Antibiosis interaction of Staphylococccus aureus on Aspergillus fumigatus assessed in vitro by mixed biofilm formation

Ramírez-Granillo, Adrián; Canales, Maria Gabriela Medina; Espindola, M. E. Sanchez; Rivera, María Angeles Martínez; Lucio, Victor Manuel Bautista de; Tovar, Aída Verónica Rodríguez

doi:10.1186/s12866-015-0363-2

Cited by 60 publications

(53 citation statements)

References 43 publications

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“…The hyphae are highly ramified and the cell wall is thicker in the presence of the bacteria (Fig 5). These findings are entirely opposite to the results of Ramirez Granillo et al [35] where the fungal structures forming the mixed biofilm of A . fumigatus and S .…”

Section: Discussioncontrasting

confidence: 99%

“…fumigatus or S . maltophilia and other microorganisms [15, 32–35], but never between these two microorganisms. The originality of this study was to develop for the first time an in vitro model of mixed biofilm of A .…”

Section: Discussionmentioning

confidence: 99%

“…aeruginosa [16, 34] or A . fumigatus and Staphylococcus aureus [35]. An example of cellular structures that can support such interactions is cellulose, which is synthesized by several bacterial species and has a critical role in bacterial attachment to fungi, and in particular to chitin.…”

Section: Discussionmentioning

confidence: 99%

“…aeruginosa [15] or S . aureus [35]. Moreover, CLSM showed a difference of the hyphae phenotype between the single fungal and mixed biofilm (Fig 4).…”

Section: Discussionmentioning

confidence: 99%

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Characteristics of Aspergillus fumigatus in Association with Stenotrophomonas maltophilia in an In Vitro Model of Mixed Biofilm

et al. 2016

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BackgroundBiofilms are communal structures of microorganisms that have long been associated with a variety of persistent infections poorly responding to conventional antibiotic or antifungal therapy. Aspergillus fumigatus fungus and Stenotrophomonas maltophilia bacteria are examples of the microorganisms that can coexist to form a biofilm especially in the respiratory tract of immunocompromised patients or cystic fibrosis patients. The aim of the present study was to develop and assess an in vitro model of a mixed biofilm associating S. maltophilia and A. fumigatus by using analytical and quantitative approaches.Materials and MethodsAn A. fumigatus strain (ATCC 13073) expressing a Green Fluorescent Protein (GFP) and an S. maltophilia strain (ATCC 13637) were used. Fungal and bacterial inocula (105 conidia/mL and 106 cells/mL, respectively) were simultaneously deposited to initiate the development of an in vitro mixed biofilm on polystyrene supports at 37°C for 24 h. The structure of the biofilm was analysed via qualitative microscopic techniques like scanning electron and transmission electron microscopy, and fluorescence microscopy, and by quantitative techniques including qPCR and crystal violet staining.ResultsAnalytic methods revealed typical structures of biofilm with production of an extracellular matrix (ECM) enclosing fungal hyphae and bacteria. Quantitative methods showed a decrease of A. fumigatus growth and ECM production in the mixed biofilm with antibiosis effect of the bacteria on the fungi seen as abortive hyphae, limited hyphal growth, fewer conidia, and thicker fungal cell walls.ConclusionFor the first time, a mixed A. fumigatus—S. maltophilia biofilm was validated by various analytical and quantitative approaches and the bacterial antibiosis effect on the fungus was demonstrated. The mixed biofilm model is an interesting experimentation field to evaluate efficiency of antimicrobial agents and to analyse the interactions between the biofilm and the airways epithelium.

