scholarly journals High Biofilm Formation of Non-Smooth Candida parapsilosis Correlates with Increased Incorporation of GPI-Modified Wall Adhesins

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 493
Author(s):  
Ana Esther Moreno-Martínez ◽  
Emilia Gómez-Molero ◽  
Pablo Sánchez-Virosta ◽  
Henk L. Dekker ◽  
Albert de Boer ◽  
...  
Keyword(s):  
Cell Wall ◽  
Biofilm Formation ◽  
Candida Parapsilosis ◽  
Surface Expression ◽  
Growth Conditions ◽  
Clinical Isolates ◽  
Abiotic Surfaces ◽  
Abundance And Diversity ◽  
Cellular Phenotypes ◽  
Comprehensive Study

Candida parapsilosis is among the most frequent causes of candidiasis. Clinical isolates of this species show large variations in colony morphotype, ranging from round and smooth to a variety of non-smooth irregular colony shapes. A non-smooth appearance is related to increased formation of pseudohyphae, higher capacity to form biofilms on abiotic surfaces, and invading agar. Here, we present a comprehensive study of the cell wall proteome of C. parapsilosis reference strain CDC317 and seven clinical isolates under planktonic and sessile conditions. This analysis resulted in the identification of 40 wall proteins, most of them homologs of known Candida albicans cell wall proteins, such as Gas, Crh, Bgl2, Cht2, Ecm33, Sap, Sod, Plb, Pir, Pga30, Pga59, and adhesin family members. Comparative analysis of exponentially growing and stationary phase planktonic cultures of CDC317 at 30 °C and 37 °C revealed only minor variations. However, comparison of smooth isolates to non-smooth isolates with high biofilm formation capacity showed an increase in abundance and diversity of putative wall adhesins from Als, Iff/Hyr, and Hwp families in the latter. This difference depended more strongly on strain phenotype than on the growth conditions, as it was observed in planktonic as well as biofilm cells. Thus, in the set of isolates analyzed, the high biofilm formation capacity of non-smooth C. parapsilosis isolates with elongated cellular phenotypes correlates with the increased surface expression of putative wall adhesins in accordance with their proposed cellular function.

scholarly journals Characterization of Biofilm Formation and the Role of BCR1 in Clinical Isolates of Candida parapsilosis

2013 ◽  
Vol 13 (4) ◽  
pp. 438-451 ◽  
Author(s):  
Srisuda Pannanusorn ◽  
Bernardo Ramírez-Zavala ◽  
Heinrich Lünsdorf ◽  
Birgitta Agerberth ◽  
Joachim Morschhäuser ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Candida Parapsilosis ◽  
Clinical Isolates ◽  
Wild Type ◽  
Content Type ◽  
Initial Adhesion ◽  
High Level ◽  
Increased Susceptibility

ABSTRACT In Candida parapsilosis , biofilm formation is considered to be a major virulence factor. Previously, we determined the ability of 33 clinical isolates causing bloodstream infection to form biofilms and identified three distinct groups of biofilm-forming strains (negative, low, and high). Here, we establish two different biofilm structures among strains forming large amounts of biofilm in which strains with complex spider-like structures formed robust biofilms on different surface materials with increased resistance to fluconazole. Surprisingly, the transcription factor Bcr1, required for biofilm formation in Candida albicans and C. parapsilosis , has an essential role only in strains with low capacity for biofilm formation. Although BCR1 leads to the formation of more and longer pseudohyphae, it was not required for initial adhesion and formation of mature biofilms in strains with a high level of biofilm formation. Furthermore, an additional phenotype affected by BCR1 was the switch in colony morphology from rough to crepe, but only in strains forming high levels of biofilm. All bcr1 Δ/Δ mutants showed increased proteolytic activity and increased susceptibility to the antimicrobial peptides protamine and RP-1 compared to corresponding wild-type and complemented strains. Taken together, our results demonstrate that biofilm formation in clinical isolates of C. parapsilosis is both dependent and independent of BCR1 , but even in strains which showed a BCR1 -independent biofilm phenotype, BCR1 has alternative physiological functions.

scholarly journals Correlation between Biofilm Formation and the Hypoxic Response in Candida parapsilosis

2009 ◽  
Vol 8 (4) ◽  
pp. 550-559 ◽  
Author(s):  
Tristan Rossignol ◽  
Chen Ding ◽  
Alessandro Guida ◽  
Christophe d'Enfert ◽  
Desmond G. Higgins ◽  
...  
Keyword(s):  
Cell Wall ◽  
Biofilm Formation ◽  
Candida Parapsilosis ◽  
Transcriptional Profile ◽  
Open Reading Frames ◽  
Hypoxic Response ◽  
Hypoxic Conditions ◽  
Expression Of Genes ◽  
Biofilm Development ◽  
Reading Frames

