scholarly journals Selective Eradication of Staphylococcus aureus by the Designer Genetically Programmed Yeast Biocontrol Agent

Antibiotics ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 527
Author(s):  
Sofiya O. Pipiya ◽  
Yuliana A. Mokrushina ◽  
Alexander G. Gabibov ◽  
Ivan V. Smirnov ◽  
Stanislav S. Terekhov
Keyword(s):  
Staphylococcus Aureus ◽  
Biocontrol Agent ◽  
Infectious Agent ◽  
Cost Effective ◽  
Biocontrol Agents ◽  
Alternative View ◽  
Ecological Niches ◽  
In Vivo Models ◽  
Conventional Antibiotics

Staphylococcus aureus is a common human pathogen that is particularly often associated with antibiotic resistance. The eradication of this ubiquitous infectious agent from its ecological niches and contaminated surfaces is especially complicated by excessive biofilm formation and persisting cells, which evade the antibacterial activity of conventional antibiotics. Here, we present an alternative view of the problem of specific S. aureus eradication. The constitutive heterologous production of highly specific bacteriolytic protease lysostaphin in yeast Pichia pastoris provides an efficient biocontrol agent, specifically killing S. aureus in coculture. A yeast-based anti-S. aureus probiotic was efficient in a high range of temperatures and target-to-effector ratios, indicating its robustness and versatility in eliminating S. aureus cells. The efficient eradication of S. aureus by live lysostaphin-producing P. pastoris was achieved at high scales, providing a simple, biocompatible and cost-effective strategy for S. aureus lysis in bioproduction and surface decontamination. Future biomedical applications based on designer yeast biocontrol agents require evaluation in in vivo models. However, we believe that this strategy is very promising since it provides highly safe, efficient and selective genetically programmed probiotics and targeted biocontrol agents.

scholarly journals Efficacy of NZ2114, a Novel Plectasin-Derived Cationic Antimicrobial Peptide Antibiotic, in Experimental Endocarditis Due to Methicillin-Resistant Staphylococcus aureus

2011 ◽  
Vol 55 (11) ◽  
pp. 5325-5330 ◽  
Author(s):  
Yan Q. Xiong ◽  
Wessam Abdel Hady ◽  
Antoine Deslandes ◽  
Astrid Rey ◽  
Laurent Fraisse ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Maximum Effect ◽  
Peptide Antibiotic ◽  
Time Curve ◽  
Cationic Antimicrobial Peptide ◽  
Unbound Fraction ◽  
Methicillin Resistant ◽  
Content Type ◽  
Conventional Antibiotics

ABSTRACTCationic antimicrobial peptides (CAPs) play important roles in host immune defenses. Plectasin is a defensin-like CAP isolated from the saprophytic fungusPseudoplectania nigrella. NZ2114 is a novel variant of plectasin with potent activity against Gram-positive bacteria. In this study, we investigated (i) thein vivopharmacokinetic and pharmacodynamic (PK/PD) characteristics of NZ2114 and (ii) thein vivoefficacy of NZ2114 in comparison with those of two conventional antibiotics, vancomycin or daptomycin, in an experimental rabbit infective endocarditis (IE) model due to a methicillin-resistantStaphylococcus aureus(MRSA) strain (ATCC 33591). All NZ2114 regimens (5, 10, and 20 mg/kg of body weight, intravenously [i.v.], twice daily for 3 days) significantly decreased MRSA densities in cardiac vegetations, kidneys, and spleen versus those in untreated controls, except in one scenario (5 mg/kg, splenic MRSA counts). The efficacy of NZ2114 was clearly dose dependent in all target tissues. At 20 mg/kg, NZ2114 showed a significantly greater efficacy than vancomycin (P< 0.001) and an efficacy similar to that of daptomycin. Of importance, only NZ2114 (in 10- and 20-mg/kg regimens) prevented posttherapy relapse in cardiac vegetations, kidneys, and spleen, while bacterial counts in these target tissues continued to increase in vancomycin- and daptomycin-treated animals. Thesein vivoefficacies were equivalent and significantly correlated with three PK indices investigated:fCmax/MIC (the maximum concentration of the free, unbound fraction of a drug in serum divided by the MIC),fAUC/MIC (where AUC is the area under the concentration-time curve), andf%T>MIC(%T>MICis the cumulative percentage of a 24-h period that the drug concentration exceeds the MIC under steady-state pharmacokinetic conditions), as analyzed by a sigmoid maximum-effect (Emax) model (R2> 0.69). The superior efficacy of NZ2114 in this MRSA IE model suggests the potential for further development of this compound for treating serious MRSA infections.

