scholarly journals Classification of In Vitro Phage–Host Population Growth Dynamics

2021 ◽  
Vol 9 (12) ◽  
pp. 2470
Author(s):  
Patricia E. Sørensen ◽  
Duncan Y. K. Ng ◽  
Luc Duchateau ◽  
Hanne Ingmer ◽  
An Garmyn ◽  
...  
Keyword(s):  
Bacterial Growth ◽  
Phage Therapy ◽  
Growth Dynamics ◽  
Host Population ◽  
Growth Patterns ◽  
Bacterial Cells ◽  
Bacterial Killing ◽  
Promising Alternative ◽  
Group 2

The therapeutic use of bacteriophages (phage therapy) represents a promising alternative to antibiotics to control bacterial pathogens. However, the understanding of the phage–bacterium interactions and population dynamics seems essential for successful phage therapy implementation. Here, we investigated the effect of three factors: phage species (18 lytic E. coli-infecting phages); bacterial strain (10 APEC strains); and multiplicity of infection (MOI) (MOI 10, 1, and 0.1) on the bacterial growth dynamics. All factors had a significant effect, but the phage appeared to be the most important. The results showed seven distinct growth patterns. The first pattern corresponded to the normal bacterial growth pattern in the absence of a phage. The second pattern was complete bacterial killing. The remaining patterns were in-between, characterised by delayed growth and/or variable killing of the bacterial cells. In conclusion, this study demonstrates that the phage–host dynamics is an important factor in the capacity of a phage to eliminate bacteria. The classified patterns show that this is an essential factor to consider when developing a phage therapy. This methodology can be used to rapidly screen for novel phage candidates for phage therapy. Accordingly, the most promising candidates were phages found in Group 2, characterised by growth dynamics with high bacterial killing.

scholarly journals Comparative activity of Ceftriaxone, Ciprofloxacin and Gentamicin as a function of bacterial growth rate probed by Escherichia coli chromosome replication in the mouse peritonitis model

2018 ◽  
Author(s):  
Maria Schei Haugan ◽  
Anders Løbner-Olesen ◽  
Niels Frimodt-Møller
Keyword(s):  
Growth Rate ◽  
Bacterial Growth ◽  
Growth Dynamics ◽  
Chromosome Replication ◽  
Bacterial Cells ◽  
Bacterial Growth Rate ◽  
Pre Treatment

AbstractCommonly used antibiotics exert their effect predominantly on rapidly growing bacterial cells, yet growth dynamics taking place during infection in a complex host environment remain largely unknown. Hence, means to measure in situ bacterial growth rate is essential to predict the outcome of antibacterial treatment. We have recently validated chromosome replication as readout for in situ bacterial growth rate during Escherichia coli infection in the mouse peritonitis model. By the use of two complementary methods (qPCR and fluorescence microscopy) for differential genome origin and terminus copy number quantification, we demonstrated the ability to track bacterial growth rate, both on a population average and on a single-cell level; from one single biological specimen. Here, we asked whether the in situ growth rate could predict antibiotic treatment effect during infection in the same model. Parallel in vitro growth experiments were conducted as proof-of-concept. Our data demonstrate that the activity of commonly used antibiotics Ceftriaxone and Gentamicin correlated with pre-treatment bacterial growth rate; both drugs performing better during rapid growth than during slow growth. Conversely, Ciprofloxacin was less sensitive to bacterial growth rate, both in a homogenous in vitro bacterial population and in a more heterogeneous in vivo bacterial population. The method serves as a platform to test any antibiotic’s dependency upon active in situ bacterial growth. Improved insight into this relationship in vivo could ultimately prove helpful in evaluating future antibacterial strategies.ImportanceMost antibiotics in clinical use exert their effect predominantly on rapidly growing bacterial cells, yet there is a lack of insight into bacterial growth dynamics taking place during infection in vivo. We have applied inexpensive and easily accessible methods for extraction of in situ bacterial growth rate from bacterial chromosome replication during experimental murine infection. This approach not only allows for a better understanding of bacterial growth dynamics taking place during the course of infection, but also serves as a platform to test the activity of different antibiotics as a function of pre-treatment in situ growth rate. The method has the advantage that bacterial growth rate can be probed from a single biological sample, with the potential for extension into clinical use in pre-treatment infected biological specimens. A better understanding of commonly used antibiotics’ level of dependency upon bacterial growth, combined with measurements of in situ bacterial growth rate in infected clinical specimens, could prove helpful in evaluating future antibacterial treatment regimens.

