Evaluating Metabolic Pathways and Biofilm Formation in Stenotrophomonas maltophilia

2021 ◽  
Author(s):  
Cierra M. Isom ◽  
Blake Fort ◽  
Gregory G. Anderson
Keyword(s):  
Amino Acids ◽  
Metabolic Pathways ◽  
Biofilm Formation ◽  
Stenotrophomonas Maltophilia ◽  
Glycolytic Enzyme ◽  
Phosphoglycerate Mutase ◽  
Biofilm Growth ◽  
Metabolic Activities ◽  
Biofilm Development ◽  
The Impact

Stenotrophomonas maltophilia has recently arisen as a prominent nosocomial pathogen because of its high antimicrobial resistance and ability to cause chronic respiratory infections. Often the infections are worsened by biofilm formation which enhances antibiotic tolerance. We have previously found that mutation of the gpmA gene, encoding the glycolytic enzyme phosphoglycerate mutase, impacts the formation of this biofilm on biotic and abiotic surfaces at early timepoints. This finding, indicating an association between carbon source and biofilm formation, led us to hypothesize that metabolism would influence S. maltophilia biofilm formation and planktonic growth. In the present study, we tested the impact of various growth substrates on biofilm levels and growth kinetics to determine metabolic requirements for these processes. We found that S. maltophilia wildtype preferred amino acids versus glucose for planktonic and biofilm growth and that gpmA deletion inhibited growth in amino acids. Furthermore, supplementation of the Δ gpmA strain by glucose or ribose phenotypically complemented growth defects. These results suggest that S. maltophilia shuttles amino acid carbon through gluconeogenesis to an undefined metabolic pathway supporting planktonic and biofilm growth. Further evaluation of these metabolic pathways might reveal novel metabolic activities of this pathogen. Importance Stenotrophomonas maltophilia is a prominent opportunistic pathogen that often forms biofilms during infection. However, the molecular mechanisms of virulence and biofilm formation are poorly understood. The glycolytic enzyme phosphoglycerate mutase appears to play a role in biofilm formation, and we used a mutant in its gene ( gpmA ) to probe the metabolic circuitry potentially involved in biofilm development. The results of our study indicate that S. maltophilia displays unique metabolic activities, which could be exploited for inhibiting growth and biofilm formation of this pathogen.

scholarly journals Effect of sodium silicate on drinking water biofilm development

2021 ◽  
Author(s):  
Sebastian Munoz ◽  
Benjamin F. Trueman ◽  
Bofu Li ◽  
Graham A. Gagnon
Keyword(s):  
Drinking Water ◽  
Sodium Silicate ◽  
Biofilm Formation ◽  
Corrosion Inhibitors ◽  
Biomass Accumulation ◽  
Corrosion Control ◽  
Biofilm Growth ◽  
Biofilm Development ◽  
The Impact ◽  
Sodium Silicates

AbstractSodium silicates have been studied for sequestration of iron, coagulation, and corrosion control, but their impact on biofilm formation has not been documented in detail. This study investigated the impact of sodium silicate corrosion control on biomass accumulation in drinking water systems in comparison to orthophosphate, a common corrosion inhibitor. Biofilm growth was measured by determining ATP concentrations, and the bacterial community was characterized using 16S ribosomal RNA (rRNA) sequencing. A pilot-scale study with cast-iron pipe loops, annular reactors (ARs), and polycarbonate coupons demonstrated significantly lower biofilm ATP concentrations in the sodium silicate-treated AR than the orthophosphate-treated AR when the water temperature exceeded 20°C. However, an elevated sodium silicate dose (48 mg L-1 of SiO2) disturbed and dispersed the biofilm formed inside the AR, resulting in elevated effluent ATP concentrations. Two separate experiments confirmed that biomass accumulation was higher in the presence of orthophosphate at high water temperatures (20°C) only. No significant differences were identified in biofilm ATP concentrations at lower water temperatures (below 20°C). Differences in bacterial communities between the orthophosphate- and sodium silicate-treated systems were not statistically significant, even though orthophosphate promoted higher biofilm growth. However, the genera Halomonas and Mycobacterium—which include opportunistic pathogens—were present at greater relative abundances in the orthophosphate-treated system compared to the sodium silicate system.Graphical abstractOrthophosphate promotes more biofilm growth in comparison to sodium silicates at water temperatures above 20°C.Water impact statementSodium silicates have been used in drinking water treatment for decades, both as sequestrants and as corrosion inhibitors. However, their impact on biofilm formation is poorly understood, and this risks drinking water quality. This study aims to further clarify the effects of corrosion inhibitors on biofilm development, including inhibitors that are not phosphate-based.

scholarly journals Improvement of XTT assay performance for studies involving Candida albicans biofilms