scholarly journals Clearance of Staphylococcus aureus from In Vivo Models of Chronic Infection by Immunization Requires Both Planktonic and Biofilm Antigens

2019 ◽  
Vol 88 (1) ◽  
Author(s):  
Janette M. Harro ◽  
Yvonne Achermann ◽  
Jeffrey A. Freiberg ◽  
Devon L. Allison ◽  
Kristen J. Brao ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Protective Efficacy ◽  
Passive Immunization ◽  
Quadrivalent Vaccine ◽  
Bacterial Populations ◽  
In Vivo Models ◽  
Content Type ◽  
Pentavalent Vaccine ◽  
And Control

ABSTRACT Staphylococcus aureus is a causative agent of chronic biofilm-associated infections that are recalcitrant to resolution by the immune system or antibiotics. To combat these infections, an antistaphylococcal, biofilm-specific quadrivalent vaccine against an osteomyelitis model in rabbits has previously been developed and shown to be effective at eliminating biofilm-embedded bacterial populations. However, the addition of antibiotics was required to eradicate remaining planktonic populations. In this study, a planktonic upregulated antigen was combined with the quadrivalent vaccine to remove the need for antibiotic therapy. Immunization with this pentavalent vaccine followed by intraperitoneal challenge of BALB/c mice with S. aureus resulted in 16.7% and 91.7% mortality in pentavalent vaccine and control groups, respectively (P < 0.001). Complete bacterial elimination was found in 66.7% of the pentavalent cohort, while only 8.3% of the control animals cleared the infection (P < 0.05). Further protective efficacy was observed in immunized rabbits following intramedullary challenge with S. aureus, where 62.5% of the pentavalent cohort completely cleared the infection, versus none of the control animals (P < 0.05). Passive immunization of BALB/c mice with serum IgG against the vaccine antigens prior to intraperitoneal challenge with S. aureus prevented mortality in 100% of mice and eliminated bacteria in 33.3% of the challenged mice. These results demonstrate that targeting both the planktonic and biofilm stages with the pentavalent vaccine or the IgG elicited by immunization can effectively protect against S. aureus infection.

scholarly journals Impact of Bicarbonate-β-Lactam Exposures on Methicillin-Resistant Staphylococcus aureus (MRSA) Gene Expression in Bicarbonate-β-Lactam-Responsive vs. Non-Responsive Strains

Genes ◽  
2021 ◽  
Vol 12 (11) ◽  
pp. 1650
Author(s):  
Selvi C. Ersoy ◽  
Blake M. Hanson ◽  
Richard A. Proctor ◽  
Cesar A. Arias ◽  
Truc T. Tran ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Ex Vivo ◽  
In Vivo Models ◽  
Methicillin Resistant ◽  
Genetic Perturbations ◽  
Functional Relevance

Methicillin-resistant Staphylococcus aureus (MRSA) infections represent a difficult clinical treatment issue. Recently, a novel phenotype was discovered amongst selected MRSA which exhibited enhanced β-lactam susceptibility in vitro in the presence of NaHCO3 (termed ‘NaHCO3-responsiveness’). This increased β-lactam susceptibility phenotype has been verified in both ex vivo and in vivo models. Mechanistic studies to-date have implicated NaHCO3-mediated repression of genes involved in the production, as well as maturation, of the alternative penicillin-binding protein (PBP) 2a, a necessary component of MRSA β-lactam resistance. Herein, we utilized RNA-sequencing (RNA-seq) to identify genes that were differentially expressed in NaHCO3-responsive (MRSA 11/11) vs. non-responsive (COL) strains, in the presence vs. absence of NaHCO3-β-lactam co-exposures. These investigations revealed that NaHCO3 selectively repressed the expression of a cadre of genes in strain 11/11 known to be a part of the sigB-sarA-agr regulon, as well as a number of genes involved in the anchoring of cell wall proteins in MRSA. Moreover, several genes related to autolysis, cell division, and cell wall biosynthesis/remodeling, were also selectively impacted by NaHCO3-OXA exposure in the NaHCO3-responsive strain MRSA 11/11. These outcomes provide an important framework for further studies to mechanistically verify the functional relevance of these genetic perturbations to the NaHCO3-responsiveness phenotype in MRSA.

