scholarly journals The Role of Candida albicans Virulence Factors in the Formation of Multispecies Biofilms With Bacterial Periodontal Pathogens

2022 ◽  
Vol 11 ◽  
Author(s):  
Dorota Satala ◽  
Miriam Gonzalez-Gonzalez ◽  
Magdalena Smolarz ◽  
Magdalena Surowiec ◽  
Kamila Kulig ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Cell Surface ◽  
Oral Cavity ◽  
Virulence Factors ◽  
Bacterial Species ◽  
Complex Structure ◽  
Diagnostic Methods ◽  
Fungal Virulence ◽  
Complex Species ◽  
Periodontal Tissues

Periodontal disease depends on the presence of different microorganisms in the oral cavity that during the colonization of periodontal tissues form a multispecies biofilm community, thus allowing them to survive under adverse conditions or facilitate further colonization of host tissues. Not only numerous bacterial species participate in the development of biofilm complex structure but also fungi, especially Candida albicans, that often commensally inhabits the oral cavity. C. albicans employs an extensive armory of various virulence factors supporting its coexistence with bacteria resulting in successful host colonization and propagation of infection. In this article, we highlight various aspects of individual fungal virulence factors that may facilitate the collaboration with the associated bacterial representatives of the early colonizers of the oral cavity, the bridging species, and the late colonizers directly involved in the development of periodontitis, including the “red complex” species. In particular, we discuss the involvement of candidal cell surface proteins—typical fungal adhesins as well as originally cytosolic “moonlighting” proteins that perform a new function on the cell surface and are also present within the biofilm structures. Another group of virulence factors considered includes secreted aspartic proteases (Sap) and other secreted hydrolytic enzymes. The specific structure of the candidal cell wall, dynamically changing during morphological transitions of the fungus that favor the biofilm formation, is equally important and discussed. The non-protein biofilm-composing factors also show dynamic variability upon the contact with bacteria, and their biosynthesis processes could be involved in the stability of mixed biofilms. Biofilm-associated changes in the microbe communication system using different quorum sensing molecules of both fungal and bacterial cells are also emphasized in this review. All discussed virulence factors involved in the formation of mixed biofilm pose new challenges and influence the successful design of new diagnostic methods and the application of appropriate therapies in periodontal diseases.

scholarly journals Nitrosative Stress Biomarkers in the Non-Stimulated and Stimulated Saliva, as well as Gingival Crevicular Fluid of Patients with Periodontitis: Review and Clinical Study

Antioxidants ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 259 ◽  
Author(s):  
Joanna Toczewska ◽  
Tomasz Konopka ◽  
Anna Zalewska ◽  
Mateusz Maciejczyk
Keyword(s):  
Oral Cavity ◽  
Nitrosative Stress ◽  
Gingival Crevicular Fluid ◽  
Disease Process ◽  
Primary Source ◽  
Diagnostic Methods ◽  
Periodontal Tissues ◽  
Crevicular Fluid ◽  
The Impact ◽  
Gingival Fluid

Diagnosis of periodontopathy is complex and includes defining the cause, type, stage, and grade of periodontitis. Therefore, alternative diagnostic methods are sought to indicate the progression of inflammation or to determine the effectiveness of therapy. Gingival crevicular fluid (GCF) biomarkers can be particularly useful because they most likely reflect the disease process of the periodontal tissues. However, the difficulty of collecting GCF for testing is the reason for the limited use in diagnostics. Because periodontitis is the primary source of nitrogen free radicals in the oral cavity, the aim of the study was to evaluate the biomarkers of nitrosative stress (nitric oxide, peroxynitrite, and S-nitrosothiols) in GCF, non-stimulated and stimulated saliva of 90 patients with periodontitis. The study group was divided into two subgroups, depending on the stage of the disease severity. We showed a significantly higher concentration of all assessed biomarkers in the non-stimulated and stimulated saliva of patients with periodontitis. However, significant changes in GCF has been shown only for peroxynitrite. The studied biomarkers did not correlate with clinical periodontal status, which probably results from their short-duration activity and the impact on a few factors in the oral cavity. Saliva and gingival fluid are not very useful in the differential diagnosis of periodontitis.