ABSTRACT The ability of Candida parapsilosis to form biofilms on indwelling medical devices is correlated with virulence. To identify genes that are important for biofilm formation, we used arrays representing approximately 4,000 open reading frames (ORFs) to compare the transcriptional profile of biofilm cells growing in a microfermentor under continuous flow conditions with that of cells in planktonic culture. The expression of genes involved in fatty acid and ergosterol metabolism and in glycolysis, is upregulated in biofilms. The transcriptional profile of C. parapsilosis biofilm cells resembles that of Candida albicans cells grown under hypoxic conditions. We therefore subsequently used whole-genome arrays (representing 5,900 ORFs) to determine the hypoxic response of C. parapsilosis and showed that the levels of expression of genes involved in the ergosterol and glycolytic pathways, together with several cell wall genes, are increased. Our results indicate that there is substantial overlap between the hypoxic responses of C. parapsilosis and C. albicans and that this may be important for biofilm development. Knocking out an ortholog of the cell wall gene RBT1, whose expression is induced both in biofilms and under conditions of hypoxia in C. parapsilosis, reduces biofilm development.

scholarly journals Cellular Localization of Carbonic Anhydrase Nce103p in Candida albicans and Candida parapsilosis

2020 ◽  
Vol 21 (3) ◽  
pp. 850
Author(s):  
Jiří Dostál ◽  
Jan Blaha ◽  
Romana Hadravová ◽  
Martin Hubálek ◽  
Olga Heidingsfeld ◽  
...  
Keyword(s):  
Cell Wall ◽  
Candida Albicans ◽  
Carbonic Anhydrase ◽  
Cellular Localization ◽  
Candida Parapsilosis ◽  
Growth Conditions ◽  
Carbonic Anhydrases ◽  
Atmospheric Conditions ◽  
Low Co2 ◽  
Mitochondrial Fractions

Pathogenic yeasts Candida albicans and Candida parapsilosis possess a ß-type carbonic anhydrase Nce103p, which is involved in CO2 hydration and signaling. C. albicans lacking Nce103p cannot survive in low CO2 concentrations, e.g., in atmospheric growth conditions. Candida carbonic anhydrases are orthologous to the Saccharomyces cerevisiae enzyme, which had originally been detected as a substrate of a non-classical export pathway. However, experimental evidence on localization of C. albicans and C. parapsilosis carbonic anhydrases has not been reported to date. Immunogold labeling and electron microscopy used in the present study showed that carbonic anhydrases are localized in the cell wall and plasmatic membrane of both Candida species. This localization was confirmed by Western blot and mass spectrometry analyses of isolated cell wall and plasma membrane fractions. Further analysis of C. albicans and C. parapsilosis subcellular fractions revealed presence of carbonic anhydrases also in the cytosolic and mitochondrial fractions of Candida cells cultivated in shaken liquid cultures, under the atmospheric conditions.

scholarly journals Impact of manganese on biofilm formation and cell morphology of Candida parapsilosis clinical isolates with different biofilm forming abilities

2019 ◽  
Vol 19 (6) ◽  
Author(s):  
Sulman Shafeeq ◽  
Srisuda Pannanusorn ◽  
Youssef Elsharabasy ◽  
Bernardo Ramírez-Zavala ◽  
Joachim Morschhäuser ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Cell Morphology ◽  
Biofilm Formation ◽  
Candida Parapsilosis ◽  
Divalent Cations ◽  
Clinical Isolates ◽  
Human Pathogen ◽  
Commensal Species ◽  
Developmental Role ◽  
The Impact

ABSTRACT The commensal species Candida parapsilosis is an emerging human pathogen that has the ability to form biofilms. In this study, we explored the impact of the divalent cations cobalt (Co2+), copper (Cu2+), iron (Fe3+), manganese (Mn2+), nickel (Ni2+) and zinc (Zn2+) on biofilm formation of clinical isolates of C. parapsilosis with no, low and high biofilm forming abilities at 30 and 37°C. All strains besides one isolate showed a concentration-dependent enhancement of biofilm formation at 30°C in the presence of Mn2+ with a maximum at 2 mM. The biofilm forming ability of no and low biofilm forming isolates was >2-fold enhanced in the presence of 2 mM Mn2+, while the effect in high biofilm forming isolate was significantly less pronounced. Of note, cells in the biofilms of no and low biofilm forming strains differentiated into yeast and pseudohyphal cells similar in morphology to high biofilm formers. The biofilm transcriptional activator BCR1 has a dual developmental role in the absence and presence of 2 mM Mn2+ as it promoted biofilm formation of no biofilm forming strains, and, surprisingly, suppressed cells of no biofilm forming strains to develop into pseudohyphae and/or hyphae. Thus, environmental conditions can significantly affect the amount of biofilm formation and cell morphology of C. parapsilosis with Mn2+ to overcome developmental blocks to trigger biofilm formation and to partially relieve BCR1 suppressed cell differentiation.