scholarly journals Animal models in bicompatibility assessments of implants in soft and hard tissues

2021 ◽  
pp. 5-5
Author(s):  
Bogomir Prokic ◽  
Tijana Luzajic-Bozinovski ◽  
Vladimir Gajdov ◽  
Ivan Milosevic ◽  
Vera Todorovic ◽  
...  
Keyword(s):  
Cost Effective ◽  
Screening Tests ◽  
Laboratory Animals ◽  
In Vivo Models ◽  
Technological Advances ◽  
Hard Tissues ◽  
In Vivo Testing ◽  
Tissue Recovery ◽  
Eukaryotic Organisms

The ethical dilemmas of using animals as in vivo models in preclinical and clinical examinations have been increasingly present in recent decades. Small laboratory animals (rats, rabbits) will continue to be used because they are cost-effective and permit the formation of statistically testable cohort groups; a task that, for financial, maintenance and care reasons, is almost prohibitive for larger animals. Technological advances in the production of new biomaterials for clinical use are enormous, but screening tests and methods used to assess biocompatibility lag behind these advances. The assessment of biological responses is slow and based on millennial recovery mechanisms in eukaryotic organisms. Therefore, the goal of researchers in this field is to re-evaluate old methods of biocompatibility assessment and introduce new methods of evaluation, especially for in vivo testing. In that sense, a revision of the ISO standards was planned and conducted in 2017, which insisted on cytotoxicity testing in cell lines and produced concrete proposals on how biocompatibility should be quantified. In vivo biocompatibility evaluation of biomaterials used for soft tissue recovery commonly utilises rats. Rabbits are recommended for implants used for hard tissues, because of the rabbit?s size, the possibility of implanting the biomaterials on a larger bone surface, and because of the peculiarities of rabbit bone tissue that favours rapid recovery after bone defects and enables easy reading of the results.

scholarly journals A Strain of Bacillus amyloliquefaciens Can Prevent Vibrio vulnificus Colonization in Crassostrea gigas Oysters

2020 ◽  
Vol 7 ◽  
Author(s):  
Andrea Martha Freire-Peñaherrera ◽  
Ana Tirapé ◽  
Jerry Landívar-Zambrano ◽  
Juan Manuel Cevallos-Cevallos
Keyword(s):  
Crassostrea Gigas ◽  
Bacillus Amyloliquefaciens ◽  
Biocontrol Agent ◽  
Vibrio Vulnificus ◽  
Biocontrol Agents ◽  
Biocontrol Efficacy ◽  
Solid Media ◽  
Clinical Cases

The pathogen Vibrio vulnificus has been associated with the majority of clinical cases of septicemia and deaths attributed to shellfish consumption. However, reports on biocontrol agents against this pathogen are scarce. In this study, the strain A5 of Bacillus amyloliquefaciens (A5) was evaluated against V. vulnificus. The sensitivity of V. vulnificus to A5 was first assessed in vitro using selected solid media as well as autoclaved oysters (Crassostrea gigas). Then, the ability of A5 to colonize live oysters was evaluated, and the biocontrol efficacy was investigated in vivo using oysters inoculated with V. vulnificus before or after inoculation with A5. The survival of the pathogen in oysters was evaluated after 2, 3, 4, and 6 days of exposure to A5 in all the experiments. In vitro, A5 showed inhibition halos of 18 mm against V. vulnificus. In autoclaved oysters, A5 caused a significant reduction in the levels of V. vulnificus on day 2 at 5.14 log CFU/g, but the pathogen’s counts were restored after day 3. In vivo, A5 was able to survive in live oysters and prevented the colonization of V. vulnificus only when the biocontrol agent was inoculated before the pathogen. Results show the potential of A5 to prevent V. vulnificus uptake by oysters when administered prior to the pathogen.

scholarly journals Activities of Clindamycin, Daptomycin, Doxycycline, Linezolid, Trimethoprim-Sulfamethoxazole, and Vancomycin against Community-Associated Methicillin-Resistant Staphylococcus aureus with Inducible Clindamycin Resistance in Murine Thigh Infection and In Vitro Pharmacodynamic Models