scholarly journals Oestrogen promotes innate immune evasion of Candida albicans through inactivation of the alternative complement system

2020 ◽  
Author(s):  
Pizga Kumwenda ◽  
Fabien Cottier ◽  
Ben Keevan ◽  
Hannah Gallagher ◽  
Hung-Ji Tsai ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Cell Surface ◽  
Immune Evasion ◽  
Fungal Infections ◽  
Hormone Replacement ◽  
Factor H ◽  
Innate Immune ◽  
Fungal Cell ◽  
Fungal Virulence ◽  
Innate Immune Evasion

AbstractGender is a risk factor for several infections that, for many pathogens, has been linked to sex hormones impacting host immunity and directly affecting microbial virulence. Candida albicans is a commensal of the urogenital tract and the predominant cause of vulvovaginal candidiasis (VVC). Factors that increase circulatory oestrogen levels like pregnancy, the use of oral contraceptives, and hormone replacement therapy predispose women to VVC, but the reasons for this are largely unknown. Here, we investigate how adaptation of C. albicans to oestrogen impacts the fungal host-pathogen interaction. Physiologically relevant concentrations of oestrogen promoted fungal virulence by enabling C. albicans to avoid the actions of the innate immune system. Oestrogen-induced innate immune evasion was mediated via inhibition of opsonophagocytosis through enhanced acquisition of the human complement regulatory protein, Factor H, on the fungal cell surface. Oestrogen induced accumulation of Factor H was dependent on the fungal cell surface protein Gpd2, with oestrogen dependent derepression of GPD2 being mediated via a non-canonical signalling pathway involving Ebp1 and Bcr1. Therefore, we propose that, in addition to affecting the antifungal potential of vaginal epithelial cells, elevated oestrogen levels predispose women to VVC by directly enhancing fungal pathogenicity through the inactivation of complement. The discovery of this new hormone sensing pathway might pave the way in explaining gender biases associated with fungal infections and may provide an alternative approach to improving women’s health.

scholarly journals Dramatic Improvement of CRISPR/Cas9 Editing in Candida albicans by Increased Single Guide RNA Expression

mSphere ◽  
2017 ◽  
Vol 2 (2) ◽  
Author(s):  
Henry Ng ◽  
Neta Dean
Keyword(s):  
Candida Albicans ◽  
Genome Editing ◽  
Virulence Factors ◽  
Fungal Pathogen ◽  
Fungal Virulence ◽  
Important Conclusion ◽  
Content Type ◽  
Guide Rna ◽  
Improve Efficiency ◽  
Human Fungal Pathogen

ABSTRACT Candida albicans is an important human fungal pathogen. An understanding of fungal virulence factors has been slow because C. albicans is genetically intractable. The recent development of CRISPR/Cas in C. albicans (V. K. Vyas, M. I. Barrasa, G. R. Fink, Sci Adv 1:e1500248, 2015, https://doi.org/10.1126/sciadv.1500248 ) has the potential to circumvent this problem. However, as has been found in other organisms, CRISPR/Cas mutagenesis efficiency can be frustratingly variable. Here, we systematically examined parameters hypothesized to alter sgRNA intracellular levels in order to optimize CRISPR/Cas in C. albicans. Our most important conclusion is that increased sgRNA expression and maturation dramatically improve efficiency of CRISPR/Cas mutagenesis in C. albicans by ~10-fold. Thus, we anticipate that the modifications described here will further advance the application of CRISPR/Cas for genome editing in C. albicans. The clustered regularly interspaced short palindromic repeat system with CRISPR-associated protein 9 nuclease (CRISPR/Cas9) has emerged as a versatile tool for genome editing in Candida albicans. Mounting evidence from other model systems suggests that the intracellular levels of single guide RNA (sgRNA) limit the efficiency of Cas9-dependent DNA cleavage. Here, we tested this idea and describe a new means of sgRNA delivery that improves previously described methods by ~10-fold. The efficiency of Cas9/sgRNA-dependent cleavage and repair of a single-copy yeast enhanced monomeric red fluorescent protein (RFP) gene was measured as a function of various parameters that are hypothesized to affect sgRNA accumulation, including transcriptional and posttranscriptional processing. We analyzed different promoters (SNR52, ADH1, and tRNA), as well as different posttranscriptional RNA processing schemes that serve to generate or stabilize mature sgRNA with precise 5′ and 3′ ends. We compared the effects of flanking sgRNA with self-cleaving ribozymes or by tRNA, which is processed by endogenous RNases. These studies demonstrated that sgRNA flanked by a 5′ tRNA and transcribed by a strong RNA polymerase II ADH1 promoter increased Cas9-dependent RFP mutations by 10-fold. Examination of double-strand-break (DSB) repair in strains hemizygous for RFP demonstrated that both homology-directed and nonhomologous end-joining pathways were used to repair breaks. Together, these results support the model that gRNA expression can be rate limiting for efficient CRISPR/Cas mutagenesis in C. albicans. IMPORTANCE Candida albicans is an important human fungal pathogen. An understanding of fungal virulence factors has been slow because C. albicans is genetically intractable. The recent development of CRISPR/Cas in C. albicans (V. K. Vyas, M. I. Barrasa, G. R. Fink, Sci Adv 1:e1500248, 2015, https://doi.org/10.1126/sciadv.1500248 ) has the potential to circumvent this problem. However, as has been found in other organisms, CRISPR/Cas mutagenesis efficiency can be frustratingly variable. Here, we systematically examined parameters hypothesized to alter sgRNA intracellular levels in order to optimize CRISPR/Cas in C. albicans. Our most important conclusion is that increased sgRNA expression and maturation dramatically improve efficiency of CRISPR/Cas mutagenesis in C. albicans by ~10-fold. Thus, we anticipate that the modifications described here will further advance the application of CRISPR/Cas for genome editing in C. albicans.

