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The purpose of this research was to determine Aspergillus fumigatus conidial viability and its biofilm formation upon treatment with triclosan and amphotericin-B loaded liposomes. A. fumigatus was treated with the antimicrobials, triclosan and liposomal amphotericin-B (L-AMB), in single and combined supplementation. To quantify the cells’ viability upon treatments, resazurin-based viability assay was performed. Confocal laser scanning microscopy was done by applying FUN-1 stain to screen the role of the agents on extracellular polymeric substances. Total A. fumigatus biomass upon treatments was estimated by using crystal violet-based assay. To study the agents’ effect on the conidial viability, flow cytometry analysis was performed. Expression levels of A. fumigatus genes encoding cell wall proteins, α-(1,3)-glucans and galactosaminogalactan were analysed by real-time polymerase chain reaction assay. A synergistic interaction occurred between triclosan and L-AMB when they were added sequentially (triclosan+L-AMB) at their sub-minimum inhibitory concentrations, the triclosan and L-AMB MICs were dropped to 0.6 and 0.2 mg/L, respectively, from 2 and 1 mg/L. Besides, L-AMB and triclosan contributed to the down-regulation of α-(1,3)-glucan and galactosaminogalactan in A. fumigatus conidia and resulted in less conidia aggregation and mycelia adhesion to the biotic/abiotic surfaces; A. fumigatus conidia -became hydrophilic upon treatment, as a result of rodlet layer being masked by a hydrophilic layer or modified by the ionic strength of the rodlet layer. In A. fumigatus, the potential mechanisms of action for L-AMB might be through killing the cells and for triclosan through interrupting the cells’ development as a consequence of quorum quenching.
Keywords
Conidia, biofilm, combination treatment, flow cytometry, quorum quenching
Figure 1
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Figure 3
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Figure 5
This is a list of supplementary files associated with this preprint. Click to download.
- Fig.1.png
A. fumigatus inoculum was cultured and treated with triclosan doses in RPMI1640 medium at 37°C under static condition. Effective dose of triclosan against A. fumigatus was determined by resazurine- based viability assay. Its MIC was calculated as 2 mg/L, which inhibits the viability by ~ 72% after 24 h of incubation. *P ≤ 0.05, **P ≤ 0.01 and ***P ≤ 0.001 (SEM bars are shown for n = 3)
- Fig.2.png
A. fumigatus inoculum was cultured and treated with L-AMB doses in RPMI-1640 medium at 37°C under static condition. Effective dose of L-AMB against A. fumigatus was determined by resazurine- based viability assay. Its MIC was calculated as 1 mg/L, which inhibits the viability by 68% after 24 h of incubation. *P ≤ 0.05 and **P ≤ 0.01 and ***P ≤ 0.001 (SEM bars are shown for n = 3)
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This preprint is under consideration at World Journal of Microbiology and Biotechnology. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research Article
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Review
Version 1
Posted 18 Nov, 2021
No community comments so far
Reviewers invited
Invitations sent on 16 Nov, 2021
Editor invited
On 05 Nov, 2021
Editor assigned
On 02 Nov, 2021
First submitted
On 01 Nov, 2021
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
The purpose of this research was to determine Aspergillus fumigatus conidial viability and its biofilm formation upon treatment with triclosan and amphotericin-B loaded liposomes. A. fumigatus was treated with the antimicrobials, triclosan and liposomal amphotericin-B (L-AMB), in single and combined supplementation. To quantify the cells’ viability upon treatments, resazurin-based viability assay was performed. Confocal laser scanning microscopy was done by applying FUN-1 stain to screen the role of the agents on extracellular polymeric substances. Total A. fumigatus biomass upon treatments was estimated by using crystal violet-based assay. To study the agents’ effect on the conidial viability, flow cytometry analysis was performed. Expression levels of A. fumigatus genes encoding cell wall proteins, α-(1,3)-glucans and galactosaminogalactan were analysed by real-time polymerase chain reaction assay. A synergistic interaction occurred between triclosan and L-AMB when they were added sequentially (triclosan+L-AMB) at their sub-minimum inhibitory concentrations, the triclosan and L-AMB MICs were dropped to 0.6 and 0.2 mg/L, respectively, from 2 and 1 mg/L. Besides, L-AMB and triclosan contributed to the down-regulation of α-(1,3)-glucan and galactosaminogalactan in A. fumigatus conidia and resulted in less conidia aggregation and mycelia adhesion to the biotic/abiotic surfaces; A. fumigatus conidia -became hydrophilic upon treatment, as a result of rodlet layer being masked by a hydrophilic layer or modified by the ionic strength of the rodlet layer. In A. fumigatus, the potential mechanisms of action for L-AMB might be through killing the cells and for triclosan through interrupting the cells’ development as a consequence of quorum quenching.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
This is a list of supplementary files associated with this preprint. Click to download.
- Fig.1.png
A. fumigatus inoculum was cultured and treated with triclosan doses in RPMI1640 medium at 37°C under static condition. Effective dose of triclosan against A. fumigatus was determined by resazurine- based viability assay. Its MIC was calculated as 2 mg/L, which inhibits the viability by ~ 72% after 24 h of incubation. *P ≤ 0.05, **P ≤ 0.01 and ***P ≤ 0.001 (SEM bars are shown for n = 3)
- Fig.2.png
A. fumigatus inoculum was cultured and treated with L-AMB doses in RPMI-1640 medium at 37°C under static condition. Effective dose of L-AMB against A. fumigatus was determined by resazurine- based viability assay. Its MIC was calculated as 1 mg/L, which inhibits the viability by 68% after 24 h of incubation. *P ≤ 0.05 and **P ≤ 0.01 and ***P ≤ 0.001 (SEM bars are shown for n = 3)
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