2008 ◽  
Vol 52 (6) ◽  
pp. 2156-2162 ◽  
Author(s):  
Kerry L. LaPlante ◽  
Steven N. Leonard ◽  
David R. Andes ◽  
William A. Craig ◽  
Michael J. Rybak
Keyword(s):  
Staphylococcus Aureus ◽  
Inoculum Size ◽  
In Vivo Model ◽  
Infection Model ◽  
Time Curve ◽  
In Vivo Models ◽  
Methicillin Resistant ◽  
Inducible Clindamycin Resistance

ABSTRACT Controversy exists about the most effective treatment options for community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) and about the ability of these strains to develop inducible resistance to clindamycin during therapy. Using both in vitro pharmacodynamic and murine thigh infection models, we evaluated and compared several antimicrobial compounds against CA-MRSA. Strains with inducible macrolide lincosamide-streptogramin type B (iMLSB) resistance and strains in which resistance was noninducible were evaluated. Two levels of inocula (105 and 107) were evaluated for clindamycin activity in the in vivo model. In both models, the antimicrobial evaluation was performed in triplicate, and bacterial quantification occurred over 72 h, with drug doses that were designed to simulate the free drug area-under-the-concentration-time curve values (fAUCs) obtained from human samples. When the activity of clindamycin against the iMLSB strains was evaluated, constitutive resistance was noted at 24 h (MIC of >256), and failure was noted at an inoculum of ≥106 in the in vivo models. However, at a low inoculum (105) in the murine thigh-infection model, clindamycin demonstrated modest activity, reducing the CFU/thigh count for clindamycin resistance-inducible strains at 72 h (0.45 to 1.3 logs). Overall, administration of daptomycin followed by vancomycin demonstrated the most significant kill against all strains in both models. Against the clindamycin noninducible strain, clindamycin and doxycycline demonstrated significant kill. Doxycycline, linezolid, and trimethoprim-sulfamethoxazide (not run in the murine model) demonstrated bacteriostatic activity against clindamycin resistance-inducible isolates. This study demonstrates that clindamycin's activity against the iMLSB strains tested is partially impacted by inoculum size. At present, there are several alternatives that appear promising for treating clindamycin resistance-inducible strains of CA-MRSA.

scholarly journals Lipidation of Temporin-1CEb Derivatives as a Tool for Activity Improvement, Pros and Cons of the Approach

2021 ◽  
Vol 22 (13) ◽  
pp. 6679
Author(s):  
Paulina Kosikowska-Adamus ◽  
Emilia Sikorska ◽  
Dariusz Wyrzykowski ◽  
Aleksandra Walewska ◽  
Anna Golda ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Peptides ◽  
Antimicrobial Properties ◽  
Carbon Chain ◽  
Microbial Pathogens ◽  
Multi Drug Resistance ◽  
Aggregation State ◽  
Conventional Antibiotics ◽  
Biofilm Development

The alarming raise of multi-drug resistance among human microbial pathogens makes the development of novel therapeutics a priority task. In contrast to conventional antibiotics, antimicrobial peptides (AMPs), besides evoking a broad spectrum of activity against microorganisms, could offer additional benefits, such as the ability to neutralize toxins, modulate inflammatory response, eradicate bacterial and fungal biofilms or prevent their development. The latter properties are of special interest, as most antibiotics available on the market have limited ability to diffuse through rigid structures of biofilms. Lipidation of AMPs is considered as an effective approach for enhancement of their antimicrobial potential and in vivo stability; however, it could also have undesired impact on selectivity, solubility or the aggregation state of the modified peptides. In the present work, we describe the results of structural modifications of compounds designed based on cationic antimicrobial peptides DK5 and CAR-PEG-DK5, derivatized at their N-terminal part with fatty acids with different lengths of carbon chain. The proposed modifications substantially improved antimicrobial properties of the final compounds and their effectiveness in inhibition of biofilm development as well as eradication of pre-formed 24 h old biofilms of Candida albicans and Staphylococcus aureus. The most active compounds (C5-DK5, C12-DK5 and C12-CAR-PEG-DK5) were also potent against multi-drug resistant Staphylococcus aureus USA300 strain and clinical isolates of Pseudomonas aeruginosa. Both experimental and in silico methods revealed strong correlation between the length of fatty acid attached to the peptides and their final membranolytic properties, tendency to self-assemble and cytotoxicity.