scholarly journals Impact of Different Carbon Sources on the in vitro Growth and Viability of Escherichia coli (SUBE01) and Salmonella spp. (SUBS01) Cells

2016 ◽  
pp. 39-44
Author(s):  
Ifra Tun Nur ◽  
Jannatun Tahera ◽  
Md Sakil Munna ◽  
M Majibur Rahman ◽  
Rashed Noor
Keyword(s):  
Escherichia Coli ◽  
Bacterial Growth ◽  
Culture Media ◽  
Bacterial Species ◽  
Carbon Sources ◽  
Growth Dynamics ◽  
Luria Bertani ◽  
Tween 20 ◽  
Salmonella Spp

With a previous observation of Escherichia coli growth cessation along with temperature variation within three different bacteriological culture media (nutrient agar, Luria-Bertani agar and minimal agar), current investigation further depicted on the possible growth dynamics of Escherichia coli (SUBE01) and Salmonella (SUBS01) growth and viability upon supplementation of different carbon sources (dextrose, sucrose, lactose, glycerol and tween 20) at 37°C under the aeration of 100 rpm. Viability of the tested bacterial species was assessed through the enumeration of the colony forming unit (cfu) appeared upon prescribed incubation for 12-24 hours on different agar plates consisting of the above mentioned carbon sources. Besides, to inspect the cellular phenotypic changes, morphological observations were conducted under the light microscope. Variations in bacterial growth (either growth acceleration or cessation) were further noticed through the spot tests on the agar plates. Considerable shortfalls in the culturable cells of E. coli and Salmonella spp. were noted in the minimal media separately consisting of sucrose, lactose, glycerol or tween 20 while an opposite impact of accelerated growth was noticed in the media supplied with dextrose. The data revealed a hierarchy of consequence of carbon sources as nutrient generators whereby the favourable bacterial growth and survival order of the carbon sources was estimated as dextrose > glycerol > lactose > tween 20 > sucrose.Bangladesh J Microbiol, Volume 32, Number 1-2,June-Dec 2015, pp 39-44

scholarly journals Impact of thrombocytes, on bacterial growth and antimicrobial activity of selected antibiotics

2019 ◽  
Vol 39 (3) ◽  
pp. 593-597 ◽  
Author(s):  
Alina Karoline Nussbaumer-Pröll ◽  
Sabine Eberl ◽  
Birgit Reiter ◽  
Thomas Stimpfl ◽  
Walter Jäger ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bacterial Growth ◽  
Limiting Factor ◽  
Growth Media ◽  
Healthy Human ◽  
Bacterial Killing ◽  
Bacteria Growth ◽  
The Impact

AbstractIn vitro pharmacodynamic models are used to optimize in vivo dosing regimens in antimicrobial drug development. One limiting factor of such models is the lack of host factors such as corpuscular blood components as erythrocytes which have already been shown to impact activity of antibiotics and/or growth of the pathogen. However, the impact of thrombocytes has not previously been investigated. We set out to investigate if the addition of thrombocytes (set to physiological concentrations in blood of healthy human, i.e., 5 × 105 thrombocytes/μL standard growth media Mueller Hinton Broth, MHB) has an influence on bacterial growth and on the efficacy of antibiotics against Gram+ and Gram− bacteria. Growth assays and time-killing-curves (TKC) were performed with ATCC-strains of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa in triplicate over 24 h. The same approach was followed for 5 clinical isolates of Escherichia coli. Meropenem, ciprofloxacin, and tigecycline were tested as representatives of broad-spectrum antibiotics, and concentrations several-fold above and below the minimal inhibitory concentration (MIC) were simulated. No significant impact of thrombocytes was found on bacterial growth or antimicrobial stability for the investigated agents. Bacteria reduced thrombocyte content to different degree, indicating direct interaction of pathogens and thrombocytes. Impact on bacterial killing was observed but was not fully reproducible when thrombocytes from different donors where used. While interaction of bacteria and thrombocytes was evident in the present study, interaction between antibiotic activity and thrombocytes seems unlikely. Whether variability was caused by different thrombocyte concentrates needs further investigation.

