulation as a potential root canal irrigant in endodontics

ObjectivesEvaluate a new modified quaternary ammonium silane irrigant solution for its antimicrobial, cytotoxic and mechanical properties of dentine substrate. MethodsRoot canal preparation was performed using stainless steel K-files™ and F4 size protaper with irrigation protocols of 6% NaOCl + 2% CHX; 3.5% QIS; 2% QIS and sterile saline. Biofilms were prepared using E. faecalis adjusted and allowed to grow for 3 days, treated with irrigants, and allowed to grow for 7 days. AFM was performed and surface free energy calculated. MC3T3 cells were infected with endo irrigant treated E. faecalis biofilms. Raman spectroscopy of biofilms were performed after bacterial re-growth on root dentine and exposed to different irrigation protocols and collagen fibers analysed collagen fibers using TEM. Antimicrobial potency against E. faecalis biofilms and cytoxicity against 3T3 NIH cells were also. Resin penetration and MitoTracker green were also evaluated for sealer penetration and mitochondrial viability. Data were analysed using One-way ANOVA, principal component analysis and post-hoc Fisher’s least-significant difference. ResultsElastic moduli were maintained amongst control (5.5 ± 0.9) and 3.5% QIS (4.4 ± 1.1) specimens with surface free energy higher in QIS specimens. MC3T3 cells showed reduced viability in 6%NaOCl+2%CHX specimens compared to QIS specimens. DNA/purine were expressed in increased intensities in control and 6% NaOCl + 2% CHX specimens with bands around 480−490 cm−1 reduced in QIS specimens. 3.5% QIS specimens showed intact collagen fibrillar network and predominantly dead bacterial cells in confocal microscopy. 3.5% QIS irrigant formed a thin crust-type surface layer with cytoplasmic extensions of 3T3NIH spread over root dentine. Experiments confirmed MitoTracker accumulation in 3.5% treated cells. SignificanceNovel QIS root canal irrigant achieved optimum antimicrobial protection inside the root canals facilitating a toxic effect against the Enterococcus faecalis biofilm. Root dentine substrates exhibited optimum mechanical properties and there was viability of fibroblastic mitochondria.