Discrimination of bacteriophage infected cells using locked nucleic acid fluorescent <i>in situ</i> hybridization (LNA-FISH)

Boas, Diana Patrícia Andrade Vilas; Almeida, Carina; Sillankorva, Sanna; Nicolau, Ana; Azeredo, Joana; Azevedo, Nuno F.

doi:10.1080/08927014.2015.1131821

Cited by 29 publications

(17 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Bacteriophages are species specific, and so can be used to target pathogenic bacteria, without disturbing non-harmful commensal bacteria (Hanlon, 2007 ). Bacteriophages are also able to penetrate bacterial biofilms (Vilas Boas et al, 2016 ).…”

Section: Introductionmentioning

confidence: 99%

Activity of Bacteriophages in Removing Biofilms of Pseudomonas aeruginosa Isolates from Chronic Rhinosinusitis Patients

Fong

Drilling

Morales³

et al. 2017

Front. Cell. Infect. Microbiol.

129

119

View full text Add to dashboard Cite

Introduction: Pseudomonas aeruginosa infections are prevalent amongst chronic rhinosinusitis (CRS) sufferers. Many P. aeruginosa strains form biofilms, leading to treatment failure. Lytic bacteriophages (phages) are viruses that infect, replicate within, and lyse bacteria, causing bacterial death.Aim: To assess the activity of a phage cocktail in eradicating biofilms of ex vivo P.aeruginosa isolates from CRS patients.Methods: P. aeruginosa isolates from CRS patients with and without cystic fibrosis (CF) across three continents were multi-locus sequence typed and tested for antibiotic resistance. Biofilms grown in vitro were treated with a cocktail of four phages (CT-PA). Biofilm biomass was measured after 24 and 48 h, using a crystal violet assay. Phage titrations were performed to confirm replication of the phages. A linear mixed effects model was applied to assess the effects of treatment, time, CF status, and multidrug resistance on the biomass of the biofilm.Results: The isolates included 44 strain types. CT-PA treatment significantly reduced biofilm biomass at both 24 and 48 h post-treatment (p < 0.0001), regardless of CF status or antibiotic resistance. Biomass was decreased by a median of 76% at 48 h. Decrease in biofilm was accompanied by a rise in phage titres for all except one strain.Conclusion: A single dose of phages is able to significantly reduce biofilms formed in vitro by a range of P.aeruginosa isolates from CRS patients. This represents an exciting potential and novel targeted treatment for P. aeruginosa biofilm infections and multidrug resistant bacteria.

show abstract

Section: Introductionmentioning

confidence: 99%

Activity of Bacteriophages in Removing Biofilms of Pseudomonas aeruginosa Isolates from Chronic Rhinosinusitis Patients

Fong

Drilling

Morales³

et al. 2017

Front. Cell. Infect. Microbiol.

129

119

View full text Add to dashboard Cite

show abstract

“…Like all viruses, bacteriophages are highly species‐specific with regard to their hosts, and thus can be used to target pathogenic bacteria without disturbing commensal bacteria. The additional capacity of bacteriophages to penetrate bacterial biofilms has made them an attractive nonantibiotic topical therapy for treating bacterial biofilms in CRS.…”

Section: Resultsmentioning

confidence: 99%

“…As infectivity is species and sometimes strain‐limited, the use of phage cocktails rather than individual phages has been shown to improve activity by expanding the host range and by preventing the development of bacteriophage‐insensitive mutant bacteria . Moreover, advances in biotechnology and synthetic biology fields have enabled the development of a new generation of phages that are more effective at controlling biofilms …”

Section: Resultsmentioning

confidence: 99%

“…Although further evaluation and optimization of phage cocktails in the treatment of CRS are required, the safety profile of topical phage therapy and its activity against antibiotic‐resistant isolates make it an attractive potential therapeutic option. Preclinical and clinical studies will be of great benefit in determining whether phage therapy can fulfill its potential and meet its expectations . However, the long‐term success of phage therapy will be predicated on the development of new strategies to overcome the current limitations of bacteriophages as therapeutic agents, the creation of an adequate regulatory framework, the implementation of safety protocols, and acceptance by the general public …”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Future topical medications in chronic rhinosinusitis

