scholarly journals The Effect of Oral Probiotic Streptococcus Salivarius K12 on Candida Albicans Biofilm Formation

2020 ◽  
Vol 18 (2) ◽  
Author(s):  
Munirah Mokhtar ◽  
Alia Risma Rismayuddin ◽  
Ridhwan Abdul Wahab ◽  
Muhamad Ashraf Rostam ◽  
Mohd Hamzah Mohd Nasir ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Oral Cancer ◽  
Biofilm Formation ◽  
Streptococcus Salivarius ◽  
Yeast Form ◽  
Cell Numbers ◽  
Crystal Violet Assay ◽  
Common Cancer ◽  
The Media ◽  
Candida Albicans Infection

Introduction:  Oral cancer is the sixth most common cancer worldwide with  Candida albicans  infection being one of the aetiological factors for the disease. Meanwhile,  Streptococcus salivarius  K12 is an oral probiotic that is beneficial to the oral cavity. The objective of the present study is to determine the effect of  S. salivarius  K12 on  C. albicans  biofilm-forming ability with the hypothesis that  S. salivarius  K12 inhibits biofilm formation of  C. albicans  Materials and method: To assess the effect of  S. salivarius  K12 on  C. albicans  biofilm formation,  S. salivarius  K12, lab strain  C. albicans  MYA-4901 and clinical isolates from oral cancer, ALC2 and ALC3 were grown in both nutrient broth (NB) and RPMI. In a mono-species biofilm, 105 of  C. albicans  cells and 106 of  S. salivarius  K12 cells were grown separately in a 96-well plate. In contrast, both microorganisms were combined for polymicrobial biofilms with similar cell numbers as in mono-species. The biofilms were incubated for 72 hours at 37°C and the media were replenished every 24 hours. Finally, the crystal violet assay was conducted, and the optical density was measured at OD620nm.  Results: Polymicrobial biofilms of  C. albicans  (MYA-4901 and ALC3) with  S. salivarius K12 when grown in NB, exhibited a decrease by 64.5 ± 25.8% and 83.7 ± 5.4%, respectively when compared to the expected biofilms which were predominated by yeast form.  Furthermore, polymicrobial biofilms of  C. albicans  (ALC2 and ALC3) with  S. salivarius  K12 showed a decrease by 62.5 ± 25.6% and 55.9 ± 17.1 %, respectively when compared to the expected biofilms when grown in RPMI that were predominated by hyphal form.  Conclusion:  S. salivarius  K12 inhibited polymicrobial biofilms formation of  C. albicans  yeast and hyphal forms, thus supported the hypothesis that  S. salivarius  K12 inhibits biofilm formation of  C. albicans.

scholarly journals Simple Carbohydrate Derivatives Diminish the Formation of Biofilm of the Pathogenic Yeast Candida albicans

Antibiotics ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 10 ◽  
Author(s):  
Olena P. Ishchuk ◽  
Olov Sterner ◽  
Ulf Ellervik ◽  
Sophie Manner
Keyword(s):  
Candida Albicans ◽  
Fungal Pathogen ◽  
Biofilm Formation ◽  
Untreated Control ◽  
Morphological Transition ◽  
Pathogenic Yeast ◽  
Yeast Form ◽  
Human Fungal Pathogen ◽  
Morphological Transitions ◽  
Simple Carbohydrate

The opportunistic human fungal pathogen Candida albicans relies on cell morphological transitions to develop biofilm and invade the host. In the current study, we developed new regulatory molecules, which inhibit the morphological transition of C. albicans from yeast-form cells to cells forming hyphae. These compounds, benzyl α-l-fucopyranoside and benzyl β-d-xylopyranoside, inhibit the hyphae formation and adhesion of C. albicans to a polystyrene surface, resulting in a reduced biofilm formation. The addition of cAMP to cells treated with α-l-fucopyranoside restored the yeast-hyphae switch and the biofilm level to that of the untreated control. In the β-d-xylopyranoside treated cells, the biofilm level was only partially restored by the addition of cAMP, and these cells remained mainly as yeast-form cells.

scholarly journals Streptococcus salivarius K12 inhibits Candida albicans aggregation, biofilm formation and dimorphism

Biofouling ◽  
2021 ◽  
pp. 1-10
Author(s):  
Munirah Mokhtar ◽  
Nurul Alia Risma Rismayuddin ◽  
Aini Syahida Mat Yassim ◽  
Hasna Ahmad ◽  
Ridhwan Abdul Wahab ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Biofilm Formation ◽  
Streptococcus Salivarius

scholarly journals Cajuputs candy impairs Candida albicans and Streptococcus mutans mixed biofilm formation in vitro