scholarly journals Candida species distribution, genotyping and virulence factors of Candida albicans isolated from the oral cavity of kidney transplant recipients of two geographic regions of Brazil

2014 ◽  
Vol 14 (1) ◽  
Author(s):  
Walicyranison Plinio da Silva-Rocha ◽  
Vitor Luiz de Brito Lemos ◽  
Terezinha Inês Estivalet Svidizisnki ◽  
Eveline Pipolo Milan ◽  
Guilherme Maranhão Chaves
Keyword(s):  
Candida Albicans ◽  
Oral Cavity ◽  
Kidney Transplant ◽  
Virulence Factors ◽  
Species Distribution ◽  
Candida Species ◽  
Transplant Recipients ◽  
Kidney Transplant Recipients ◽  
Geographic Regions

scholarly journals Fungi pathogenic to humans: molecular bases of virulence of Candida albicans, Cryptococcus neoformans and Aspergillus fumigatus.

2009 ◽  
Vol 56 (2) ◽  
Author(s):  
Justyna Karkowska-Kuleta ◽  
Maria Rapala-Kozik ◽  
Andrzej Kozik
Keyword(s):  
Candida Albicans ◽  
Aspergillus Fumigatus ◽  
Cryptococcus Neoformans ◽  
Virulence Factors ◽  
Immunosuppressive Agents ◽  
Hydrolytic Enzymes ◽  
Host Cells ◽  
Phenotypic Switching ◽  
Human Organism ◽  
Fungal Virulence

The frequency of severe systemic fungal diseases has increased in the last few decades. The clinical use of antibacterial drugs, immunosuppressive agents after organ transplantation, cancer chemotherapy, and advances in surgery are associated with increasing risk of fungal infections. Opportunistic pathogens from the genera Candida and Aspergillus as well as pathogenic fungi from the genus Cryptococcus can invade human organism and may lead to mucosal and skin infections or to deep-seated mycoses of almost all inner organs, especially in immunocompromised patients. Nowadays, there are some effective antifungal agents, but, unfortunately, some of the pathogenic species show increasing resistance. The identification of fungal virulence factors and recognition of mechanisms of pathogenesis may lead to development of new efficient antifungal therapies. This review is focused on major virulence factors of the most common fungal pathogens of humans: Candida albicans, Aspergillus fumigatus and Cryptococcus neoformans. The adherence to host cells and tissues, secretion of hydrolytic enzymes, phenotypic switching and morphological dimorphism contribute to C. albicans virulence. The ability to grow at 37 degrees C, capsule synthesis and melanin formation are important virulence factors of C. neoformans. The putative virulence factors of A. fumigatus include production of pigments, adhesion molecules present on the cell surface and secretion of hydrolytic enzymes and toxins.

