scholarly journals Human organoid biofilm model for assessing antibiofilm activity of novel agents

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Bing (Catherine) Wu ◽  
Evan F. Haney ◽  
Noushin Akhoundsadegh ◽  
Daniel Pletzer ◽  
Michael J. Trimble ◽  
...  

AbstractBacterial biofilms cause 65% of all human infections and are highly resistant to antibiotic therapy but lack specific treatments. To provide a human organoid model for studying host-microbe interplay and enabling screening for novel antibiofilm agents, a human epidermis organoid model with robust methicillin-resistant Staphylococcus aureus (MRSA) USA300 and Pseudomonas aeruginosa PAO1 biofilm was developed. Treatment of 1-day and 3-day MRSA and PAO1 biofilms with antibiofilm peptide DJK-5 significantly and substantially reduced the bacterial burden. This model enabled the screening of synthetic host defense peptides, revealing their superior antibiofilm activity against MRSA compared to the antibiotic mupirocin. The model was extended to evaluate thermally wounded skin infected with MRSA biofilms resulting in increased bacterial load, cytotoxicity, and pro-inflammatory cytokine levels that were all reduced upon treatment with DJK-5. Combination treatment of DJK-5 with an anti-inflammatory peptide, 1002, further reduced cytotoxicity and skin inflammation.

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Agnieszka Latka ◽  
Zuzanna Drulis-Kawa

AbstractOne of the potential antibiofilm strategies is to use lytic phages and phage-derived polysaccharide depolymerases. The idea is to uncover bacteria embedded in the biofilm matrix making them accessible and vulnerable to antibacterials and the immune system. Here we present the antibiofilm efficiency of lytic phage KP34 equipped with virion-associated capsule degrading enzyme (depolymerase) and its recombinant depolymerase KP34p57, depolymerase-non-bearing phage KP15, and ciprofloxacin, separately and in combination, using a multidrug-resistant K. pneumoniae biofilm model. The most effective antibiofilm agents were (1) phage KP34 alone or in combination with ciprofloxacin/phage KP15, and (2) depolymerase KP34p57 with phage KP15 and ciprofloxacin. Secondly, applying the commonly used biofilm microtiter assays: (1) colony count, (2) LIVE/DEAD BacLight Bacterial Viability Kit, and (3) crystal violet (CV) biofilm staining, we unravelled the main advantages and limitations of the above methods in antibiofilm testing. The diverse mode of action of selected antimicrobials strongly influenced obtained results, including a false positive enlargement of biofilm mass (CV staining) while applying polysaccharide degrading agents. We suggest that to get a proper picture of antimicrobials’ effectiveness, multiple examination methods should be used and the results must be read considering the principle of each technique and the antibacterial mechanism.


Antibiotics ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 77 ◽  
Author(s):  
Virginio Cepas ◽  
Yuly López ◽  
Yaiza Gabasa ◽  
Clara B. Martins ◽  
Joana D. Ferreira ◽  
...  

Bacterial biofilms are complex biological systems that are difficult to eradicate at a medical, industrial, or environmental level. Biofilms confer bacteria protection against external factors and antimicrobial treatments. Taking into account that about 80% of human infections are caused by bacterial biofilms, the eradication of these structures is a great priority. Biofilms are resistant to old-generation antibiotics, which has led to the search for new antimicrobials from different sources, including deep oceans/seas. In this study, 675 extracts obtained from 225 cyanobacteria and microalgae species (11 phyla and 6 samples belonging to unknown group) were obtained from different culture collections: The Blue Biotechnology and Ecotoxicology Culture Collection (LEGE-CC), the Coimbra Collection of Algae (ACOI) from Portugal, and the Roscoff Culture Collection (RCC) from France. The largest number of samples was made up of the microalgae phylum Chlorophyta (270) followed by Cyanobacteria (261). To obtain a large range of new bioactive compounds, a method involving three consecutive extractions (hexane, ethyl acetate, and methanol) was used. The antibiofilm activity of extracts was determined against seven different bacterial species and two Candida strains in terms of minimal biofilm inhibitory concentration (MBIC). The highest biofilm inhibition rates (%) were achieved against Candida albicans and Enterobacter cloacae. Charophyta, Chlorophyta, and Cyanobacteria were the most effective against all microorganisms. In particular, extracts of Cercozoa phylum presented the lowest MBIC50 and MBIC90 values for all the strains except C. albicans.


