Bacterial Regrowth Research Articles

Production of biostable drinking water using a lab-scale biological trickling filter enriched with hydrogen-oxidizing bacteria

Safeguarding the drinking water quality remains a challenge from the production site to the tap. Alternatively to chemical disinfection, biostable drinking water could serve as a more sustainable approach to produce microbially safe drinking water and to maintain the microbial quality in the drinking water distribution system (DWDS). In this study, the potential of hydrogen-oxidizing bacteria (HOB) to produce biostable drinking water was examined in a continuous trickling filter supplied with hydrogen gas. A biofilm was naturally enriched for 5 months and the bacterial regrowth, invasion potential, and nutrient composition of the water were determined. Treatment improved the biostability significantly, and it is hypothesized that nutrient limitation, especially phosphorous, was a driving force. As a result, the regrowth and invasion potential were lowered, as shown with specific biostability bioassays. Overall, this study demonstrates the proof-of-concept of HOB for producing biostable drinking water through nutrient limitation.

Enhanced removal of biolabile oxygen-depleted dissolved organic matter by coagulation-adsorption process Improves biological stability of reclaimed water

Ensuring biological stability, signifying the maintenance of an unchanged bacterial concentration and composition during water distribution, is essential for mitigating the microbial hazards of reclaimed water. However, the interplay between chlorine-resistant bacteria (CRB) regrowth and molecular transformation of biodegradable organic matter in chlorinated reclaimed water distribution system remain unclear. This work investigated the transformation of dissolved organic matter (DOM) in reclaimed water treated by coagulation and by coagulation-adsorption over a 20-day incubation following chlorination, as well as its interactions with CRB through high-resolution mass spectrometer and high-throughput sequencing. The DOM biotransformation profile and DOM-bacteria interaction network collectively revealed the overall substrate preference and metabolic pattern of CRB (e.g., Methylobacterium-Methylorubrum, Acidovorax, and Sphingomonas), which evinced a propensity for utilizing oxygen-depleted DOM (O/C < 0.5) and producing oxygen-enriched DOM (O/C > 0.4). The incorporation of powdered activated carbon during coagulation markedly decreased the level of biodegradable DOM (4.33 mg C/L) and subsequently retarded the regrowth of CRB by one day compared to coagulation alone, attributable to the selective adsorption of DOM molecules with low O/C onto the activated carbon. This work underscores the critical role of the enhanced removal of oxygen-depleted DOM in ameliorating the biological stability of waters.

Efficient control of antibiotic resistance in wastewater by UV/peracetic acid treatment: Unveiling distinct mechanisms behind the elimination of various contaminants

The abuse of antibiotics has led to an increase in contamination by antibiotics, antibiotic-resistant bacteria (ARB), and antibiotic resistance genes (ARGs) in the environment, especially in wastewater treatment plants, which are the main treatment sites for antibiotic contaminants. In this study, we demonstrated that the combined ultraviolet (UV)/peracetic acid (PAA) process effectively removes coexisting antibiotics, ARB, and ARGs simultaneously by removing 96 % of ampicillin (AMP, with an initial concentration of 10 μM) in 8 min and completely inactivating super-resistant Escherichia coli (E. coli) NDM-1 (i.e., achieving 7.44-log reduction) in just 3 min (PBS, pH=7.0). These bacteria did not proliferate during the 86-h bacterial regrowth test. The analysis of radical quenching and electron paramagnetic resonance revealed that reactive species, such as hydroxyl radicals (·OH) and carbon-centered organic radicals (R-C·), played dominant roles in AMP removal, whereas UV irradiation was crucial for ARB inactivation. Furthermore, the ARGs fragments were efficiently damaged, as demonstrated by gel electrophoresis analysis, leaving few intact sequences. This strategy effectively reduced the horizontal transfer frequency of ARGs. Subsequent evaluations of the stability and efficiency of UV/PAA for pollutant removal in real wastewater further affirmed the broad applicability of the UV/PAA process. This study revealed that UV/PAA system has the potential to alleviate the burden of antibiotic resistance and contribute to a better comprehension and application for novel technology to control antibiotic resistance spread.

Prospectively predicting BPaMZ phase IIb/III trial outcomes using a translational mouse-to-human platform.