scholarly journals Extension of Pharmacokinetic/Pharmacodynamic Time-Kill Studies To Include Lipopolysaccharide/Endotoxin Release from Escherichia coli Exposed to Cefuroxime

2020 ◽  
Vol 64 (4) ◽  
Author(s):  
Anders Thorsted ◽  
Eva Tano ◽  
Kia Kaivonen ◽  
Jan Sjölin ◽  
Lena E. Friberg ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bacterial Growth ◽  
Mean Transit Time ◽  
Integrated Analysis ◽  
Antibiotic Exposure ◽  
Bacterial Killing ◽  
Content Type ◽  
Time Courses ◽  
Time Kill

ABSTRACT The release of inflammatory bacterial products, such as lipopolysaccharide (LPS)/endotoxin, may be increased upon the administration of antibiotics. An improved quantitative understanding of endotoxin release and its relation to antibiotic exposure and bacterial growth/killing may be gained by an integrated analysis of these processes. The aim of this work was to establish a mathematical model that relates Escherichia coli growth/killing dynamics at various cefuroxime concentrations to endotoxin release in vitro. Fifty-two time-kill experiments informed bacterial and endotoxin time courses and included both static (0×, 0.5×, 1×, 2×, 10×, and 50× MIC) and dynamic (0×, 15×, and 30× MIC) cefuroxime concentrations. A model for the antibiotic-bacterium interaction was established, and antibiotic-induced bacterial killing followed a sigmoidal Emax relation to the cefuroxime concentration (MIC-specific 50% effective concentration [EC50], maximum antibiotic-induced killing rate [Emax] = 3.26 h−1 and γ = 3.37). Endotoxin release was assessed in relation to the bacterial processes of growth, antibiotic-induced bacterial killing, and natural bacterial death and found to be quantitatively related to bacterial growth (0.000292 endotoxin units [EU]/CFU) and antibiotic-induced bacterial killing (0.00636 EU/CFU). Increased release following the administration of a second cefuroxime dose was described by the formation and subsequent antibiotic-induced killing of filaments (0.295 EU/CFU). Release due to growth was instantaneous, while release due to antibiotic-induced killing was delayed (mean transit time of 7.63 h). To conclude, the in vitro release of endotoxin is related to bacterial growth and antibiotic-induced killing, with higher rates of release upon the killing of formed filaments. Endotoxin release over 24 h is lowest when antibiotic exposure rapidly eradicates bacteria, while increased release is predicted to occur when growth and antibiotic-induced killing occur simultaneously.

scholarly journals Sphingosine kills bacteria by binding to cardiolipin

2020 ◽  
Vol 295 (22) ◽  
pp. 7686-7696 ◽  
Author(s):  
Rabea Verhaegh ◽  
Katrin Anne Becker ◽  
Michael J. Edwards ◽  
Erich Gulbins
Keyword(s):  
Plasma Membrane ◽  
Bactericidal Activity ◽  
Cell Biology ◽  
Plasma Membranes ◽  
Central Element ◽  
Lipid Binding ◽  
Bacterial Cells ◽  
Bacterial Killing

Sphingosine is a long-chain sphingoid base that has been shown to have bactericidal activity against many pathogens, including Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli. We have previously demonstrated that sphingosine is present in nasal, tracheal, and bronchial epithelial cells and constitutes a central element of the defense of the airways against bacterial pathogens. Here, using assorted lipid-binding and cell biology assays, we demonstrate that exposing P. aeruginosa and S. aureus cells to sphingosine results in a very rapid, i.e. within minutes, permeabilization of the bacterial plasma membrane, resulting in leakiness of the bacterial cells, loss of ATP, and loss of bacterial metabolic activity. These alterations rapidly induced bacterial death. Mechanistically, we demonstrate that the presence of the protonated NH2 group in sphingosine, which is an amino-alcohol, is required for sphingosine's bactericidal activity. We also show that the protonated NH2 group of sphingosine binds to the highly negatively–charged lipid cardiolipin in bacterial plasma membranes. Of note, this binding was required for bacterial killing by sphingosine, as revealed by genetic experiments indicating that E. coli or P. aeruginosa strains that lack cardiolipin synthase are resistant to sphingosine, both in vitro and in vivo. We propose that binding of sphingosine to cardiolipin clusters cardiolipin molecules in the plasma membrane of bacteria. This clustering results in the formation of gel-like or even crystal-like structures in the bacterial plasma membrane and thereby promotes rapid permeabilization of the plasma membrane and bacterial cell death.