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Section: Discussioncontrasting

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…aeruginosa [15] or S . aureus [35]. Moreover, CLSM showed a difference of the hyphae phenotype between the single fungal and mixed biofilm (Fig 4).…”

Section: Discussionmentioning

confidence: 99%

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Characteristics of Aspergillus fumigatus in Association with Stenotrophomonas maltophilia in an In Vitro Model of Mixed Biofilm

et al. 2016

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“…In general, these studies mainly involve the biofilm maturity stage and the chemical composition of the ECM, with few images of the biofilm stages, but any one of these described all of the stages of biofilm formation. Thus, the information is different from the contributions made by our working group [ 5 , 8 , 11 , 12 , 17 , 18 , 23 – 28 ].…”

Section: Discussionmentioning

confidence: 74%

Analysis and description of the stages of Aspergillus fumigatus biofilm formation using scanning electron microscopy

et al. 2016

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BackgroundBiofilms are a highly structured consortia of microorganisms that adhere to a substrate and are encased within an extracellular matrix (ECM) that is produced by the organisms themselves. Aspergillus fumigatus is a biotechnological fungus that has a medical and phytopathogenic significance, and its biofilm occurs in both natural and artificial environments; therefore, studies on the stages observed in biofilm formation are of great significance due to the limited knowledge that exists on this specific topic and because there are multiple applications that are being carried out.ResultsGrowth curves were obtained from the soil and clinical isolates of the A. fumigatus biofilm formation. The optimal conditions for both of the isolates were inocula of 1 × 106 conidia/mL, incubated at 28 °C during 24 h; these showed stages similar to those described in classic microbial growth: the lag, exponential, and stationary phases. However, the biofilms formed at 37 °C were uneven.The A. fumigatus biofilm was similar regardless of the isolation source, but differences were presented according to the incubation temperature. The biofilm stages included the following: 1) adhesion to the plate surface (4 h), cell co-aggregation and exopolymeric substance (EPS) production; 2) conidial germination into hyphae (8-12 h), development, hyphal elongation, and expansion with channel formation (16-20 h); and 3) biofilm maturation as follows: mycelia development, hyphal layering networks, and channels formation, and high structural arrangement of the mycelia that included hyphal anastomosis and an extensive production of ECM (24 h); the ECM covered, surrounded and strengthened the mycelial arrangements, particular at 37 °C. In the clinical isolate, irregular fungal structures, such as microhyphae that are short and slender hyphae, occurred; 4) In cell dispersion, the soil isolate exhibited higher conidia than the clinical isolate, which had the capacity to germinate and generate new mycelia growth (24 h). In addition, we present images on the biofilm’s structural arrangement and chemical composition using fluorochromes to detect metabolic activity (FUNI) and mark molecules, such as chitin, DNA, mannose, glucose and proteins.ConclusionsTo our knowledge, this is the first time that, in vitro, scanning electronic microscopy (SEM) images of the stages of A. fumigatus biofilm formation have been presented with a particular emphasis on the high hyphal organization and in diverse ECM to observe biofilm maturation.

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Structural evaluation of polymeric microbial films grown on kefir loaded with Maytenus rigida extract

Silva

Matos

Estevam

et al. 2020

Microscopy Res & Technique

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Kefir is a probiotic that has several health promising properties. Its grains can form microbial films on different types of substrates. In the present work, the surface characteristics of kefir biofilms associated with Maytenus rigida Mart. extract were minutely studied. Three different concentrations of plant extract were included in the biofilm forming solutions, where fresh grains of kefir were inoculated. The results showed that the plant extract was successfully incorporated into the exopolysaccharide matrix of the biofilm. The main chemical components found linked to the plant extract were triterpenes. The crystallinity of biofilms increased with the addition of the plant extract. The morphology revealed that at low concentrations of the extract there was a prevalence of lactobacilli, while at high concentrations yeasts were more observed. Adhesion and wettability were higher for biofilm with less extract. These results revealed that a combination of plant extract and kefir's exopolysaccharide could form biofilms with chemical and topographic properties of great interest in regenerative medicine.

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Antibiosis interaction of Staphylococccus aureus on Aspergillus fumigatus assessed in vitro by mixed biofilm formation

Cited by 60 publications

References 43 publications

Characteristics of Aspergillus fumigatus in Association with Stenotrophomonas maltophilia in an In Vitro Model of Mixed Biofilm

Characteristics of Aspergillus fumigatus in Association with Stenotrophomonas maltophilia in an In Vitro Model of Mixed Biofilm

Analysis and description of the stages of Aspergillus fumigatus biofilm formation using scanning electron microscopy

Structural evaluation of polymeric microbial films grown on kefir loaded with Maytenus rigida extract

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