scholarly journals Comparison of the Genetic Features Involved in Bacillus subtilis Biofilm Formation Using Multi-Culturing Approaches

2021 ◽  
Vol 9 (3) ◽  
pp. 633
Author(s):  
Yasmine Dergham ◽  
Pilar Sanchez-Vizuete ◽  
Dominique Le Coq ◽  
Julien Deschamps ◽  
Arnaud Bridier ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Biofilm Formation ◽  
Negative Impact ◽  
Liquid Interface ◽  
Subtilis Strain ◽  
Pellicle Formation ◽  
Biofilm Model ◽  
Genetic Features ◽  
Semi Solid ◽  
Air Liquid Interface

Surface-associated multicellular assemblage is an important bacterial trait to withstand harsh environmental conditions. Bacillus subtilis is one of the most studied Gram-positive bacteria, serving as a model for the study of genetic pathways involved in the different steps of 3D biofilm formation. B. subtilis biofilm studies have mainly focused on pellicle formation at the air-liquid interface or complex macrocolonies formed on nutritive agar. However, only few studies focus on the genetic features of B. subtilis submerged biofilm formation and their link with other multicellular models at the air interface. NDmed, an undomesticated B. subtilis strain isolated from a hospital, has demonstrated the ability to produce highly structured immersed biofilms when compared to strains classically used for studying B. subtilis biofilms. In this contribution, we have conducted a multi-culturing comparison (between macrocolony, swarming, pellicle, and submerged biofilm) of B. subtilis multicellular communities using the NDmed strain and mutated derivatives for genes shown to be required for motility and biofilm formation in pellicle and macrocolony models. For the 15 mutated NDmed strains studied, all showed an altered phenotype for at least one of the different culture laboratory assays. Mutation of genes involved in matrix production (i.e., tasA, epsA-O, cap, ypqP) caused a negative impact on all biofilm phenotypes but favored swarming motility on semi-solid surfaces. Mutation of bslA, a gene coding for an amphiphilic protein, affected the stability of the pellicle at the air-liquid interface with no impact on the submerged biofilm model. Moreover, mutation of lytF, an autolysin gene required for cell separation, had a greater effect on the submerged biofilm model than that formed at aerial level, opposite to the observation for lytABC mutant. In addition, B. subtilis NDmed with sinR mutation formed wrinkled macrocolony, less than that formed by the wild type, but was unable to form neither thick pellicle nor structured submerged biofilm. The results are discussed in terms of the relevancy to determine whether genes involved in colony and pellicle formation also govern submerged biofilm formation, by regarding the specificities in each model.

Characterization of biofilm formation by Salmonella enterica at the air-liquid interface in aquatic environments

2018 ◽  
Vol 190 (4) ◽  
Author(s):  
José Andrés Medrano-Félix ◽  
Cristóbal Chaidez ◽  
Kristina D. Mena ◽  
María del Socorro Soto-Galindo ◽  
Nohelia Castro-del Campo
Keyword(s):  
Salmonella Enterica ◽  
Biofilm Formation ◽  
Liquid Interface ◽  
Aquatic Environments ◽  
Air Liquid Interface

scholarly journals The RapP-PhrP Quorum-Sensing System of Bacillus subtilis Strain NCIB3610 Affects Biofilm Formation through Multiple Targets, Due to an Atypical Signal-Insensitive Allele of RapP

2014 ◽  
Vol 197 (3) ◽  
pp. 592-602 ◽  
Author(s):  
Shira Omer Bendori ◽  
Shaul Pollak ◽  
Dorit Hizi ◽  
Avigdor Eldar
Keyword(s):  
Bacillus Subtilis ◽  
Quorum Sensing ◽  
Biofilm Formation ◽  
Response Regulator ◽  
Subtilis Strain ◽  
Transcriptional Level ◽  
Response Regulators ◽  
Multiple Targets ◽  
Content Type ◽  
Negative Effect

The genome ofBacillus subtilis168 encodes eightrap-phrquorum-sensing pairs. Rap proteins of all characterized Rap-Phr pairs inhibit the function of one or several important response regulators: ComA, Spo0F, or DegU. This inhibition is relieved upon binding of the peptide encoded by the cognatephrgene.Bacillus subtilisstrain NCIB3610, the biofilm-proficient ancestor of strain 168, encodes, in addition, therapP-phrPpair on the plasmid pBS32. RapP was shown to dephosphorylate Spo0F and to regulate biofilm formation, but unlike other Rap-Phr pairs, RapP does not interact with PhrP. In this work we extend the analysis of the RapP pathway by reexamining its transcriptional regulation, its effect on downstream targets, and its interaction with PhrP. At the transcriptional level, we show thatrapPandphrPregulation is similar to that of otherrap-phrpairs. We further find that RapP has an Spo0F-independent negative effect on biofilm-related genes, which is mediated by the response regulator ComA. Finally, we find that the insensitivity of RapP to PhrP is due to a substitution of a highly conserved residue in the peptide binding domain of therapPallele of strain NCIB3610. Reversing this substitution to the consensus amino acid restores the PhrP dependence of RapP activity and eliminates the effects of therapP-phrPlocus on ComA activity and biofilm formation. Taken together, our results suggest that RapP strongly represses biofilm formation through multiple targets and that PhrP does not counteract RapP due to a rare mutation inrapP.