F1000Research ◽  
2020 ◽  
Vol 8 ◽  
pp. 1923
Author(s):  
Siska Septiana ◽  
Boy Muchlis Bachtiar ◽  
Nancy Dewi Yuliana ◽  
Christofora Hanny Wijaya
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Candida Albicans ◽  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Microscopy Imaging ◽  
Biofilm Inhibition ◽  
Yeast Form ◽  
Melaleuca Cajuputi ◽  
Scanning Electron

Background: Cajuputs candy (CC), an Indonesian functional food, utilizes the bioactivity of Melaleuca cajuputi essential oil (MCEO) to maintain oral cavity health. Synergistic interaction between Candida albicans and Streptococcus mutans is a crucial step in the pathogenesis of early childhood caries. Our recent study revealed several alternative MCEOs as the main flavors in CC. The capacity of CC to interfere with the fungus-bacterium relationship remains unknown. This study aimed to evaluate CC efficacy to impair biofilm formation by these dual cariogenic microbes. Methods: The inhibition capacity of CC against mixed-biofilm comprising C. albicans and S. mutans was assessed by quantitative (crystal violet assay, tetrazolium salt [MTT] assay, colony forming unit/mL counting, biofilm-related gene expression) and qualitative analysis (light microscopy and scanning electron microscopy). Result: Both biofilm-biomass and viable cells were significantly reduced in the presence of CC. Scanning electron microscopy imaging confirmed this inhibition capacity, demonstrating morphology alteration of C. albicans, along with reduced microcolonies of S. mutans in the biofilm mass. This finding was related to the transcription level of selected biofilm-associated genes, expressed either by C. albicans or S. mutans. Based on qPCR results, CC could interfere with the transition of C. albicans yeast form to the hyphal form, while it suppressed insoluble glucan production by S. mutans. G2 derived from Mojokerto MCEO showed the greatest inhibition activity on the relationship between these cross-kingdom oral microorganisms (p < 0.05). Conclusion: In general, all CC formulas showed biofilm inhibition capacity. Candy derived from Mojokerto MCEO showed the greatest capacity to maintain the yeast form of C. albicans and to inhibit extracellular polysaccharide production by S. mutans. Therefore, the development of dual-species biofilms can be impaired effectively by the CC tested.

scholarly journals A modified crystal violet assay is comparable to XTT reduction assay for quantification of biofilm formation by Candida albicans

2020 ◽  
Author(s):  
Yue Qu ◽  
Shoufeng Yang ◽  
Zhangzhang Chen ◽  
Feifei Su
Keyword(s):  
Candida Albicans ◽  
Crystal Violet ◽  
Biofilm Formation ◽  
Quantitative Methods ◽  
Standard Procedure ◽  
Short Term ◽  
Biofilm Growth ◽  
Human Fungal Pathogen ◽  
Crystal Violet Assay ◽  
Reduction Assay

Abstract Background: The ability of the human fungal pathogen Candida albicans to form biofilms, for example on indwelling medical devices, is a major pathogenic mechanism and has been the focus of intense studies in the fungal pathogenesis field. A key research tool used is the quantitative methods for measuring biofilm formation of C. albicans. Objective: We sought to optimize the conventional crystal violet (CV) staining assay for quantification of biofilm formation by C. albicans and evaluate its performance. Methods: Individual modifications included (i) submerge-washing of microplates to remove non-adherent cells, (ii) heat-fixation, (iii) short-term staining for 3 min, (iv) submerge-washing to remove unbound CV dye, and (v) short-term destaining for 15 min were compared with the standard procedure, and those were superior were incorporated. Performance analysis was carried out for the modified CV assay, in comparison to the conventional CV assay and the XTT [2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide] reduction assay. Results: The modified CV assay demonstrated several advantages in quantitative assessment of biofilm formation of C. albicans over the conventional CV assay, including greater accuracy and reproducibility, shorter experimental time and reduced labor intensity, and was at least comparable to the XTT reduction assay.Conclusion: The modified CV method can be used as an alternative to the XTT reduction assay in quantification of biofilm growth by C. albicans.

scholarly journals Targeting Candida albicans in dual-species biofilms with antifungal treatment reduces Staphylococcus aureus and MRSA in vitro