Despite known treatments, tuberculosis (TB) remains the world's top infectious killer, highlighting the pressing need for new drug regimens. To prioritize the most efficacious drugs for clinical testing, we previously developed a PK-PD translational platform with bacterial dynamics that reliably predicted short-term monotherapy outcomes in Phase IIa trials from preclinical mouse studies. In this study, we extended our platform to include PK-PD models that account for drug-drug interactions in combination regimens and bacterial regrowth in our bacterial dynamics model to predict cure at the end of treatment and relapse 6 months post-treatment. The Phase III STAND trial testing a new regimen comprised of pretomanid (Pa), moxifloxacin (M), and pyrazinamide (Z) (PaMZ) was suspended after a separate ongoing trial (NC-005) suggested that adding bedaquiline (B) to the PaMZ regimen would improve efficacy. To forecast if the addition of B would, indeed, benefit the PaMZ regimen, we applied an extended translational platform to both regimens. We predicted currently available short- and long-term clinical data well for drug combinations related to BPaMZ. We predicted the addition of B to PaMZ to shorten treatment duration by 2 months and to have similar bacteriological success to standard HRZE treatment (considering only treatment success but not withdrawal from side effects and other adverse events), both at the end of treatment for treatment efficacy and 6 months after treatment has ended in relapse prevention. Using BPaMZ as a case study, we have demonstrated our translational platform can predict Phase II and III outcomes prior to actual trials, allowing us to better prioritize the regimens most likely to succeed.

Pre-exposure to peracetic acid followed by UV treatment for deactivating vancomycin-resistant Enterococcus faecalis through intracellular attack

Antimicrobial resistance (AMR) poses a global health threat to aquatic environments and its propagation is a hot topic. Therefore, deactivating antibiotic-resistant bacteria (ARB) and removing antibiotic resistance genes (ARGs) from water is crucial for controlling AMR transmission. Peracetic acid (PAA), which is known for its potent oxidizing properties and limited by-product formation, is emerging as a favorable disinfectant for water treatment. In this study, we aimed to assess the efficacy of pre-exposure to PAA followed by UV treatment (PAA-UV/PAA) compared with the simultaneous application of UV and PAA (UV/PAA). The focus was on deactivating vancomycin-resistant Enterococcus faecalis (VREfs), a typical ARB in water. Pre-exposure to PAA significantly enhanced the efficacy of subsequent UV/PAA treatment. At a UV fluence of 7.2 mJ cm−2, the PAA-UV/PAA method achieved a 6.21 log reduction in VREfs, surpassing the 1.29 log reduction observed with UV/PAA. Moreover, compared to UV/PAA, PAA-UV/PAA showed increased efficacy with longer pre-exposure times and higher PAA concentrations, maintaining superior performance across a broad pH range and in the presence of humic acid. Flow cytometry analysis indicated minimal cellular membrane damage using both methods. However, the assessments of superoxide dismutase (SOD) activity and adenosine triphosphate content revealed that PAA-UV/PAA induced greater oxidative stress under similar UV irradiation conditions, leading to slower bacterial regrowth. Specifically, SOD activity in PAA-UV/PAA surged to 3.06 times its baseline, exceeding the 1.73-fold increase under UV/PAA conditions. Additionally, pre-exposure to PAA amplified ARGs degradation and reduced resistance gene leakage, effectively mitigating the spread of AMR. Pre-exposure to 200 μM PAA for 10 and 20 min enhanced vanB gene removal efficiency by 0.14 log and 1.29 log, respectively. Our study provides a feasible approach for optimizing UV/PAA disinfection for efficient removal of ARB and ARGs.

Potential of bacteriophage phT4A as a biocontrol agent against Escherichia coli in food matrices