scholarly journals Disruption of Escherichia coli Amyloid-Integrated Biofilm Formation at the Air–Liquid Interface by a Polysorbate Surfactant

Langmuir ◽  
2013 ◽  
Vol 29 (3) ◽  
pp. 920-926 ◽  
Author(s):  
Cynthia Wu ◽  
Ji Youn Lim ◽  
Gerald G. Fuller ◽  
Lynette Cegelski
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Liquid Interface ◽  
Air Liquid Interface

scholarly journals RemA (YlzA) and RemB (YaaB) Regulate Extracellular Matrix Operon Expression and Biofilm Formation in Bacillus subtilis

2009 ◽  
Vol 191 (12) ◽  
pp. 3981-3991 ◽  
Author(s):  
Jared T. Winkelman ◽  
Kris M. Blair ◽  
Daniel B. Kearns
Keyword(s):  
Extracellular Matrix ◽  
Bacillus Subtilis ◽  
Biofilm Formation ◽  
Extracellular Polysaccharide ◽  
Pellicle Formation ◽  
Multicellular Aggregates ◽  
The Matrix ◽  
Matrix Components ◽  
Operon Expression ◽  
Motility Inhibition

ABSTRACT Biofilms are multicellular aggregates stabilized by an extracellular matrix. In Bacillus subtilis, the biofilm matrix is composed of an extracellular polysaccharide and the secreted protein TasA. Expression of both of the matrix components is repressed by the DNA-binding master regulator, SinR. Here we identify two small protein regulators of the extracellular matrix: RemA (formerly YlzA) and RemB (formerly YaaB). Mutation of RemA or RemB impairs pellicle formation, complex colony architecture, and motility inhibition in a sinR mutant background. Both proteins are required for the activation of the matrix biosynthesis operons and appear to act in parallel to SinR and two other known biofilm regulators, AbrB and DegU.

scholarly journals Morphological and proteomic analysis of early stage air–liquid interface biofilm formation in Mycobacterium smegmatis

Microbiology ◽  
2014 ◽  
Vol 160 (7) ◽  
pp. 1346-1356 ◽  
Author(s):  
Zuzana Sochorová ◽  
Denisa Petráčková ◽  
Barbora Sitařová ◽  
Karolína Buriánková ◽  
Silvia Bezoušková ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Cell Wall ◽  
Mycobacterium Smegmatis ◽  
Expression Profiles ◽  
Liquid Interface ◽  
Pellicle Formation ◽  
Biofilm Development ◽  
Air Liquid Interface ◽  
Scanning Electron

We studied the early stages of pellicle formation by Mycobacterium smegmatis on the surface of a liquid medium [air–liquid interface (A–L)]. Using optical and scanning electron microscopy, we showed the formation of a compact biofilm pellicle from micro-colonies over a period of 8–30 h. The cells in the pellicle changed size and cell division pattern during this period. Based on our findings, we created a model of M. smegmatis A–L early pellicle formation showing the coordinate growth of cells in the micro-colonies and in the homogeneous film between them, where the accessibility to oxygen and nutrients is different. A proteomic approach utilizing high-resolution two-dimensional gel electrophoresis, in combination with mass spectrometry-based protein identification, was used to analyse the protein expression profiles of the different morphological stages of the pellicle. The proteins identified formed four expression groups; the most interesting of these groups contained the proteins with highest expression in the biofilm development phase, when the floating micro-colonies containing long and more robust cells associate into flocs and start to form a compact pellicle. The majority of these proteins, including GroEL1, are involved in cell wall synthesis or modification, mostly through the involvement of mycolic acid biosynthesis, and their expression maxima correlated with the changes in cell size and the rigidity of the bacterial cell wall observed by scanning electron microscopy.

scholarly journals Extending an Eco-Evolutionary Understanding of Biofilm-Formation at the Air-Liquid Interface to Community Biofilms

2020 ◽  
Author(s):  
Robyn Jerdan ◽  
Olga Iungin ◽  
Olena V. Moshynets ◽  
Geert Potters ◽  
Andrew J. Spiers
Keyword(s):  
Biofilm Formation ◽  
Liquid Interface ◽  
Air Liquid Interface

scholarly journals Biofilm Formation by and Multicellular Behavior of Escherichia coli O55:H7, an Atypical Enteropathogenic Strain

2010 ◽  
Vol 76 (5) ◽  
pp. 1545-1554 ◽  
Author(s):  
Michal Weiss-Muszkat ◽  
Dana Shakh ◽  
Yizhou Zhou ◽  
Riky Pinto ◽  
Eddy Belausov ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Genetic Relatedness ◽  
Liquid Interface ◽  
E Coli ◽  
Wild Type Phenotype ◽  
Enteropathogenic Strain ◽  
Biofilm Development ◽  
Air Liquid Interface ◽  
High Degree