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0249547
Author(s):  
Yu Luo ◽  
Daniel F. McAuley ◽  
Catherine R. Fulton ◽  
Joana Sá Pessoa ◽  
Ronan McMullan ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Candida Albicans ◽  
Bacterial Cell ◽  
Biofilm Formation ◽  
Liposomal Amphotericin ◽  
Antifungal Treatment ◽  
Resistant Bacteria ◽  
Respiratory Pathogens ◽  
Cell Numbers

Polymicrobial biofilms consisting of fungi and bacteria are frequently formed on endotracheal tubes and may contribute to development of ventilator associated pneumonia (VAP) in critically ill patients. This study aimed to determine the role of early Candida albicans biofilms in supporting dual-species (dual-kingdom) biofilm formation with respiratory pathogens in vitro, and investigated the effect of targeted antifungal treatment on bacterial cells within the biofilms. Dual-species biofilm formation between C. albicans and three respiratory pathogens commonly associated with VAP (Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus) was studied using quantitative PCR. It was shown that early C. albicans biofilms enhanced the numbers of E. coli and S. aureus (including methicillin resistant S. aureus; MRSA) but not P. aeruginosa within dual-species biofilms. Transwell assays demonstrated that contact with C. albicans was required for the increased bacterial cell numbers observed. Total Internal Reflection Fluorescence microscopy showed that both wild type and hyphal-deficient C. albicans provided a scaffold for initial bacterial adhesion in dual species biofilms. qPCR results suggested that further maturation of the dual-species biofilm significantly increased bacterial cell numbers, except in the case of E.coli with hyphal-deficient C. albicans (Ca_gcn5Δ/Δ). A targeted preventative approach with liposomal amphotericin (AmBisome®) resulted in significantly decreased numbers of S. aureus in dual-species biofilms, as determined by propidium monoazide-modified qPCR. Similar results were observed when dual-species biofilms consisting of clinical isolates of C. albicans and MRSA were treated with liposomal amphotericin. However, reductions in E. coli numbers were not observed following liposomal amphotericin treatment. We conclude that early C. albicans biofilms have a key supporting role in dual-species biofilms by enhancing bacterial cell numbers during biofilm maturation. In the setting of increasing antibiotic resistance, an important and unexpected consequence of antifungal treatment of dual-species biofilms, is the additional benefit of decreased growth of multi-drug resistant bacteria such as MRSA, which could represent a novel future preventive strategy.

scholarly journals Inhibition of Distinct Proline- or N-Acetylglucosamine-Induced Hyphal Formation Pathways by Proline Analogs in Candida albicans

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Tatsuki Sato ◽  
Hisashi Hoshida ◽  
Rinji Akada
Keyword(s):  
Candida Albicans ◽  
Biofilm Formation ◽  
Culture Medium ◽  
Turning Point ◽  
Fetal Bovine Serum ◽  
Yeast Form ◽  
Induction Condition ◽  
Fetal Bovine ◽  
Virulence Property ◽  
Hyphal Formation

Candida albicans undergoes a yeast-to-hyphal transition that has been recognized as a virulence property as well as a turning point leading to biofilm formation associated with candidiasis. It is known that yeast-to-hyphal transition is induced under complex environmental conditions including temperature (above 35°C), pH (greater than 6.5), CO2, N-acetylglucosamine (GlcNAc), amino acids, RPMI-1640 synthetic culture medium, and blood serum. To identify the hyphal induction factor in the RPMI-1640 medium, we examined each component of RPMI-1640 and established a simple hyphal induction condition, that is, incubation in L-proline solution at 37°C. Incubation in GlcNAc solution alone, which is not contained in RPMI-1640, without any other materials was also identified as another simple hyphal induction condition. To inhibit hyphal formation, proline and GlcNAc analogs were examined. Among the proline analogs used, L-azetidine-2-carboxylic acid (AZC) inhibited hyphal induction under both induction conditions, but L-4-thiazolidinecarboxylic acid (T4C) specifically inhibited proline-induced hyphal formation only, while α-N-methyl-L-proline (mPro) selectively inhibited GlcNAc-induced hyphal formation. Hyphal formation in fetal bovine serum was also inhibited by AZC or T4C together with mPro without affecting the proliferation of yeast form. These results indicate that these proline analogs are ideal inhibitors of yeast-to-hyphal transition in C. albicans.

scholarly journals Simple Carbohydrate Derivatives Diminish the Formation of Biofilm of Candida albicans