Escherichia coli is one of the most prevalent foodborne pathogens, frequently found in meat and dairy products. Current decontamination methods are often associated with changes in organoleptic characteristics, nutrient loss, and potentially harmful side effects. Furthermore, despite the array of available methods, foodborne outbreaks still frequently occur. For this reason, bacteriophages (or simply phages) emerged as a natural alternative for the biocontrol of bacterial contamination in food without altering their organoleptic properties. In this study, the potential of phage phT4A was assessed in the biocontrol of E. coli in liquid (milk) and solid (ham) food matrices. Firstly, as foods have different pH and temperature values, the influence of these parameters on phage phT4A viability was also assessed to develop an effective protocol. Phage phT4A proved to be stable for long storage periods at pH 7–8 (56 days) and temperatures of 4–37 °C (21 days). Before application of phages to inactivate pathogenic bacteria in food, previous assays were carried out in Tryptic Soy Broth (TSB) to study the dynamics of phage-bacteria interaction. Then, the antibacterial potential of phage phT4A was evaluated in the two food matrices at different temperatures (4, 10 and 25 °C). This phage was more efficient at 25 °C in all tested matrices (maximum inactivation of 6.6, 3.9 and 1.8 log CFU/mL in TSB, milk and ham, respectively) than at 10 °C (maximum decrease of 4.7, 2.1 and 1.0 log CFU/mL in TSB, milk and ham, respectively) and 4 °C (maximum reduction of 2.6 and 0.7 log CFU/mL in TSB and milk, respectively). However, the decrease of temperature from 25 °C to 10 and 4 °C prevented bacterial regrowth. The results suggest that during phage treatment, a balance between an incubation temperature that provide effective results in terms of bacterial inactivation by the phages and at the same time prevents or delays bacterial regrowth, is needed. The application of phage phT4A at a temperature of 10 °C can be an effective strategy in terms of bacterial inactivation, delaying bacterial regrowth and also reducing energy costs.

Survival and Growth of Asellus aquaticus on Different Food Sources from Drinking Water Distribution Systems

Invertebrates, including Asellidae, are part of the natural ecosystem of the drinking water distribution system (DWDS) and are known to cause a nuisance to consumers. In addition, recently, the potential role of the species Asellus aquaticus (L. 1758) in the regrowth of Aeromonas bacteria was published. Aeromonas is included in the Dutch drinking water guidelines as a process parameter, and the guideline values are regularly exceeded. Although neither A. aquaticus nor Aeromonas is associated with health risks, the Evides drinking water utility shows a strong interest in the possible reasons for these exceedances and possible control measures. In surface waters, Asellidae feed mainly on decaying leaves that are abundantly present. These food sources are not present in the DWDS. Therefore, we determined suitable food sources for A. aquaticus in the DWDS. Laboratory experiments show that A. aquaticus individuals survive on biofilm on pipe wall material and loose deposits (sediments) collected from DWDS. Growth and survival rates on these loose deposits were even higher than on the positive control (decaying leaves). As the basis of these loose deposits is inorganic (iron deposits, sand, and pipe particles), the organic matter (living and decaying bacteria, protozoans, fungi, and invertebrates) must be their substrate. These experiments validate hypotheses that Asellidae can grow and survive on organic matter in deposits in DWDS.

Electrochemical removal of antibiotics and multi-drug resistant bacteria using S-functionalized graphene sponge electrodes

In this study, we synthesized S-functionalized graphene sponge electrode and applied it for electrochemical oxidation of five commonly used antibiotics, namely sulfamethoxazole, trimethoprim, ofloxacin, roxithromycin and erythromycin, and the inactivation of a multi-drug resistant Escherichia coli (E. coli). The experiments were performed using real drinking water in a flow-through, one-pass mode. Highly polar antibiotics such as sulfamethoxazole did not adsorb onto the graphene sponge but were completely removed (i.e., ≥95% removal) at low applied current densities (14.5 A m−2). Antibiotics with high affinity for π-π interactions such as ofloxacin were completely removed already in the open circuit, and current application led to their further degradation. S-doped graphene sponge anode resulted in 4.5 log removal of a multi-drug resistant E. coli at 29 A m−2. There was no regrowth of bacteria observed during storage of the electrochemically treated samples, suggesting that the treatment severely impacted the cell viability. Further E. coli removal of 0.7 log was observed after the storage of electrochemically treated samples. The energy consumption of a continuously operated electrochemical system that achieved 4.5 log inactivation of a multi-drug resistant E. coli and 87–99% removal of antibiotics was 1.1 kWh m−3.