2008 ◽  
Vol 19 (4) ◽  
pp. 364-369 ◽  
Author(s):  
Wander José da Silva ◽  
Jayampath Seneviratne ◽  
Nipuna Parahitiyawa ◽  
Edvaldo Antonio Ribeiro Rosa ◽  
Lakshman Perera Samaranayake ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Biofilm Formation ◽  
Cell Activity ◽  
Growth Yield ◽  
Xtt Assay ◽  
Biofilm Growth ◽  
Assay Performance ◽  
Reduction Assay ◽  
Cell Layers ◽  
Biofilm Development

2, 3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) reduction assay has been used to study Candida biofilm formation. However, considering that the XTT reduction assay is dependent on cell activity, its use for evaluating mature biofilms may lead to inaccuracies since biofilm bottom cell layers tend to be relatively quiescent at later stages of biofilm formation. The aim of this study was to improve XTT reduction assay by adding glucose supplements to the standard XTT formulation. Candida albicans ATCC 90028 was used to form 24-, 48- and 72-h biofilms. The oxidative activity at 90, 180 and 270 min of incubation was evaluated. The control consisted of standard XTT formulation without glucose supplements, and was modified by the addition of 50, 100 and 200 mM of glucose. The XTT assay with 200 mM glucose showed more accurate and consistent readings correlating with biofilm development at 24, 48 and 72 h. Biofilm growth yield after 180 min incubation, when evaluated with the 200 mM glucose supplemented XTT, produced the most consistent readings on repetitive testing. It may be concluded that glucose supplementation of XTT could minimize variation and produce more accurate data for the XTT assay.

scholarly journals Characterization of Temporal Protein Production in Pseudomonas aeruginosa Biofilms

2005 ◽  
Vol 187 (23) ◽  
pp. 8114-8126 ◽  
Author(s):  
Christopher J. Southey-Pillig ◽  
David G. Davies ◽  
Karin Sauer
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antibiotic Resistance ◽  
Biofilm Formation ◽  
Protein Production ◽  
Developmental Stages ◽  
Developmental Process ◽  
Developmental Cycle ◽  
Specific Proteins ◽  
Biofilm Development ◽  
The Impact

ABSTRACT Phenotypic and genetic evidence supporting the notion of biofilm formation as a developmental process is growing. In the present work, we provide additional support for this hypothesis by identifying the onset of accumulation of biofilm-stage specific proteins during Pseudomonas aeruginosa biofilm maturation and by tracking the abundance of these proteins in planktonic and three biofilm developmental stages. The onset of protein production was found to correlate with the progression of biofilms in developmental stages. Protein identification revealed that proteins with similar function grouped within similar protein abundance patterns. Metabolic and housekeeping proteins were found to group within a pattern separate from virulence, antibiotic resistance, and quorum-sensing-related proteins. The latter were produced in a progressive manner, indicating that attendant features that are characteristic of biofilms such as antibiotic resistance and virulence may be part of the biofilm developmental process. Mutations in genes for selected proteins from several protein production patterns were made, and the impact of these mutations on biofilm development was evaluated. The proteins cytochrome c oxidase, a probable chemotaxis transducer, a two-component response regulator, and MexH were produced only in mature and late-stage biofilms. Mutations in the genes encoding these proteins did not confer defects in growth, initial attachment, early biofilm formation, or twitching motility but were observed to arrest biofilm development at the stage of cell cluster formation we call the maturation-1 stage. The results indicated that expression of theses genes was required for the progression of biofilms into three-dimensional structures on abiotic surfaces and the completion of the biofilm developmental cycle. Reverse transcription-PCR analysis confirmed the detectable change in expression of the respective genes ccoO, PA4101, and PA4208. We propose a possible mechanism for the role of these biofilm-specific proteins in biofilm formation.

scholarly journals The Relative Importance of Shear Forces and Surface Hydrophobicity on Biofilm Formation by Coccoid Cyanobacteria

Polymers ◽  
2020 ◽  
Vol 12 (3) ◽  
pp. 653 ◽  
Author(s):  
Sara I. Faria ◽  
Rita Teixeira-Santos ◽  
Maria J. Romeu ◽  
João Morais ◽  
Vitor Vasconcelos ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Hydrophobic Surface ◽  
Surface Hydrophobicity ◽  
Polymeric Coating ◽  
Relative Importance ◽  
Hydrodynamic Conditions ◽  
Shear Forces ◽  
Lower Shear ◽  
Biofilm Development ◽  
The Impact