scholarly journals Modeling of Scale-Dependent Bacterial Growth by Chemical Kinetics Approach

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Haydee Martínez ◽  
Joaquín Sánchez ◽  
José-Manuel Cruz ◽  
Guadalupe Ayala ◽  
Marco Rivera ◽  
...  
Keyword(s):  
Chemical Kinetics ◽  
Bacterial Growth ◽  
Liquid Surface ◽  
Growth Dynamics ◽  
Volume Ratio ◽  
Growth Patterns ◽  
Open Flow ◽  
E Coli ◽  
Bacterial Cultures ◽  
Fitting Procedures

We applied the so-called chemical kinetics approach to complex bacterial growth patterns that were dependent on the liquid-surface-area-to-volume ratio (SA/V) of the bacterial cultures. The kinetic modeling was based on current experimental knowledge in terms of autocatalytic bacterial growth, its inhibition by the metabolite CO2, and the relief of inhibition through the physical escape of the inhibitor. The model quantitatively reproduces kinetic data of SA/V-dependent bacterial growth and can discriminate between differences in the growth dynamics of enteropathogenicE. coli,E. coli  JM83, andSalmonella typhimuriumon one hand andVibrio choleraeon the other hand. Furthermore, the data fitting procedures allowed predictions about the velocities of the involved key processes and the potential behavior in an open-flow bacterial chemostat, revealing an oscillatory approach to the stationary states.

scholarly journals In vitro synergy between sodium deoxycholate and furazolidone against enterobacteria

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Vuong Van Hung Le ◽  
Catrina Olivera ◽  
Julian Spagnuolo ◽  
Ieuan G. Davies ◽  
Jasna Rakonjac
Keyword(s):  
Nitric Oxide ◽  
Growth Inhibition ◽  
Fold Increase ◽  
Sodium Deoxycholate ◽  
Efflux Pumps ◽  
Bacterial Cells ◽  
Bacterial Killing ◽  
E Coli

Abstract Background Antimicrobial combinations have been proven as a promising approach in the confrontation with multi-drug resistant bacterial pathogens. In the present study, we identify and characterize a synergistic interaction of broad-spectrum nitroreductase-activated prodrugs 5-nitrofurans, with a secondary bile salt, sodium deoxycholate (DOC) in growth inhibition and killing of enterobacteria. Results Using checkerboard assay, we show that combination of nitrofuran furazolidone (FZ) and DOC generates a profound synergistic effect on growth inhibition in several enterobacterial species including Escherichia coli, Salmonella enterica, Citrobacter gillenii and Klebsiella pneumoniae. The Fractional Inhibitory Concentration Index (FICI) for DOC-FZ synergy ranges from 0.125 to 0.35 that remains unchanged in an ampicillin-resistant E. coli strain containing a β-lactamase-producing plasmid. Findings from the time-kill assay further highlight the synergy with respect to bacterial killing in E. coli and Salmonella. We further characterize the mechanism of synergy in E. coli K12, showing that disruption of the tolC or acrA genes that encode components of multidrug efflux pumps causes, respectively, a complete or partial loss, of the DOC-FZ synergy. This finding indicates the key role of TolC-associated efflux pumps in the DOC-FZ synergy. Overexpression of nitric oxide-detoxifying enzyme Hmp results in a three-fold increase in FICI for DOC-FZ interaction, suggesting a role of nitric oxide in the synergy. We further demonstrate that DOC-FZ synergy is largely independent of NfsA and NfsB, the two major activation enzymes of the nitrofuran prodrugs. Conclusions This study is to our knowledge the first report of nitrofuran-deoxycholate synergy against Gram-negative bacteria, offering potential applications in antimicrobial therapeutics. The mechanism of DOC-FZ synergy involves FZ-mediated inhibition of TolC-associated efflux pumps that normally remove DOC from bacterial cells. One possible route contributing to that effect is via FZ-mediated nitric oxide production.