2020 ◽  
Vol 17 (6) ◽  
pp. 466-471
Author(s):  
Usama W. Hawas ◽  
Fekri Shaher ◽  
Mohamed Ghandourah ◽  
Lamia T. Abou El-Kassem ◽  
Sathianeson Satheesh ◽  
...  

This study aimed at evaluating the antibiofilm activity of the Red Sea metabolites from green alga Avrainvillea amadelpha, sea cucumber Holothuria atra and costal plant Sarcocornia fruticosa against three biofilm bacterial strains isolated from Jeddah coast. Free fatty acids (FFAs) and other lipoidal matters were extracted from these organisms and analyzed by GC-MS. The composition of lipoidal fractions showed that A. amadelpha is rich by 74% saturated FAs, while sea cucumber H. atra revealed high content (60%) of unsaturated FAs. Palmitic acid is the major FA component in all species ranging from 14.5 to 26.7%. Phytol, sterols and hydrocarbons (C8-C29) were represented in the alga A. amadelpha as high contents with values 25.8, 21.9 and 18.5%, respectively. The extracts and lipoidal contents showed biofilm inhibitory activity against the isolated bacterial strains, where the unsaponified lipoidal fraction of S. fruticosa exhibited highest inhibitory activity against Planomicrobium sp. at concentration of 200 µg/mL.


2021 ◽  
Vol 9 (2) ◽  
pp. 450
Author(s):  
Maigualida Cuenca ◽  
María Carmen Sánchez ◽  
Pedro Diz ◽  
Lucía Martínez-Lamas ◽  
Maximiliano Álvarez ◽  
...  

The aim of this study was to evaluate the potential anti-biofilm and antibacterial activities of Streptococcus downii sp. nov. To test anti-biofilm properties, Streptococcus mutans, Actinomyces naeslundii, Veillonella parvula, Fusobacterium nucleatum, Porphyromonas gingivalis, and Aggregatibacter actinomycetemcomitans were grown in a biofilm model in the presence or not of S. downii sp. nov. for up to 120 h. For the potential antibacterial activity, 24 h-biofilms were exposed to S. downii sp. nov for 24 and 48 h. Biofilms structures and bacterial viability were studied by microscopy, and the effect in bacterial load by quantitative polymerase chain reaction. A generalized linear model was constructed, and results were considered as statistically significant at p < 0.05. The presence of S. downii sp. nov. during biofilm development did not affect the structure of the community, but an anti-biofilm effect against S. mutans was observed (p < 0.001, after 96 and 120 h). For antibacterial activity, after 24 h of exposure to S. downii sp. nov., counts of S. mutans (p = 0.019) and A. actinomycetemcomitans (p = 0.020) were significantly reduced in well-structured biofilms. Although moderate, anti-biofilm and antibacterial activities of S. downii sp. nov. against oral bacteria, including some periodontal pathogens, were demonstrated in an in vitro biofilm model.


2021 ◽  
Vol 0 (0) ◽  
pp. 0-0
Author(s):  
C.A. Lux ◽  
K. Biswas ◽  
M.W. Taylor ◽  
R.G. Douglas