Examining how media age affects organic matter removal in activated carbon filtration

AbstractWater samples from Waiora Drinking Water Treatment Plant in New Zealand were analyzed using excitation‐emission matrix fluorescence spectroscopy (EEMS) and parallel factor (PARAFAC) analysis to evaluate organic matter removal across the plant. The assessment also included the individual granular activated carbon (GAC) filters since the filters had varying media ages due to partial media replacement over a 10‐month period, presenting a unique assessment opportunity. PARAFAC analysis identified humic‐like, tyrosine‐protein‐like, and tryptophan‐protein‐like components representing fluorescent dissolved organic matter groups. The humic‐like component strongly correlated with total organic carbon (TOC) concentration and removal was significantly influenced by filter media age. However, protein‐like components had minimal TOC correlation and were not effectively removed by the overall plant treatment irrespective of filter media age. These findings have implications for disinfection, taste and odor, and bacterial regrowth and require an improved media replacement strategy. Further study of the protein‐like components is required.

A flow-through Ti4O7 membrane electrode for ballast water treatment: Performance, mechanism, and comparison with Ti/RuO2-IrO2 electrode

Electrochemical Advanced Oxidation Processes (EAOPs) present a promising solution for ballast water treatment. However, due to the extremely short half-life of reactive oxygen species, the disinfection active zone is limited to the surface of the electrode, resulting in low disinfection current efficiency of EAOPs. To address this challenge, this study developed a flow-through electrochemical disinfection system utilizing a Ti4O7 membrane electrode. A comparative analysis was conducted against a conventional Ti/RuO2-IrO2 electrode to assess the Ti4O7 membrane electrode’s efficacy in ballast water disinfection. The flow-through mode significantly increased the probability of contact between E. coli and the surface of Ti4O7 membrane electrode, consequently leading to a notable increase in the inactivation rate from 1.05 min−1 to 1.63 min−1, at a current density of 20 mA/cm2, at a Cl- concentration of 16.5 g/L, and an initial E. coli concentration of 107 CFU/mL. Notably, at low current densities, the Ti4O7 membrane electrode exhibited superior disinfection performance compared to the Ti/RuO2-IrO2 electrode. The predominant disinfection active species in the Ti4O7 membrane electrode system were identified as reactive oxygen species, particularly •OH and •O2–, contrasting with the abundant generation of active chlorine species (ACSs) observed in the Ti/RuO2-IrO2 electrode system. Both electrode systems demonstrated the ability to disrupt the cellular structure of E. coli. However, due to the sustained effectiveness of ACSs, the degradation rate of total protein in the Ti/RuO2-IrO2 electrode system (40.53 %) surpassed that of the Ti4O7 membrane electrode system (12.39 %). Despite the minimal concentration of ACSs in the Ti4O7 membrane system (0.02 mg/L), no bacterial regeneration was observed, highlighting the efficacy of the Ti4O7 membrane electrode in preventing bacterial regrowth. This study elucidated a free radical-based disinfection mechanism for the Ti4O7 membrane electrode, affirming its feasibility for ballast water disinfection purposes.

Occurrence and Mitigation of Bacterial Regrowth in Stored Household Water in Eastern Coastal Madagascar

In communities where people lack on-demand, safely managed drinking water, stored household water often becomes contaminated by fecal bacteria, regardless of the source-water quality. The objectives of this paper are to assess and control bacterial contamination in stored household water in Toamasina, a rapidly urbanizing city in eastern coastal Madagascar. We collected samples of source water and stored household water from 10 representative households that use different water sources and different storage strategies, and we analyzed the samples for several fecal indicator bacteria. We also tested three methods that residents of Toamasina could realistically employ for cleaning their household water storage vessels, assessing the effect of the cleaning methods on measured bacterial levels in the water. Consistent with the previous literature, we found that concentrations of total coliforms in stored household water were significantly higher than in the corresponding source water (p &lt; 0.05). In 100% of households that stored their water in 20 L polyethylene jerrycans (n = 4), biofilms on the walls of the jerrycan harbored total coliforms and Enterococcus. The use of a closed storage container was, on its own, not found to provide a meaningful protective effect against bacterial regrowth; to be protective, closed storage containers must be combined with high-quality source water and/or with adequate cleaning to prevent biofilm formation. A dilute solution of sodium hypochlorite, known locally as Sûr’Eau or Manadio Rano, was both the most effective and the least expensive method for cleaning household water storage containers. We conclude that regular and effective cleaning of storage containers is an essential component of safe water storage. Because household storage of collected water is common in many low- and middle-income countries, these results are important towards the worldwide achievement of the United Nations’ Sustainable Development Goal 6.

