Discriminating typical and atypical cystic fibrosis‐related bacteria by multiplex PNA‐FISH

Lopes, Susana Patrícia; Carvalho, Daniel T. O.; Azevedo, Nuno F.

doi:10.1002/bit.26085

Cited by 15 publications

(14 citation statements)

References 98 publications

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“…However, using a decreasing inoculum to a level more clinically relevant will result in bleaching while trying to locate the few bacteria. Because PNA FISH is faster than the traditional culture methods used at the hospitals to day work is still done to optimize the use for CF patients .…”

Section: Application Methods and Practical Examples Of Fluorescent Smentioning

confidence: 99%

The use of fluorescent staining techniques for microscopic investigation of polymorphonuclear leukocytes and bacteria

Alhede

Stavnsbjerg

Bjarnsholt

2018

APMIS

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The use of fluorescent stains to visually investigate eukaryotic and/or prokaryotic cells is increasing quickly and manuscripts within all areas of research publish results using fluorescent staining techniques. However, in contrast to literature on traditional histological staining techniques, the literature on fluorescent stains and staining techniques does not offer as good an illustration of cellular morphology. The aim of this guideline is to illustrate different fluorescent stains and staining techniques for imaging immune cells, in particular the polymorphonuclear leukocytes (PMNs), in combination with infecting bacteria as seen in chronic bacterial infections. Thereby providing the first guideline for the morphology and identification of fluorescently stained PMNs, bacteria and biofilm.

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Section: Application Methods and Practical Examples Of Fluorescent Smentioning

confidence: 99%

The use of fluorescent staining techniques for microscopic investigation of polymorphonuclear leukocytes and bacteria

Alhede

Stavnsbjerg

Bjarnsholt

2018

APMIS

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“…aerugionosa . This PNA probe, designated as Paer565, was previously designed, optimized and validated on biofilms by Lopes et al [56]. Briefly, dual-species biofilms of P .…”

Section: Methodsmentioning

confidence: 99%

Polymicrobial Ventilator-Associated Pneumonia: Fighting In Vitro Candida albicans-Pseudomonas aeruginosa Biofilms with Antifungal-Antibacterial Combination Therapy

et al. 2017

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The polymicrobial nature of ventilator-associated pneumonia (VAP) is now evident, with mixed bacterial-fungal biofilms colonizing the VAP endotracheal tube (ETT) surface. The microbial interplay within this infection may contribute for enhanced pathogenesis and exert impact towards antimicrobial therapy. Consequently, the high mortality/morbidity rates associated to VAP and the worldwide increase in antibiotic resistance has promoted the search for novel therapeutic strategies to fight VAP polymicrobial infections. Under this scope, this work aimed to assess the activity of mono- vs combinational antimicrobial therapy using one antibiotic (Polymyxin B; PolyB) and one antifungal (Amphotericin B; AmB) agent against polymicrobial biofilms of Pseudomonas aeruginosa and Candida albicans. The action of isolated antimicrobials was firstly evaluated in single- and polymicrobial cultures, with AmB being more effective against C. albicans and PolyB against P. aeruginosa. Mixed planktonic cultures required equal or higher antimicrobial concentrations. In biofilms, only PolyB at relatively high concentrations could reduce P. aeruginosa in both monospecies and polymicrobial populations, with C. albicans displaying only punctual disturbances. PolyB and AmB exhibited a synergistic effect against P. aeruginosa and C. albicans mixed planktonic cultures, but only high doses (256 mg L-1) of PolyB were able to eradicate polymicrobial biofilms, with P. aeruginosa showing loss of cultivability (but not viability) at 2 h post-treatment, whilst C. albicans only started to be inhibited after 14 h. In conclusion, combination therapy involving an antibiotic and an antifungal agent holds an attractive therapeutic option to treat severe bacterial-fungal polymicrobial infections. Nevertheless, optimization of antimicrobial doses and further clinical pharmacokinetics/pharmacodynamics and toxicodynamics studies underpinning the optimal use of these drugs are urgently required to improve therapy effectiveness and avoid reinfection.

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“…However, it should be considered that antimicrobial therapies that kill bacteria without sufficiently compromising the cell membrane can result in overestimation of viable bacteria using PMA-qRT-PCR [94]. An alternative method for evaluating microbial community composition is PNA-FISH ( peptide nucleic acid-fluorescence in situ hybridisation) which can be combined with imaging, hence providing information on spatial organisation of communities [95]. LOPES et al [96] recently quantified bacteria in a multispecies biofilm consisting of P. aeruginosa, Inquilinus limosus and Dolosigranulum pigrum comparing plating on selective media, qRT-PCR and PNA-FISH.…”

Section: Microbial Community Membersmentioning

confidence: 99%

Influence of the lung microbiome on antibiotic susceptibility of cystic fibrosis pathogens

et al. 2019

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The lungs of patients with cystic fibrosis (CF) are colonised by a microbial community comprised of pathogenic species, such as Pseudomonas aeruginosa and Staphylococcus aureus, and microorganisms that are typically not associated with worse clinical outcomes (considered as commensals). Antibiotics directed at CF pathogens are often not effective and a discrepancy is observed between activity of these agents in vitro and in the patient. This review describes how interspecies interactions within the lung microbiome might influence the outcome of antibiotic treatment targeted at common CF pathogens. Protective mechanisms by members of the microbiome such as antibiotic degradation (indirect pathogenicity), alterations of the cell wall, production of matrix components decreasing antibiotic penetration, and changes in metabolism are discussed. Interspecies interactions that increase bacterial susceptibility are also addressed. Furthermore, we discuss how experimental conditions, such as culture media, oxygen levels, incorporation of host–pathogen interactions, and microbial community composition may influence the outcome of microbial interaction studies related to antibiotic activity. Hereby, the importance to create in vitro conditions reflective of the CF lung microenvironment is highlighted. Understanding the role of the CF lung microbiome in antibiotic efficacy may help find novel therapeutic and diagnostic approaches to better tackle chronic lung infections in this patient population.

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Discriminating typical and atypical cystic fibrosis‐related bacteria by multiplex PNA‐FISH

Cited by 15 publications

References 98 publications

The use of fluorescent staining techniques for microscopic investigation of polymorphonuclear leukocytes and bacteria

The use of fluorescent staining techniques for microscopic investigation of polymorphonuclear leukocytes and bacteria

Polymicrobial Ventilator-Associated Pneumonia: Fighting In Vitro Candida albicans-Pseudomonas aeruginosa Biofilms with Antifungal-Antibacterial Combination Therapy

Influence of the lung microbiome on antibiotic susceptibility of cystic fibrosis pathogens

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