scholarly journals Mat fimbriae promote biofilm formation by meningitis-associated Escherichia coli

Microbiology ◽  
2010 ◽  
Vol 156 (8) ◽  
pp. 2408-2417 ◽  
Author(s):  
Timo A. Lehti ◽  
Philippe Bauchart ◽  
Johanna Heikkinen ◽  
Jörg Hacker ◽  
Timo K. Korhonen ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Early Stage ◽  
Surface Expression ◽  
Growth Media ◽  
Biofilm Growth ◽  
Abiotic Surfaces ◽  
K 12 ◽  
Biofilm Development

The mat (or ecp) fimbrial operon is ubiquitous and conserved in Escherichia coli, but its functions remain poorly described. In routine growth media newborn meningitis isolates of E. coli express the meningitis-associated and temperature-regulated (Mat) fimbria, also termed E. coli common pilus (ECP), at 20 °C, and here we show that the six-gene (matABCDEF)-encoded Mat fimbria is needed for temperature-dependent biofilm formation on abiotic surfaces. The matBCDEF deletion mutant of meningitis E. coli IHE 3034 was defective in an early stage of biofilm development and consequently unable to establish a detectable biofilm, contrasting with IHE 3034 derivatives deleted for flagella, type 1 fimbriae or S-fimbriae, which retained the wild-type biofilm phenotype. Furthermore, induced production of Mat fimbriae from expression plasmids enabled biofilm-deficient E. coli K-12 cells to form biofilm at 20 °C. No biofilm was detected with IHE 3034 or MG1655 strains grown at 37 °C. The surface expression of Mat fimbriae and the frequency of Mat-positive cells in the IHE 3034 population from 20 °C were high and remained unaltered during the transition from planktonic to biofilm growth and within the matured biofilm community. Considering the prevalence of the highly conserved mat locus in E. coli genomes, we hypothesize that Mat fimbria-mediated biofilm formation is an ancestral characteristic of E. coli.

scholarly journals Biofilm Formation in a Hydrodynamic Environment by Novel FimH Variants and Ramifications for Virulence

2001 ◽  
Vol 69 (3) ◽  
pp. 1322-1328 ◽  
Author(s):  
Mark A. Schembri ◽  
Per Klemm
Keyword(s):  
Urinary Tract ◽  
Biofilm Formation ◽  
Random Mutagenesis ◽  
Cumulative Effect ◽  
Growth Conditions ◽  
E Coli ◽  
Enrichment Technique ◽  
Naturally Occurring ◽  
Abiotic Surfaces ◽  
Hydrodynamic Environment

ABSTRACT Type 1 fimbriae are surface-located adhesion organelles ofEscherichia coli that are directly associated with virulence of the urinary tract. They mediated-mannose-sensitive binding to different host surfaces by way of the minor fimbrial component FimH. Naturally occurring variants of FimH that bind strongly to terminally exposed monomannose residues have been associated with a pathogenicity-adaptive phenotype that enhances E. coli colonization of extraintestinal locations such as the urinary tract. The FimH adhesin also promotes biofilm formation in a mannose-inhibitable manner on abiotic surfaces under static growth conditions. In this study, we used random mutagenesis combined with a novel selection-enrichment technique to specifically identify mutations in the FimH adhesin that confer onE. coli the ability to form biofilms under hydrodynamic flow (HDF) conditions. We identified three FimH variants from our mutant library that could mediate an HDF biofilm formation phenotype to various degrees. This phenotype was induced by the cumulative effect of multiple changes throughout the receptor-binding region of the protein. Two of the HDF biofilm-forming FimH variants were insensitive to mannose inhibition and represent novel phenotypes not previously identified in naturally occurring isolates. Characterization of our enriched clones revealed some similarities to amino acid alterations that occur in urinary tract infection (UTI) strains. Subsequent screening of a selection of UTI FimH variants demonstrated that they too could promote biofilm formation on abiotic surfaces under HDF conditions. Interestingly, the same correlation was not observed for commensal FimH variants. FimH is a multifaceted protein prone to rapid microevolution. In addition to its previously documented roles in adherence and invasion, we have now demonstrated its function in biofilm formation on abiotic surfaces subjected to HDF conditions. The study indicates that UTI FimH variants possess adaptations that enhance biofilm formation and suggests a novel role for FimH in UTIs associated with medical implants such as catheters.