Centroid of the bacterial growth curves: a metric to assess phage efficiency

Phage replication can be studied using various approaches, including measuring the optical density (OD) of a bacterial culture in a liquid medium in the presence of phages. A few quantitative methods are available to measure and compare the efficiency of phages by using a single index based on the analysis of OD curves. However, these methods are not always applicable to non-canonical OD curves. Using the concept of center of area (centroid), we developed a metric called Centroid Index (CI), sensitive to the trend of the growth curves (OD distribution) including bacterial regrowth, which is not considered by the methods already available. We also provide a user-friendly software to facilitate the calculation of CI. This method offers an alternative and more precise way to determine phage efficiency by considering the OD variations over time, which may help in the selection of phages for biocontrol applications.

Impact of nutritional factors on in vitro PK/PD modelling of polymyxin B against various strains of Acinetobacter baumannii

The main objective of this study was to assess the effect of rich artificial cation adjusted Mueller-Hinton broth (CAMHB), on the growth of three Acinetobacter baumannii strains, ATCC 19606 and two clinical A. baumannii strains, either susceptible or resistant to polymyxin B (PMB), and on the PMB bactericidal activity. A pharmacokinetic (PK) / pharmacodynamic (PD) modelling approach was then used to characterize the effect of PMB in various conditions.Time-kill experiments were performed using undiluted CAMHB or diluted at 50%, 25% and 10%, with or without Ca2+ and Mg2+ compensation (known to affect PMB activity), and with PMB concentrations ranging from 0.25 mg/L to 256 mg/L based on the strain's MIC. For each strain, time-kill replicates were modelled using NONMEM®.Unexpectedly, CAMHB dilution by up to 10-fold did not affect the growth rate of any of the three strains in the absence of PMB. Yet, PMB bactericidal activity increased with medium dilution resulting in particular in a reduction of the apparent bacterial regrowth of the various strains observed typically after few hours of experiment. The data of each strain were well characterised by a PK/PD model with two bacterial sub-populations with different susceptibility to PMB (a more susceptible (S+) and a less susceptible (S−)). Relatively large, unexplained strain-to-strain variability was observed regarding the impact of medium dilution as well as cation compensation. Complementary experiments are needed to characterise the mechanism underlying this medium dilution effect.

Constituents of Coliform Species Contained in the Permeate of Microfiltration Membranes in Wastewater Treatment

MBRs (Membrane bioreactors) have been increasingly employed for municipal and industrial wastewater treatment in the last decades for their small footprint and excellent effluent quality. However, microorganisms are often detected in the permeates of microfiltration (MF) membranes even with small pore sizes. Coliform bacteria are known for indicating the potential presence of pathogenic bacteria that cause infectious disease such as bacteremia, respiratory tract infections, and urinary tract infections. Thus, the retention of coliform bacteria by membrane processes is important when the membrane process is utilized in water reclamation. In this study, a microbial community of coliform bacteria in the permeates of MF membranes with different pore sizes (0.2, 0.4, and 0.8 µm) was identified. The results showed that the dominant coliform bacteria changed from Enterobacter spp. and Citrobacter spp. in the activated sludge to Enterobacter spp. and Klebsiella spp. in the permeate of MF membranes, while some pieces of membranes showed complete retention. The bacterial regrowth on the surface of the piping system on the permeate side could be a significant factor contributing to the frequent and exclusive detection of Enterobacter spp. and Klebsiella spp. in the case of membranes with small pore size (0.2 and 0.4 µm) after a long continuous filtration time. To indicate the public health-related risk of treated wastewater by MF, Escherichia coli may not be a suitable indicator species because E. coli is relatively retentive in MF compared to other coliforms.