Cold atmospheric plasma inhibits the growth of Candida albicans by affecting ergosterol biosynthesis and suppresses the fungal virulence factors in vitro

2016 ◽  
Vol 13 ◽  
pp. 66-72 ◽  
Author(s):  
Neda Rahimi-Verki ◽  
Atena Shapoorzadeh ◽  
Mehdi Razzaghi-Abyaneh ◽  
Seyed-Mohammad Atyabi ◽  
Masoomeh Shams-Ghahfarokhi ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Virulence Factors ◽  
Atmospheric Plasma ◽  
Ergosterol Biosynthesis ◽  
Fungal Virulence ◽  
Cold Atmospheric Plasma

scholarly journals New Cell Surface Protein Involved in Biofilm Formation by Streptococcus parasanguinis

2011 ◽  
Vol 79 (8) ◽  
pp. 3239-3248 ◽  
Author(s):  
Xiaobo Liang ◽  
Yi-Ywan M. Chen ◽  
Teresa Ruiz ◽  
Hui Wu
Keyword(s):  
Cell Surface ◽  
Oral Cavity ◽  
Biofilm Formation ◽  
Bacterial Species ◽  
Surface Protein ◽  
Cell Surface Protein ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
Dental Biofilm ◽  
Streptococcus Parasanguinis

ABSTRACTDental biofilm formation is critical for maintaining the healthy microbial ecology of the oral cavity. Streptococci are predominant bacterial species in the oral cavity and play important roles in the initiation of plaque formation. In this study, we identified a new cell surface protein, BapA1, fromStreptococcus parasanguinisFW213 and determined that BapA1 is critical for biofilm formation. Sequence analysis revealed that BapA1 possesses a typical cell wall-sorting signal for cell surface-anchored proteins from Gram-positive bacteria. No functional orthologue was reported in other streptococci. BapA1 possesses nine putative pilin isopeptide linker domains which are crucial for pilus assembly in a number of Gram-positive bacteria. Deletion of the 3′ portion ofbapA1generated a mutant that lacks surface-anchored BapA1 and abolishes formation of short fibrils on the cell surface. The mutant failed to form biofilms and exhibited reduced adherence to anin vitrotooth model. The BapA1 deficiency also inhibited bacterial autoaggregation. The N-terminal muramidase-released-protein-like domain mediated BapA1-BapA1 interactions, suggesting that BapA1-mediated cell-cell interactions are important for bacterial autoaggregation and biofilm formation. Furthermore, the BapA1-mediated bacterial adhesion and biofilm formation are independent of a fimbria-associated serine-rich repeat adhesin, Fap1, demonstrating that BapA1 is a new streptococcal adhesin.

scholarly journals Periodontopathogens Detection in White-Cheeked Spider Monkey (Ateles marginatus)

2018 ◽  
Vol 46 ◽  
pp. 4
Author(s):  
Andresa De Cássia Martini ◽  
Lianna Ghisi Gomes ◽  
Fernanda Harumi Mruyama ◽  
Luciana Maria Curtio Soares ◽  
Sandra Helena Ramiro Corrêa ◽  
...  
Keyword(s):  
Oral Cavity ◽  
Periodontal Disease ◽  
Bacterial Species ◽  
Disease Stage ◽  
Average Weight ◽  
Spider Monkeys ◽  
Histopathological Analysis ◽  
Periodontal Pathogens ◽  
Periodontal Tissues ◽  
In Captivity