Background: Despite best medical and surgical practice, some cases of chronic rhinosinusitis (CRS) can remain recalcitrant. Bacterial biofilms have been associated with the recalcitrance of sinonasal inflammation. Biofilms are highly resistant to commonly prescribed antibiotics. Accordingly, more effective antimicrobial treatment options are needed to treat refractory CRS. The aim of this study was to determine the in vitro efficacy of neutral electrolysed water (NEW) and povidone-iodine (PVI) against CRS-associated Staphylococcus aureus biofilms. Methods: Mature S. aureus biofilms were grown in a Centre for Disease Control (CDC) biofilm reactor. The antimicrobial activity of NEW, PVI and doxycycline was determined for both planktonic and biofilm cultures of a clinical S. aureus isolate using minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and minimum biofilm eradication concentration (MBEC) assays. Results: MICs and MBCs were determined for all antimicrobials. MBC values were similar to MICs for both antiseptics, but doxycycline MBCs were significantly higher than the associated MICs. Biofilms were highly resistant to NEW and doxycycline. The MBEC for doxycycline was between 500 and 1000 µg/mL. NEW was ineffective against biofilms and no MBEC could be determined. In contrast, a concentration of 10% of the commercial PVI solution (10 mg/mL PVI) led to effective eradication of mature biofilms. Conclusion: In this study, only PVI showed promising antibiofilm activity at physiological concentrations. The in vivo efficacy of PVI warrants further investigation of its potential as a treatment for recalcitrant CRS.


2018 ◽  
Vol 2 (2) ◽  
Author(s):  
Giuseppe Nebbioso ◽  
Ciro Falasconi ◽  
Viviana Nebbioso ◽  
Francesco Petrella

L’insorgenza di infezione su una lesione cutanea cronica determina un arresto del processo di riparazione tessutale e impone l’instaurazione di una terapia antibiotica sistemica che, in una fase iniziale, sarà empirica e, dunque, non scevra di insuccessi. L’utilizzo di antimicrobici con ridotto potere citotossoco/istiolesivo può essere considerata, in molti casi, una valida alternativa per il controllo della carica batterica e dell’infezione locale. L’associazione di antimicrobici a base di poliesanide biguanide, betaina e cadexomero iodico, in molti casi, permette di controllare/ridurre la carica batterica e l’infezione locale fungendo da starter per la ripresa del processo di riparazione tessutale. The onset of infection on a skin ulcer (chronic wound) leads to a halt in the tissue repair process and requires a systemic antibiotic therapy which, at an early stage, will be empirical and, therefore, not free from setbacks. The use of antimicrobials with reduced cytotoxic/histiolesive power can be considered, in many cases, a valid alternative for bacterial burden and local infection control. The combination of antimicrobials based on polystyrene biguanide, betaine and iodine cadexomer, in many cases, allows to control/reduce the bacterial burden and local infection by acting as a starter for the resumption of the tissue repair process.


2021 ◽  
Author(s):  
Emma Haley ◽  
Mederbek Matmusaev ◽  
Imtiyaz N. Hossain ◽  
Sean Davin ◽  
Tammy M. Martin ◽  
...  

AbstractBackgroundOverexpression of IL-23 in adult mice by means of hydrodynamic tail vein injection of IL-23 minicircles has been reported to result in spondyloarthritis-like disease. The impact of genetic background and sex on the disease phenotype in this model has not been investigated.MethodsWe compared male B10.RIII mice with male C57BL/6 mice, and male with female B10.RIII mice after hydrodynamic injection of IL-23 enhanced episomal vector (EEV) at 8-12 weeks of age. We monitored clinical arthritis scores, paw swelling, and body weight. Animals were euthanized after two weeks and tissues were harvested for histology, flow cytometry and gene expression analysis. Serum cytokine levels were determined by ELISA.FindingsMale B10.RIII mice developed arthritis in the forepaws and feet within 6 days after IL-23 EEV injection; they also exhibited psoriasis-like skin disease, colitis, weight loss, and osteopenia. In contrast to previous reports, we did not observe spondylitis or uveitis. Male C57BL/6 mice injected with IL-23 EEV had serum IL-23 levels comparable with B10.RIII mice and developed skin inflammation, colitis, weight loss, and osteopenia but failed to develop arthritis. Female B10.RIII mice had more severe arthritis than male B10.RIII mice but did not lose weight.ConclusionsSystemic IL-23 overexpression results in spondyloarthritis-like disease in B10.RIII mice. The development of extra-articular manifestations but absence of arthritis in C57BL/6 mice suggests organ-specific genetic control mechanisms of IL-23 driven inflammation. Discrepancies regarding the phenotype of IL-23 induced disease in different labs and the sexual dimorphism observed in this study warrant further exploration.