scholarly journals Comprehensive Evaluation of the Toxicity and Biosafety of Plasma Polymerized Nanoparticles

Nanomaterials ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1176
Author(s):  
Praveesuda L. Michael ◽  
Yuen Ting Lam ◽  
Juichien Hung ◽  
Richard P. Tan ◽  
Miguel Santos ◽  
...  
Keyword(s):  
Comprehensive Evaluation ◽  
Cost Effective ◽  
Cell Types ◽  
Repeated Injection ◽  
In Vivo Models ◽  
Intravenous Injections ◽  
Cost Effective Alternative ◽  
High Doses

The rapid growth of nanoparticle-based therapeutics has underpinned significant developments in nanomedicine, which aim to overcome the limitations imposed by conventional therapies. Establishing the safety of new nanoparticle formulations is the first important step on the pathway to clinical translation. We have recently shown that plasma-polymerized nanoparticles (PPNs) are highly efficient nanocarriers and a viable, cost-effective alternative to conventional chemically synthesized nanoparticles. Here, we present the first comprehensive toxicity and biosafety study of PPNs using both established in vitro cell models and in vivo models. Overall, we show that PPNs were extremely well tolerated by all the cell types tested, significantly outperforming commercially available lipid-based nanoparticles (lipofectamine) used at the manufacturer’s recommended dosage. Supporting the in vitro data, the systemic toxicity of PPNs was negligible in BALB/c mice following acute and repeated tail-vein intravenous injections. PPNs were remarkably well tolerated in mice without any evidence of behavioral changes, weight loss, significant changes to the hematological profile, or signs of histological damage in tissues. PPNs were tolerated at extremely high doses without animal mortality observed at 6000 mg/kg and 48,000 mg/kg for acute and repeated-injection regimens, respectively. Our findings demonstrate the safety of PPNs in biological systems, adding to their future potential in biomedical applications.

scholarly journals AMP–Chitosan Coating with Bactericidal Activity in the Presence of Human Plasma Proteins

Molecules ◽  
2020 ◽  
Vol 25 (13) ◽  
pp. 3046 ◽  
Author(s):  
Cláudia Monteiro ◽  
Hélia Fernandes ◽  
Diana Oliveira ◽  
Nuno Vale ◽  
Mariana Barbosa ◽  
...  
Keyword(s):  
Plasma Proteins ◽  
Cost Effective ◽  
Wound Dressings ◽  
Advanced Materials ◽  
Efficient Treatment ◽  
Chitosan Coating ◽  
Bactericidal Properties ◽  
Improved Performance ◽  
Conventional Antibiotics

Antibiotic resistance is increasing and new strategies are needed to fight infection. Advanced materials are promising tools that can be combined with innovative alternatives to conventional antibiotics to allow more targeted and efficient treatment. In this work, we explored the activity against Staphylococcus epidermidis (S. epidermidis) of the α-helical antimicrobial peptide (AMP) MSI-78(4-20) (KFLKKAKKFGKAFVKIL) when covalently bound to a chitosan coating. The AMP MSI-78(4-20) (17 mer) is an improved version of its parent MSI-78 (22 mer; commercially known as Pexiganan), a cost-effective short AMP, which was demonstrated to be as effective as MSI-78 and less toxic to eukaryotic cells. An MSI-78(4-20)–chitosan coating could be applied in several infection scenarios, ranging from bone implants to wound dressings, as chitosan possesses osteoconductive and hemostatic properties. Cysteine-modified MSI-78(4-20) was covalently immobilized onto the chitosan coating through a succinimidyl-[(N-maleimidopropionamido)-octaethyleneglycol] ester (SM(PEG)8), a heterobifuncional crosslinker, with N-hydroxysuccinimide (NHS) ester and maleimide groups, by its N- and C- termini. The MSI-78(4-20)–chitosan coating demonstrated bactericidal properties independently of the tethering site and an improved performance in the presence of plasma proteins, which mimics conditions that will be encountered in vivo. This AMP–chitosan coating has therefore great potential for applications in medical devices such as implants or even wound dressings.

Sign in / Sign up

Export Citation Format

Share Document