Background: Control of oral lesions contributes directly to the survival, and welfare of captive animals, and studies show that the genus Ateles has a higher prevalence of widespread periodontal disease compared to other genera. Anaerobic microbial species, considered as periodontal pathogens, are part of the biofilm community that contributes to the development of periodontitis. The present study aimed to detect periodontopathogenos in the oral cavity of two captive white-cheeked spider monkeys (Ateles marginatus) submitted for assessment oral and subgingival curettage.Case: We evaluated one pair of captive white-cheeked spider monkeys, one male (A) and one female (B), of 15 years of age with an average weight of 7 kg. Animals were fed daily with rations for primates, including fruit, vegetables, and raw eggs. The animals underwent oral evaluation, and following the charting of odontogram and photographic documentation, both were classified with periodontal disease stage III, according to the AVDC (American College of Veterinary Dentistry). They presented with moderate periodontitis, characterized by a loss of 25 to 50% of periodontal insertion and exposure of furcation degree 2, measured through clinical survey. During intraoral review, animals underwent subgingival curettage with curette of Gracey on the surface of the canine vestibular (C) and four top bilateral premolars (4PM). Antibiotics were not used at the time of collection, for dealing with routine procedures of clinical evaluation. The animals showed an increase in the volume of hemorrhagic features in the vestibular region between C and the second pre molar (2PM) on the upper right. Incisional biopsy was collected immediately at the end of the assessment, for the purpose of histopathological analyses. The samples from subgingival collection were immediately deposited in microtubes containing 500 µL of 0.9% saline solution and kept at -18°C until the time of genomic DNA extraction. The extracted DNA were subjected to polymerase chain reaction (PCR) for the detection of Fusobacterium spp., Porphyromonas gingivalis, Prevotella intermedia and Tannerella forsythia. All the samples detected in PCR were confirmed by partial sequencing of genes. Fusobacterium spp., P. gingivalis, and T. forsythia were detected in both individuals A and B. The identification of P. intermedia was exclusive to animal A and histopathological analysis of the gingival fragment showed severe and diffuse neutrophilic infiltrates, confirming the of gingivitis for this individual, with greater severity in the disease progression.Discussion: Older animals in captivity have a higher incidence of periodontal disease and high expression of inflammatory cytokines and immune receptors involved in the induction of inflammation as compared to younger animals, as well as free-living animals. Gingivitis and periodontitis are associated with an increase in the frequency of anaerobic, gramnegative microorganisms, whereby the production of pigments and a variety of enzymes damages the periodontal tissues. Knowing that the presence of bacterial species directly affects the periodontal condition and that the participation of these microorganisms favors the installation and progression of the disease, this study describes the first molecular detection of periodontopathogens related to periodontal disease in white-cheeked spider monkeys. We concluded that these pathogens are present in the oral cavity of white-cheeked spider monkeys, highlighting the importance of routine and proper food handling in captive animals.

scholarly journals Influence of Eugenia uniflora Extract on Adhesion to Human Buccal Epithelial Cells, Biofilm Formation, and Cell Surface Hydrophobicity of Candida spp. from the Oral Cavity of Kidney Transplant Recipients

Molecules ◽  
2018 ◽  
Vol 23 (10) ◽  
pp. 2418 ◽  
Author(s):  
Luanda Souza ◽  
Walicyranison Silva-Rocha ◽  
Magda Ferreira ◽  
Luiz Soares ◽  
Terezinha Svidzinski ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Epithelial Cells ◽  
Cell Surface ◽  
Oral Cavity ◽  
Kidney Transplant ◽  
Biofilm Formation ◽  
Cell Surface Hydrophobicity ◽  
Surface Hydrophobicity ◽  
Candida Spp ◽  
Buccal Epithelial Cells

This study evaluated the influence of the extract of Eugenia uniflora in adhesion to human buccal epithelial cells (HBEC) biofilm formation and cell surface hydrophobicity (CSH) of Candida spp. isolated from the oral cavity of kidney transplant patients. To evaluate virulence attributes in vitro, nine yeasts were grown in the presence and absence of 1000 μg/mL of the extract. Adhesion was quantified using the number of Candida cells adhered to 150 HBEC determined by optical microscope. Biofilm formation was evaluated using two methodologies: XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) and crystal violet assay, and further analyzed by electronic scan microscopy. CSH was quantified with the microbial adhesion to hydrocarbons test. We could detect that the extract of E. uniflora was able to reduce adhesion to HBEC and CSH for both Candida albicans and non-Candida albicans Candida species. We also observed a statistically significant reduced ability to form biofilms in biofilm-producing strains using both methods of quantification. However, two highly biofilm-producing strains of Candida tropicalis had a very large reduction in biofilm formation. This study reinforces the idea that besides growth inhibition, E. uniflora may interfere with the expression of some virulence factors of Candida spp. and may be possibly applied in the future as a novel antifungal agent.

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