2020 ◽  
Author(s):  
Himanshi Singh ◽  
Avinash Y. Gahane ◽  
Virender Singh ◽  
Shreya Ghosh ◽  
Ashwani Kumar Thakur

AbstractBackgroundBiofilm associated infections are the major contributor of mortality, morbidity and financial burden in patients with bacterial infection. Molecules with surfactant behaviour are known to show significant antibiofilm effect against these infections. Thus, newly discovered antibacterial Fmoc-phenylalanine (Fmoc-F) and other Fmoc-amino acids (Fmoc-AA) with surfactant properties, could have potential antibiofilm properties.ObjectivesTo evaluate and characterise the antibiofilm activity of Fmoc-F and some Fmoc-AA against various clinically relevant bacteria.MethodsBiofilm inhibition and eradication was evaluated by crystal violet staining procedure along with scanning electron microscopy (SEM). Attenuated Total Reflection - Fourier Transform Infrared Spectroscopy (ATR-FTIR), Biochemical assays and Congo red staining were employed to investigate mechanism of antibiofilm action.ResultsWe showed that Fmoc-F not only inhibits the biofilm formation in S. aureus and P. aeruginosa, but also eradicates the already formed biofilms over the surface. Further, Fmoc-F coated glass surface resists S. aureus and P. aeruginosa biofilm formation and attachment, when biofilm is grown over the surface. The mechanistic investigation suggests that Fmoc-F reduces the ECM components such as proteins carbohydrates and eDNA in the biofilm and affect its stability via direct interactions with ECM components and/ or indirectly through reducing bacterial cell population. Finally, we showed that Fmoc-F treatment in combination with other antibiotics such as vancomycin and ampicillin synergistically inhibit biofilm formation.ConclusionsOverall, the study demonstrates the potential application of Fmoc-F and other Fmoc-AA molecules individually as well as in combination as antibiofilm agents and antibiofilm coating material for treating biofilm associated infections.


2020 ◽  
Vol 11 ◽  
Author(s):  
Verónica L. Burstein ◽  
Ignacio Beccacece ◽  
Lorena Guasconi ◽  
Cristian J. Mena ◽  
Laura Cervi ◽  
...  

Dermatophytoses (ringworms) are among the most frequent skin infections and are a highly prevalent cause of human disease worldwide. Despite the incidence of these superficial mycoses in healthy people and the compelling evidence on chronic and deep infections in immunocompromised individuals, the mechanisms controlling dermatophyte invasion in the skin are scarcely known. In the last years, the association between certain primary immunodeficiencies and the susceptibility to severe dermatophytosis as well as the evidence provided by novel experimental models mimicking human disease have significantly contributed to deciphering the basic immunological mechanisms against dermatophytes. In this review, we outline the current knowledge on fungal virulence factors involved in the pathogenesis of dermatophytoses and recent evidence from human infections and experimental models that shed light on the cells and molecules involved in the antifungal cutaneous immune response. The latest highlights emphasize the contribution of C-type lectin receptors signaling and the cellular immune response mediated by IL-17 and IFN-γ in the anti-dermatophytic defense and skin inflammation control.


2019 ◽  
Vol 1 (9) ◽  
pp. 3400-3405 ◽  
Author(s):  
S. Tomane ◽  
E. López-Maya ◽  
S. Boujday ◽  
V. Humblot ◽  
J. Marrot ◽  
...  

A reduced polyoxovanadate functionalized with alendronate molecules was used for the synthesis of gold nanoparticles; these hybrid nanomaterials exhibit high antibiofilm activity.


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