Experimental and theoretical studies of tumor growth

2019 ◽  
Vol 04 (03) ◽  
pp. 1950004 ◽  
Author(s):  
Hao Sun ◽  
Timothy Eswothy ◽  
Kerlin P. Robert ◽  
Jiaoyan Li ◽  
L. G. Zhang ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Tumor Growth ◽  
Growth Dynamics ◽  
Cancer Cell Line ◽  
Growth Patterns ◽  
Pressure Fields ◽  
Biological Phenomena ◽  
Growth Mechanics ◽  
Experimental And Theoretical Studies

Most biological phenomena commonly involve growth and expansion mechanics. In this work, we propose an innovative model of cancerous growth which posits that an expandable tumor can be described as a poroelastic medium consisting of solid and fluid components. To verify the feasibility of the model, we utilized an established epithelial human breast cancer cell line (MDA-MB-231) to generate an in vitro tumorsphere system to observe tumor growth patterns in both constrained and unconstrained growth environments. The tumorspheres in both growth environments were grown with and without the FDA-approved anti-breast cancer anthracycline, Doxorubicin (Dox), in order to observe the influence small molecule drugs have on tumor-growth mechanics. In our biologically informed mechanical description of tumor growth dynamics, we derive the governing equations of the tumor’s growth and incorporate them with large deformation to improve the accuracy and efficiency of our simulation. Meanwhile, the dynamic finite element equations (DFE) for coupled displacement field and pressure field are formulated. Moreover, the porosity and growth tensor are generalized to be functions of displacement and pressure fields. We also introduce a specific porosity and growth tensor. In both cases, the formalism of continuum mechanics and DFE are accompanied by accurate numerical simulations.

scholarly journals Phage Targeting Streptococcus mutans In Vitro and In Vivo as a Caries-Preventive Modality

Antibiotics ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1015
Author(s):  
Amit Wolfoviz-Zilberman ◽  
Reut Kraitman ◽  
Ronen Hazan ◽  
Michael Friedman ◽  
Yael Houri-Haddad ◽  
...  
Keyword(s):  
Sustained Release ◽  
Streptococcus Mutans ◽  
Delivery System ◽  
Phage Therapy ◽  
Caries Prevention ◽  
Promising Alternative ◽  
Carious Lesions ◽  
Caries Model

Dental caries is a common infectious disease worldwide. Current conventional therapies lack specific antimicrobial effects against Streptococcus mutans, a key bacterium that induces caries. A promising alternative approach is bacteriophage (phage) therapy. Recently, SMHBZ8 phage targeting S. mutans was isolated and characterized. The aim of this study was to evaluate the caries-prevention efficacy of SMHBZ8 using in vitro and in vivo caries models. Hemi-mandibles dissected from euthanized healthy mice were subjected to caries-promoting conditions in vitro. Jaws treated with phage therapy in suspension and in formulation with a sustained-release delivery system showed no carious lesions, similar to control and chlorhexidine-treated jaws. Subsequently, SMHBZ8 phage suspension also prevented carious lesion development in a murine caries model in vivo. In both models, caries lesions were analyzed clinically and radiographically by µCT scans. This study shows how SMHBZ8 phage therapy targeting S. mutans can serve as an efficient caries-prevention modality, in suspension or with a sustained-release delivery system, by in vitro and in vivo mouse models.

scholarly journals Determination of the optimal bacteriophage dose to control Pseudomonas aeruginosa using evolutionary programming and stochastic kinetics

2016 ◽  
Author(s):  
Diana C Ardila ◽  
Juan D Castro ◽  
Angela V Holguín ◽  
Viviana Clavijo ◽  
Catalina Prada ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Reverse Engineering ◽  
Pathogenic Bacteria ◽  
Phage Therapy ◽  
Evolutionary Programming ◽  
Stochastic Simulations ◽  
Resistant Bacteria ◽  
Multiple Drug ◽  
Promising Alternative

Phage-therapy is a promising alternative against pathogenic, multiple drug resistant bacteria. In this work we propose an algorithm to determine the optimal bacteriophage dose able to minimize a population of Pseudomonas aeruginosa. Reverse engineering was used to determine the kinetic parameters; subsequently, a bi-level optimization platform was implemented for a model based on evolutionary programming. Our prediction of optimal dose was tested in vitro with planktonic cultures of P. aeruginosa. From the data obtained, we conclude that reverse engineering and stochastic simulations are a useful approach to find optimal phage doses against pathogenic bacteria, an important step for the implementation of phage-therapy.

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