Characterization and therapeutic potential of MRABP9, a novel lytic bacteriophage infecting multidrug-resistant Acinetobacter baumannii clinical strains

Acinetobacter baumannii is one of the most important pathogens of healthcare-associated infections. The rising prevalence of multidrug-resistant A. baumannii (MRAB) strains and biofilm formation impact the outcome of conventional treatment. Phage-related therapy is a promising strategy to tame troublesome multidrug-resistant bacteria. Here, we isolated and evaluated a highly efficient lytic phage called MRABP9 from hospital sewage. The phage was a novel species within the genus Friunavirus and exhibited lytic activity against 2 other identified MRAB strains. Genomic analysis revealed it was a safe virulent phage and a pectate lyase domain was identified within its tail spike protein. MRABP9 showed potent bactericidal and anti-biofilm activity against MRAB, significantly delaying the time point of bacterial regrowth in vitro. Phage administration could rescue the mice from acute lethal MRAB infection. Considering its features, MRABP9 has the potential as an efficient candidate for prophylactic and therapeutic use against acute infections caused by MRAB strains.

Synergistic effects of inhaled aztreonam plus tobramycin on hypermutable cystic fibrosis Pseudomonas aeruginosa isolates in a dynamic biofilm model evaluated by mechanism-based modelling and whole genome sequencing

ObjectiveHypermutable Pseudomonas aeruginosa strains are highly prevalent in chronic lung infections of patients with cystic fibrosis (CF). Acute exacerbations of these infections have limited treatment options. This study aimed to investigate inhaled aztreonam and tobramycin against clinical hypermutable P. aeruginosa strains using the CDC dynamic in vitro biofilm reactor (CBR), mechanism-based mathematical modelling (MBM) and genomic studies. MethodsTwo CF multidrug-resistant strains were investigated in a 168 h CBR (n = 2 biological replicates). Regimens were inhaled aztreonam (75 mg 8-hourly) and tobramycin (300 mg 12-hourly) in monotherapies and combination. The simulated pharmacokinetic profiles of aztreonam and tobramycin (t1/2 = 3 h) were based on published lung fluid concentrations in patients with CF. Total viable and resistant counts were determined for planktonic and biofilm bacteria. MBM of total and resistant bacterial counts and whole genome sequencing were completed. ResultsBoth isolates showed reproducible bacterial regrowth and resistance amplification for the monotherapies by 168 h. The combination performed synergistically, with minimal resistant subpopulations compared to the respective monotherapies at 168 h. Mechanistic synergy appropriately described the antibacterial effects of the combination regimen in the MBM. Genomic analysis of colonies recovered from monotherapy regimens indicated noncanonical resistance mechanisms were likely responsible for treatment failure. ConclusionThe combination of aztreonam and tobramycin was required to suppress the regrowth and resistance of planktonic and biofilm bacteria in all biological replicates of both hypermutable multidrug-resistant P. aeruginosa CF isolates. The developed MBM could be utilised for future investigations of this promising inhaled combination.

Effect of dissolved organic matter on bacterial regrowth and response after ultraviolet disinfection

The effect of dissolved organic matter (DOM) on bacterial regrowth in water after disinfection using ultraviolet (UV) light emitting diodes (UVLEDs) is still unclear. Herein, the regrowth and responses of Vibrio parahaemolyticus and Bacillus cereus were investigated after being exposed to UVLEDs at combined wavelengths (265 and 280 nm) in a phosphate-buffered saline consisting of Suwannee River natural organic matter (SRNOM) and Suwannee River fulvic acid (SRFA). Low-molecular-weight (MW) organic compounds, which may form into intermediary photoproducts, and indicate bacterial repair metabolism, were characterized through non-target screening using orbitrap mass spectrometry. This study demonstrates the ability of the UVLEDs-inactivated cells to regrow. After UV exposure, a considerable upregulation of RecA was observed in two strains. With increasing the incubation time, the expression levels of RecA in V. parahaemolyticus increased, which may be attributed to the dark repair mechanism. Coexisting anionic DOM affects both the disinfection and bacterial regrowth processes. The time required for bacterial regrowth after UV exposure reflects the time needed for the individual cells to reactivate, and it differs in the presence or absence of DOM. In the presence of DOM, the cells were less damaged and required less time to grow. The UVLEDs exposure results in the occurrence of low-MW organic compounds, including carnitine or acryl-carnitine with N-acetylmuramic acid, which are associated with bacterial repair metabolism. Overall, the results of this study expand the understanding of the effects of water matrices on bacterial health risks. This can aid in the development of more effective strategies for water disinfection.