scholarly journals Temperature, pH and Trimethoprim-Sulfamethoxazole Are Potent Inhibitors of Biofilm Formation by Stenotrophomonas maltophilia Clinical Isolates

2017 ◽  
Vol 66 (4) ◽  
pp. 433-438 ◽  
Author(s):  
Marjan Biočanin ◽  
Haowa Madi ◽  
Zorica Vasiljević ◽  
Milan Kojić ◽  
Branko Jovčić ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Stenotrophomonas Maltophilia ◽  
Tertiary Care ◽  
Opportunistic Pathogen ◽  
Growth Conditions ◽  
Clinical Isolates ◽  
Multiple Drug ◽  
Dependent Manner ◽  
Virulence Determinants ◽  
Intrinsic Resistance

Stenotrophomonas maltophilia, an opportunistic pathogen usually connected with healthcare-associated infections, is an environmental bacterium. Intrinsic resistance to multiple antibiotics, with different virulence determinants in the last decade classified this bacterium in the group of global multiple drug resistant (MDR) organism. S. maltophilia clinical isolates, were collected from tertiary care pediatric hospital in Belgrade, Serbia to investigate influence of different factors on biofilm formation, kinetics of biofilm formation for strong biofilm producers and effect of trimethoprim-sulfamethoxazole (TMP/SMX) on formed biofilm. Most of the isolates (89.8%) were able to form a biofilm. Analysis of biofilm formation in different growth conditions showed that changing of temeperature and pH had the stronggest effect on biofilm formation almost equally in group of cystic fibrosis (CF) and non-CF strains. TMP/SMX in concentration of 50 μg/ml reduced completely 24 h old biofilms while concentration of 25 μg/ml effects formed biofilms in a strain dependent manner. Among strains able to form strong biofilm CF isolates formed biofilm slower than non-CF isolates, while shaking conditions did not affect biofilm formation. Swimming motility was detected in both CF and non-CF isolates, however more motile strain formed stronger biofilms. This study suggests that temperature, pH and TMP/SMX had the strongest influence on biofilm formation in analyzed collection of S. maltophilia. A positive correlation between motility and strength of formed biofilm was demonstrated.

scholarly journals O-Mannosylation in Candida albicans Enables Development of Interkingdom Biofilm Communities

mBio ◽  
2014 ◽  
Vol 5 (2) ◽  
Author(s):  
Lindsay C. Dutton ◽  
Angela H. Nobbs ◽  
Katy Jepson ◽  
Mark A. Jepson ◽  
M. Margaret Vickerman ◽  
...  
Keyword(s):  
Cell Wall ◽  
Candida Albicans ◽  
Biofilm Formation ◽  
Early Stage ◽  
Growth Conditions ◽  
Wild Type ◽  
Content Type ◽  
Subsequent Performance ◽  
Biofilm Development ◽  
Mutant Cells

ABSTRACTCandida albicansis a fungus that colonizes oral cavity surfaces, the gut, and the genital tract.Streptococcus gordoniiis a ubiquitous oral bacterium that has been shown to form biofilm communities withC. albicans. Formation of dual-speciesS. gordonii-C. albicansbiofilm communities involves interaction of theS. gordoniiSspB protein with the Als3 protein on the hyphal filament surface ofC. albicans. Mannoproteins comprise a major component of theC. albicanscell wall, and in this study we sought to determine if mannosylation in cell wall biogenesis ofC. albicanswas necessary for hyphal adhesin functions associated with interkingdom biofilm development. AC. albicans mnt1Δmnt2Δ mutant, with deleted α-1,2-mannosyltransferase genes and thus defective inO-mannosylation, was abrogated in biofilm formation under various growth conditions and produced hyphal filaments that were not recognized byS. gordonii. Cell wall proteomes of hypha-formingmnt1Δmnt2Δ mutant cells showed growth medium-dependent alterations, compared to findings for the wild type, in a range of protein components, including Als1, Als3, Rbt1, Scw1, and Sap9. Hyphal filaments formed bymnt1Δmnt2Δ mutant cells, unlike wild-type hyphae, did not interact withC. albicansAls3 or Hwp1 partner cell wall proteins or withS. gordoniiSspB partner adhesin, suggesting defective functionality of adhesins on themnt1Δmnt2Δ mutant. These observations imply that early stageO-mannosylation is critical for activation of hyphal adhesin functions required for biofilm formation, recognition by bacteria such asS. gordonii, and microbial community development.IMPORTANCEIn the human mouth, microorganisms form communities known as biofilms that adhere to the surfaces present.Candida albicansis a fungus that is often found within these biofilms. We have focused on the mechanisms by whichC. albicansbecomes incorporated into communities containing bacteria, such asStreptococcus. We find that impairment of early stage addition of mannose sugars toC. albicanshyphal filament proteins deleteriously affects their subsequent performance in mediating formation of polymicrobial biofilms. Our analyses provide new understanding of the way that microbial communities develop, and of potential means to controlC. albicansinfections.