ABSTRACT Enteropathogenic Escherichia coli (EPEC) is an important causal agent of diarrheal illness throughout the world. Nevertheless, researchers have only recently begun to explore its capacity to form biofilms. Strain O55:H7 (DMS9) is a clinical isolate belonging to the atypical EPEC (aEPEC) group, which displays a high degree of genetic relatedness to enterohemorrhagic E. coli. Strain DMS9 formed a robust biofilm on an abiotic surface at 26�C, but not at 37�C. It also formed a dense pellicle at the air-liquid interface and developed a red, rough, and dry (RDAR) morphotype on Congo red agar. Unlike a previously described E. coli O157:H7 strain, the aEPEC strain seems to express cellulose. Transposon mutagenesis was used to identify biofilm-deficient mutants. One of the mutants was inactivated in the csgFG genes, required for assembly and secretion of curli fimbriae, while a second mutant had a mutation in crl, a thermosensitive global regulator that modulates σS activity and downstream expression of curli and cellulose. The two mutants were deficient in their biofilm formation capabilities and did not form a pellicle at the air-liquid interface. Unlike in Salmonella, the csgFG mutant in aEPEC completely lost the RDAR phenotype, while the crl mutant displayed a unique RDAR “pizza”-like morphotype. Genetic complementation of the two mutants resulted in restoration of the wild-type phenotype. This report is the first to describe and analyze a multicellular behavior in aEPEC and support a major role for curli and the crl regulator in biofilm development at low temperatures corresponding to the nonmammalian host environment.

scholarly journals Biofilm formation by South African non-O157 Shiga toxigenic Escherichia coli on stainless steel coupons

2020 ◽  
Vol 66 (4) ◽  
pp. 328-336 ◽  
Author(s):  
Emmanuel W. Bumunang ◽  
Collins N. Ateba ◽  
Kim Stanford ◽  
Tim A. McAllister ◽  
Yan D. Niu
Keyword(s):  
Escherichia Coli ◽  
Stainless Steel ◽  
South African ◽  
Food Processing ◽  
Biofilm Formation ◽  
Viable Cell ◽  
Liquid Interface ◽  
Cell Counts ◽  
Potential Source ◽  
Air Liquid Interface

This study examined the biofilm-forming ability of six non-O157 Shiga-toxin-producing Escherichia coli (STEC) strains: O116:H21, wzx-Onovel5:H19, O129:H21, O129:H23, O26:H11, and O154:H10 on stainless steel coupons after 24, 48, and 72 h of incubation at 22 °C and after 168 h at 10 °C. The results of crystal violet staining revealed that strains O129:H23 and O154:H10 were able to form biofilms on both the submerged surface and the air–liquid interface of coupons, whereas strains O116:H21, wzx-Onovel5:H19, O129:H21, and O26:H11 formed biofilm only at the air–liquid interface. Viable cell counts and scanning electron microscopy showed that biofilm formation increased (p < 0.05) over time. The biofilm-forming ability of non-O157 STEC was strongest (p < 0.05) at 22 °C after 48 h of incubation. The strongest biofilm former regardless of temperature was O129:H23. Generally, at 10 °C, weak to no biofilm was observed for isolates O154:H10, O116:H21, wzx-Onovel5:H19, O26:H11, and O129:H21 after 168 h. This study found that temperature affected the biofilm-forming ability of non-O157 STEC strains. Overall, our data indicate a high potential for biofilm formation by the isolates at 22 °C, suggesting that non-O157 STEC strains could colonize stainless steel within food-processing facilities. This could serve as a potential source of adulteration and promote the dissemination of these potential pathogens in food.

scholarly journals Air-Liquid Interface Biofilms of Bacillus cereus: Formation, Sporulation, and Dispersion

2007 ◽  
Vol 73 (5) ◽  
pp. 1481-1488 ◽  
Author(s):  
Janneke G. E. Wijman ◽  
Patrick P. L. A. de Leeuw ◽  
Roy Moezelaar ◽  
Marcel H. Zwietering ◽  
Tjakko Abee
Keyword(s):  
Bacillus Cereus ◽  
Biofilm Formation ◽  
Culture Conditions ◽  
Liquid Interface ◽  
Production Cycle ◽  
Microtiter Plates ◽  
Production Environments ◽  
Piping Systems ◽  
Industrial Storage ◽  
Air Liquid Interface

ABSTRACT Biofilm formation by Bacillus cereus was assessed using 56 strains of B. cereus, including the two sequenced strains, ATCC 14579 and ATCC 10987. Biofilm production in microtiter plates was found to be strongly dependent on incubation time, temperature, and medium, as well as the strain used, with some strains showing biofilm formation within 24 h and subsequent dispersion within the next 24 h. A selection of strains was used for quantitative analysis of biofilm formation on stainless steel coupons. Thick biofilms of B. cereus developed at the air-liquid interface, while the amount of biofilm formed was much lower in submerged systems. This suggests that B. cereus biofilms may develop particularly in industrial storage and piping systems that are partly filled during operation or where residual liquid has remained after a production cycle. Moreover, depending on the strain and culture conditions, spores constituted up to 90% of the total biofilm counts. This indicates that B. cereus biofilms can act as a nidus for spore formation and subsequently can release their spores into food production environments.

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