Model-based learn and confirm: designing effective treatment regimens against multidrug resistant Gram-negative pathogens

Over the last decade, there has been a growing appreciation for the use of in vitro and in vivo infection models to generate robust and informative nonclinical PK/PD data to accelerate the clinical translation of treatment regimens. The objective of this study was to develop a model-based “learn and confirm” approach to help with the design of combination regimens using in vitro infection models to optimise the clinical utility of existing antibiotics. Static concentration time-kill studies were used to evaluate the PD activity of polymyxin B (PMB) and meropenem against two carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates; BAA2146 (PMB-susceptible) and BRKP67 (PMB-resistant). A mechanism-based model (MBM) was developed to quantify the joint activity of PMB and meropenem. In silico simulations were used to predict the time-course of bacterial killing using clinically-relevant PK exposure profiles. The predictive accuracy of the model was further evaluated by validating the model predictions using a one-compartment PK/PD in vitro dynamic infection model (IVDIM). The MBM captured the reduction in bacterial burden and regrowth well in both the BAA2146 and BRKP67 isolate (R2 = 0.900 and 0.940, respectively). The bacterial killing and regrowth predicted by the MBM were consistent with observations in the IVDIM: sustained activity against BAA2146 and complete regrowth of the BRKP67 isolate. Differences observed in PD activity suggest that additional dose optimisation might be beneficial in PMB-resistant isolates. The model-based approach presented here demonstrates the utility of the MBM as a translational tool from static to dynamic in vitro systems to effectively perform model-informed drug optimisation.

Optimizing in vitro phage-ciprofloxacin combination formulation for respiratory therapy of multi-drug resistant Pseudomonas aeruginosa infections

Respiratory infection caused by multi-drug resistant (MDR) Pseudomonas aeruginosa is challenging to treat. In this study, we investigate the optimal dose of anti-pseudomonas phage PEV31 (103, 105, and 108 PFU/mL) combined with ciprofloxacin (ranging from 1/8× MIC to 8× MIC) to treat the MDR P. aeruginosa strain FADD1-PA001 using time-kill studies. We determined the impact of phage growth kinetics in the presence of ciprofloxacin through one-step growth analysis. Single treatments with either phage PEV31 or ciprofloxacin (except at 8× MIC) showed limited bactericidal efficiency, with bacterial regrowth observed at 48 h. The most effective treatments were PEV31 at multiplicity of infection (MOI) of 0.1 and 100 combined with ciprofloxacin at concentrations above 1× MIC, resulting in a >4 log10 reduction in bacterial counts. While the burst size of phage PEV31 was decreased with increasing ciprofloxacin concentration, robust antimicrobial effects were still maintained in the combination treatment. Aerosol samples collected from vibrating mesh nebulization of the combination formulation at phage MOI of 100 with 2× MIC effectively inhibited bacterial density. In summary, our combination treatments eradicated in vitro bacterial growth and sustained antimicrobial effects for 48 h. These results indicated the potential application of nebulization-based strategies for the combination treatment against MDR lung infections.

Effect of flow fluctuation on water pollution in drinking water distribution systems

The detachment of biofilm caused by changes in hydraulic conditions is an essential reason for the pollution of water in the drinking water distribution system (DWDS). In this research, the effect of flow fluctuation on bulk water quality was studied. The turbidity, iron concentration, manganese concentration, the total number of bacteria, biodegradable dissolved organic carbon (BDOC), bacterial community structure, and pathogenic genes in bacteria of bulk water were analyzed. The results indicate that the detachment of biofilm caused by fluctuant flow and reverse flow (especially instant reverse flow) can lead to the pollution of water. Throughout the entire experimental period, the turbidity under fluctuant flow velocity is 4.92%∼49.44% higher than that under other flow velocities. BDOC concentration is 5.68%∼53.99% higher than that under low and high flow velocities. The flow fluctuation increases bacterial regrowth potential (BRP) and reduces the biological stability of the bulk water. Low flow velocity is more conducive to the expression of pathogenic functional genes. In the short term, the water quality under low flow velocity is the best. Nevertheless, in a long-term operation (about seven days later), the water quality under high flow velocity is better than that under other flow velocities. This research brings new knowledge about the fluctuant hydraulic conditions on the bulk water quality within the DWDS and provides data support for stable drinking water distribution.