Understanding the conditions affecting cyanobacterial biofilm development is crucial to develop new antibiofouling strategies and decrease the economic and environmental impact of biofilms in marine settings. In this study, we investigated the relative importance of shear forces and surface hydrophobicity on biofilm development by two coccoid cyanobacteria with different biofilm formation capacities. The strong biofilm-forming Synechocystis salina was used along with the weaker biofilm-forming Cyanobium sp. Biofilms were developed in defined hydrodynamic conditions using glass (a model hydrophilic surface) and a polymeric epoxy coating (a hydrophobic surface) as substrates. Biofilms developed in both surfaces at lower shear conditions contained a higher number of cells and presented higher values for wet weight, thickness, and chlorophyll a content. The impact of hydrodynamics on biofilm development was generally stronger than the impact of surface hydrophobicity, but a combined effect of these two parameters strongly affected biofilm formation for the weaker biofilm-producing organism. The antibiofilm performance of the polymeric coating was confirmed at the hydrodynamic conditions prevailing in ports. Shear forces were shown to have a profound impact on biofilm development in marine settings regardless of the fouling capacity of the existing flora and the hydrophobicity of the surface.

Biofilm formation by a biotechnologically important tropical marine yeast isolate, Yarrowia lipolytica NCIM 3589

2008 ◽  
Vol 58 (6) ◽  
pp. 1221-1229 ◽  
Author(s):  
D. H. Dusane ◽  
Y. V. Nancharaiah ◽  
V. P. Venugopalan ◽  
A. R. Kumar ◽  
S. S. Zinjarde
Keyword(s):  
Yarrowia Lipolytica ◽  
Biofilm Formation ◽  
Edible Oils ◽  
Carbon Sources ◽  
Three Dimensional ◽  
Ecological Niches ◽  
Lag Phase ◽  
Biofilm Growth ◽  
Water Media ◽  
Biofilm Development

Biofilm formation by Yarrowia lipolytica, a biotechnologically important fungus in microtitre plates, on glass slide surfaces and in flow cell was investigated. In microtitre plates, there was a short lag phase of adhesion followed by a period of rapid biofilm growth. The fungus formed extensive biofilms on glass slides, whereas in flow-cells a multicellular, three-dimensional microcolony structure was observed. The isolate formed biofilms in seawater and in fresh water media at neutral pH when grown in microtitre plates. The carbon sources differentially affected formation of biofilms in microtitre plates. Lactic acid, erythritol, glycerol, glucose and edible oils supported the formation of biofilms, while alkanes resulted in sub-optimal biofilm development. A variation in the morphology of the fungus was observed with different carbon sources. The results point to the possible existence of highly structured biofilms in varied ecological niches from where the yeast is isolated.

scholarly journals Architectural transitions in Vibrio cholerae biofilms at single-cell resolution

2016 ◽  
Vol 113 (14) ◽  
pp. E2066-E2072 ◽  
Author(s):  
Knut Drescher ◽  
Jörn Dunkel ◽  
Carey D. Nadell ◽  
Sven van Teeffelen ◽  
Ivan Grnja ◽  
...  
Keyword(s):  
Single Cell ◽  
Vibrio Cholerae ◽  
Biofilm Formation ◽  
Bacterial Species ◽  
Bacterial Biomass ◽  
Biofilm Growth ◽  
Glass Substrates ◽  
Critical Transitions ◽  
Cell Shapes ◽  
Biofilm Development

Many bacterial species colonize surfaces and form dense 3D structures, known as biofilms, which are highly tolerant to antibiotics and constitute one of the major forms of bacterial biomass on Earth. Bacterial biofilms display remarkable changes during their development from initial attachment to maturity, yet the cellular architecture that gives rise to collective biofilm morphology during growth is largely unknown. Here, we use high-resolution optical microscopy to image all individual cells in Vibrio cholerae biofilms at different stages of development, including colonies that range in size from 2 to 4,500 cells. From these data, we extracted the precise 3D cellular arrangements, cell shapes, sizes, and global morphological features during biofilm growth on submerged glass substrates under flow. We discovered several critical transitions of the internal and external biofilm architectures that separate the major phases of V. cholerae biofilm growth. Optical imaging of biofilms with single-cell resolution provides a new window into biofilm formation that will prove invaluable to understanding the mechanics underlying biofilm development.

scholarly journals Phenotypic Characterization of Streptococcus pneumoniae Biofilm Development

2006 ◽  
Vol 188 (7) ◽  
pp. 2325-2335 ◽  
Author(s):  
Magee Allegrucci ◽  
F. Z. Hu ◽  
K. Shen ◽  
J. Hayes ◽  
Garth D. Ehrlich ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Biofilm Formation ◽  
Protein Identification ◽  
Development Process ◽  
Developmental Stages ◽  
Developmental Process ◽  
Phenotypic Characterization ◽  
Biofilm Growth ◽  
Cell Counts ◽  
Biofilm Development