Miyake

Bleier

2019

Int Forum Allergy Rhinol

View full text Add to dashboard Cite

Background:Research has progressed rapidly in recent decades to be er understand the etiopathogenesis and management paradigms of chronic rhinosinusitis (CRS). Although oral antibiotics o en mitigate symptoms in acute CRS exacerbations, eradication of polymicrobial biofilms and multidrug-resistant bacteria remains a challenge. The goal of this review is to summarize and discuss the potential and pitfalls of topical medications in the treatment of CRS. Methods:A related literature review was performed using PubMed and Scopus, with only the English database included. Results:The main therapies were selected and separated in sections. Details regarding future topical treatments of CRS were summarized and discussed. Conclusion:The ease of access of the sinonasal mucosa positions CRS as a disease with high potential for local topical treatment. The ultimate adoption of topical agents will require continued expansion of our understanding of novel local targets in CRS as well as improved methods to deliver and retain the drug of interest at the site of activity. C 2019 ARS-AAOA, LLC.

show abstract

“…Lytic spectrum of phages isolated against clinical isolates of A. baumannii. wrapped in aluminium foil and incubated at a hybridization temperature of 59°C for 90 min (Almeida et al, 2009;Vilas Boas et al, 2016). After, the coverslip was removed and slides washed with pre-heated specific wash solutions (WS) during 30 min at 59°C to remove unbound probes.…”

Section: Tablementioning

confidence: 99%

Antimicrobial assessment of phage therapy using a porcine model of biofilm infection

Milho

Andrade

Boas

et al. 2019

International Journal of Pharmaceutics

View full text Add to dashboard Cite

Antibiotic resistant bacterial communities persist in many types of wounds, chronic wounds in particular, in the form of biofilms. Biofilm formation is a major cause of severe infections and the main reason for a negative treatment outcome and slow healing progression. Chronic wounds are a silent epidemic essentially affecting people with co-morbid conditions such as diabetes and obesity and elderly persons particularly those with movement limitations. The development of complementary and alternative effective strategies to antibiotics for the treatment of chronic wounds is highly desired. Phage therapy constitutes a very promising approach in the control of topical microbial populations. In this work newly isolated phages were tested for their efficacy to control bacterial species that predominate in chronic wounds. Phage effectiveness was studied on 24-h old biofilms formed in polystyrene microplates and in porcine skin explants using two treatment approaches: individual phage and a cocktail of phages against four main pathogens commonly isolated from chronic wounds. The two models produced variations in the surface colonization ability, assessed by viable bacterial counts and microscopy visualization after using peptide nucleic acid (PNA) or locked nucleic acid probes (LNA) and 2′-Omethyl (2′-OMe) in fluorescence in situ hybridization (FISH), and in the phage-host interactions. Phages alone and combined caused greater reductions in the number of viable cells when biofilms had been formed on porcine skins and with greater variations detected at 4 h and 24 h of sampling. These results suggest that porcine skin models should be preferentially used to assess the use of phages and phage cocktails intended for topical use in order to understand the fate, throughout treatment time, of the population when dealing with biofilm-related infections.

show abstract

Discrimination of bacteriophage infected cells using locked nucleic acid fluorescent in situ hybridization (LNA-FISH)

Cited by 29 publications

References 48 publications

Activity of Bacteriophages in Removing Biofilms of Pseudomonas aeruginosa Isolates from Chronic Rhinosinusitis Patients

Activity of Bacteriophages in Removing Biofilms of Pseudomonas aeruginosa Isolates from Chronic Rhinosinusitis Patients

Future topical medications in chronic rhinosinusitis

Antimicrobial assessment of phage therapy using a porcine model of biofilm infection

Contact Info

Product

Resources

About