2019 ◽  
Author(s):  
Olena P. Ishchuk ◽  
Olov Sterner ◽  
Ulf Ellervik ◽  
Sophie Manner
Keyword(s):  
Candida Albicans ◽  
Fungal Pathogen ◽  
Biofilm Formation ◽  
Untreated Control ◽  
Morphological Transition ◽  
Yeast Form ◽  
Polystyrene Surface ◽  
Human Fungal Pathogen ◽  
Morphological Transitions ◽  
Simple Carbohydrate

Abstract The opportunistic human fungal pathogen Candida albicans rely on cell morphological transitions to develop biofilm and invade the host. In the current study, we developed new regulatory molecules, which inhibit the morphological transition of C. albicans from yeast-form cells to cells forming hyphae. These compounds, benzyl α-L-fucopyranoside and benzyl β-D-xylopyranoside, inhibit the morphological switching and adhesion of C. albicans to a polystyrene surface, resulting in a reduced biofilm formation. The addition of cAMP to cells treated with α-L-fucopyranoside restored the yeast-hyphae switch and the biofilm level to that of the untreated control. In the β-D-xylopyranoside treated cells, the biofilm level was only partially restored by the addition of cAMP, and these cells remained mainly as yeast-form cells.

scholarly journals Simple Carbohydrate Derivatives Diminish the Formation of Biofilm of Candida albicans

2019 ◽  
Author(s):  
Olena P. Ishchuk ◽  
Olov Sterner ◽  
Ulf Ellervik ◽  
Sophie Manner
Keyword(s):  
Candida Albicans ◽  
Fungal Pathogen ◽  
Biofilm Formation ◽  
Untreated Control ◽  
Morphological Transition ◽  
Yeast Form ◽  
Polystyrene Surface ◽  
Human Fungal Pathogen ◽  
Morphological Transitions ◽  
Simple Carbohydrate

Abstract The opportunistic human fungal pathogen Candida albicans rely on cell morphological transitions to develop biofilm and invade the host. In the current study, we developed new regulatory molecules, which inhibit the morphological transition of C. albicans from yeast-form cells to cells forming hyphae. These compounds, benzyl α-L-fucopyranoside and benzyl β-D-xylopyranoside, inhibit the morphological switching and adhesion of C. albicans to a polystyrene surface, resulting in a reduced biofilm formation. The addition of cAMP to cells treated with α-L-fucopyranoside restored the yeast-hyphae switch and the biofilm level to that of the untreated control. In the β-D-xylopyranoside treated cells, the biofilm level was only partially restored by the addition of cAMP, and these cells remained mainly as yeast-form cells.

scholarly journals Candida albicans adhesin Als3p is dispensable for virulence in the mouse model of disseminated candidiasis

Microbiology ◽  
2011 ◽  
Vol 157 (6) ◽  
pp. 1806-1815 ◽  
Author(s):  
Ian A. Cleary ◽  
Sara M. Reinhard ◽  
C. Lindsay Miller ◽  
Craig Murdoch ◽  
Martin H. Thornhill ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Candida Albicans ◽  
Murine Model ◽  
Biofilm Formation ◽  
Microvascular Endothelial Cells ◽  
Wild Type ◽  
Yeast Form ◽  
Disseminated Candidiasis ◽  
Microvascular Endothelial ◽  
Null Strain

The presence of specific proteins, including Ece1p, Hwp1p and Als3p, distinguishes the Candida albicans hyphal cell wall from that of yeast-form cells. These proteins are thought to be important for the ability of C. albicans cells to adhere to living and non-living surfaces and for the cell-to-cell adhesion necessary for biofilm formation, and also to be pivotal in mediating C. albicans interactions with endothelial cells. Using an in vitro flow adhesion assay, we previously observed that yeast cells bind in greater numbers to human microvascular endothelial cells than do hyphal or pseudohyphal cells. This is consistent with previous observations that, in a murine model of disseminated candidiasis, cells locked in the yeast form can efficiently escape the bloodstream and invade host tissues. To more precisely explore the role of Als3p in adhesion and virulence, we deleted both copies of ALS3 in a wild-type C. albicans strain. In agreement with previous studies, our als3Δ null strain formed hyphae normally but was defective in biofilm formation. Whilst ALS3 was not expressed in our null strain, hypha-specific genes such as ECE1 and HWP1 were still induced appropriately. Both the yeast form and the hyphal form of the als3Δ strain adhered to microvascular endothelial cells to the same extent as a wild-type strain under conditions of flow, indicating that Als3p is not a significant mediator of the initial interaction between fungal cells and the endothelium. Finally, in a murine model of haematogenously disseminated candidiasis the mutant als3Δ remained as virulent as the wild-type parent strain.

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