ABSTRACT Streptococcus pneumoniae is among the most common pathogens associated with chronic otitis media with effusion, which has been hypothesized to be a biofilm disease. S. pneumoniae has been shown to form biofilms, however, little is known about the developmental process, the architecture, and the changes that occur upon biofilm development. In the current study we made use of a continuous-culture biofilm system to characterize biofilm development of 14 different S. pneumoniae strains representing at least 10 unique serotypes. The biofilm development process was found to occur in three distinct stages, including initial attachment, cluster formation, and biofilm maturation. While all 14 pneumococcal strains displayed similar developmental stages, the mature biofilm architecture differed significantly among the serotypes tested. Overall, three biofilm architectural groups were detected based on biomass, biofilm thickness, and cluster size. The biofilm viable cell counts and total protein concentration increased steadily over the course of biofilm development, reaching ∼8 × 108 cells and ∼15 mg of protein per biofilm after 9 days of biofilm growth. Proteomic analysis confirmed the presence of distinct biofilm developmental stages by the detection of multiple phenotypes over the course of biofilm development. The biofilm development process was found to correlate not only with differential production of proteins but also with a dramatic increase in the number of detectable proteins, indicating that biofilm formation by S. pneumoniae may be a far more complex process than previously anticipated. Protein identification revealed that proteins involved in virulence, adhesion, and resistance were more abundant under biofilm growth conditions. A possible role of the identified proteins in biofilm formation is discussed.

scholarly journals A Sustained-Release Membrane of Thiazolidinedione-8: Effect on Formation of a Candida/Bacteria Mixed Biofilm on Hydroxyapatite in a Continuous Flow Model

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Mark Feldman ◽  
Julia Shenderovich ◽  
Eran Lavy ◽  
Michael Friedman ◽  
Doron Steinberg
Keyword(s):  
Sustained Release ◽  
Biofilm Formation ◽  
Active Agent ◽  
Extracellular Polysaccharides ◽  
Flow Conditions ◽  
Biofilm Thickness ◽  
Release Drug ◽  
Flow Through ◽  
Biofilm Development ◽  
The Impact

Thiazolidinediones (TZDs) have been found to act as effective quorum sensing quenchers, capable of preventing biofilm formation. Our previous studies demonstrated a profound antibiofilm effect of the TZD derivative thiazolidinedione-8 (S-8), either in solution or incorporated into a sustained-release membrane (SRM-S-8) under batch conditions. In the present study, we used a constant depth film fermenter model in order to investigate the impact of SRM-S-8 on mixedC. albicans-S. mutansbiofilm development, under flow conditions. We found that essential parameters of cospecies biofilm maintenance and maturation, such as metabolic activity, biofilm thickness, roughness, extracellular polysaccharides production, and morphology of both pathogens, were altered by SRM-S-8 in the flow system. We propose that prolonged and sustained release of S-8 in a flow-through system allows better penetration of the active agent to deeper layers of the mixed biofilm, thereby increasing its activity against both pathogens. In conclusion, the use of a locally applied sustained-release drug delivery system of S-8 can affect the dental polymicrobial biofilm, resulting in clinical improvements and a better patient compliance.

Biofilm Formation by Candida Species on Silicone Surfaces and Latex Pacifier Nipples: An in vitro Study

2009 ◽  
Vol 33 (3) ◽  
pp. 235-240 ◽  
Author(s):  
Luiz Cezar da Silveira ◽  
Senda Charone ◽  
Lucianne Cople Maia ◽  
Rosangela Maria de Araújo Soares ◽  
Maristela Barbosa Portela
Keyword(s):  
Candida Species ◽  
Biofilm Formation ◽  
In Vitro Study ◽  
Reduction Reaction ◽  
Fungal Colonization ◽  
Candida Spp ◽  
Biofilm Growth ◽  
Species Analysis ◽  
Biofilm Development

The present study assessed the growth and development of biofilm formation by isolates of C. albicans, C. glabrata and C. parapsilosis on silicone and latex pacifier nipples. The silicone and latex surfaces were evaluated by scanning electronic microscopy (SEM). The plastic component of the nipple also seems to be an important factor regarding the biofilm formation by Candida spp. The biofilm growth was measured using the MTT reduction reaction. C. albicans was found to have a slightly greater capacity of forming biofilm compared to the other Candida species. Analysis of the pattern of biofilm development by C. albicans,C. glabrata and C. parapsilosis on latex and silicon pacifier shields showed an increased biofilm formation regarding the latter substrate. Silicone was shown to be more resistant to fungal colonization, particularly in the case of C. parapsilosis, despite the lack of any statistically significant differences (P > 0.05). In addition, silicone has a smoother surface compared to latex, whose surface was found to be rugose and irregular

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.

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