scholarly journals Microbicides Alter the Expression and Function of RND-Type Efflux Pump AdeABC in Biofilm-Associated Cells of Acinetobacter baumannii Clinical Isolates

2015 ◽  
Vol 60 (1) ◽  
pp. 57-63 ◽  
Author(s):  
Suvarna Krishnamoorthy ◽  
Bhavikkumar P. Shah ◽  
Hiu Ham Lee ◽  
Luis R. Martinez
Keyword(s):  
Acinetobacter Baumannii ◽  
Biofilm Formation ◽  
Efflux Pump ◽  
Acquired Resistance ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Content Type ◽  
Abiotic Surfaces ◽  
Hospital Environments ◽  
And Function

ABSTRACTAcinetobacter baumanniiis a Gram-negative bacterium that causes nosocomial infections worldwide. This microbe's propensity to form biofilms allows it to persist and to survive on clinical abiotic surfaces for long periods. In fact,A. baumanniibiofilm formation and its multidrug-resistant nature severely compromise our capacity to care for patients in hospital environments. In contrast, microbicides such as cetrimide (CT) and chlorhexidine (CHX) play important roles in the prevention and treatment of infections. We assessed the efficacy of CT and CHX, either alone or in combination, in eradicatingA. baumanniibiofilms formed by clinical isolates, by using stainless steel washers to mimic hard abiotic surfaces found in hospital settings. We demonstrated that increasing amounts of each microbicide, alone or in combination, were able to damage and to reduce the viability ofA. baumanniibiofilms efficaciously. Interestingly, theadeBgene of the resistance-nodulation-cell division (RND) family is predominantly associated with acquired resistance to antimicrobials inA. baumannii. We showed that CT and CHX adversely modified the expression and function of the RND-type efflux pump AdeABC in biofilm-associatedA. baumanniicells. Furthermore, we established that these microbicides decreased the negative charges onA. baumanniicell membranes, causing dysregulation of the efflux pump and leading to cell death. Our findings suggest that CT and CHX, alone or in combination, can be used efficaciously for eradication ofA. baumanniifrom hospital surfaces, in order to reduce infections caused by this nosocomial agent.

scholarly journals Characterization of Campylobacter jejuni Biofilms under Defined Growth Conditions

2007 ◽  
Vol 73 (6) ◽  
pp. 1908-1913 ◽  
Author(s):  
Ryan J. Reeser ◽  
Robert T. Medler ◽  
Stephen J. Billington ◽  
B. Helen Jost ◽  
Lynn A. Joens
Keyword(s):  
Campylobacter Jejuni ◽  
Biofilm Formation ◽  
Broiler Chickens ◽  
Diarrheal Disease ◽  
Acrylonitrile Butadiene Styrene ◽  
Growth Conditions ◽  
Industrialized Countries ◽  
Abiotic Surfaces ◽  
Rich Media ◽  
Commercial Broiler

ABSTRACT Campylobacter jejuni is a major cause of human diarrheal disease in many industrialized countries and is a source of public health and economic burden. C. jejuni, present as normal flora in the intestinal tract of commercial broiler chickens and other livestock, is probably the main source of human infections. The presence of C. jejuni in biofilms found in animal production watering systems may play a role in the colonization of these animals. We have determined that C. jejuni can form biofilms on a variety of abiotic surfaces commonly used in watering systems, such as acrylonitrile butadiene styrene and polyvinyl chloride plastics. Furthermore, C. jejuni biofilm formation was inhibited by growth in nutrient-rich media or high osmolarity, and thermophilic and microaerophilic conditions enhanced biofilm formation. Thus, nutritional and environmental conditions affect the formation of C. jejuni biofilms. Both flagella and quorum sensing appear to be required for maximal biofilm formation, as C. jejuni flaAB and luxS mutants were significantly reduced in their ability to form biofilms compared to the wild-type strain.

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