CD25890, a conserved protein that modulates sporulation initiation in Clostridioides difficile

AbstractBacteria that reside in the gastrointestinal tract of healthy humans are essential for our health, sustenance and well-being. About 50–60% of those bacteria have the ability to produce resilient spores that are important for the life cycle in the gut and for host-to-host transmission. A genomic signature for sporulation in the human intestine was recently described, which spans both commensals and pathogens such as Clostridioides difficile and contains several genes of unknown function. We report on the characterization of a signature gene, CD25890, which, as we show is involved in the control of sporulation initiation in C. difficile under certain nutritional conditions. Spo0A is the main regulatory protein controlling entry into sporulation and we show that an in-frame deletion of CD25890 results in increased expression of spo0A per cell and increased sporulation. The effect of CD25890 on spo0A is likely indirect and mediated through repression of the sinRR´ operon. Deletion of the CD25890 gene, however, does not alter the expression of the genes coding for the cytotoxins or the genes involved in biofilm formation. Our results suggest that CD25890 acts to modulate sporulation in response to the nutrients present in the environment.

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A regulatory protein that represses sporulation in Clostridioides difficile

10.1101/2020.02.25.964569 ◽

2020 ◽

Author(s):

Diogo Martins ◽

Aristides L. Mendes ◽

Jessica Antunes ◽

Adriano O. Henriques ◽

Mónica Serrano

Keyword(s):

Regulatory Protein ◽

Well Being ◽

Toxin Production ◽

Sigma Factors ◽

Regulatory Proteins ◽

Proteolytic Activation ◽

Healthy Humans ◽

Clostridioides Difficile ◽

Sporulation Initiation

AbstractBacteria that reside in the gastrointestinal tract of healthy humans are essential for our health, sustenance and well-being. About 50 to 60% of those bacteria have the ability to produce resilient spores, important for the life cycle in the gut and for host-to-host transmission. A genomic signature for sporulation in the human intestine was recently described, which spans both commensals and pathogens such as Clostridioides difficile, and contains several genes of unknown function. We report on the characterization of a signature gene, csiA, which, as we show, is involved in the control of sporulation initiation in C. difficile. Spo0A is the main regulatory protein controlling entry into sporulation and we show that an in-frame deletion of csiA results in increased sporulation, and increased expression of spo0A per cell. Spo0A also drives transcription of the spoIIA and spoIIG operons, coding for the first forespore-(σF) and mother cell-specific (σE) RNA polymerase sigma factors. Strikingly, deletion of csiA increases expression of the spoIIG operon, but not that of the spoIIA operon. Increased expression of spoIIG results in increased production and proteolytic activation of pro-σE, suggesting that normally, the levels of active σE are limiting for sporulation. While other regulatory proteins affect both sporulation and several processes during the transition phase of growth, including toxin production or motility, deletion of the csiA gene does not alter the expression of the genes coding for the TcdA and TcdB cytotoxins or the genes involved in motility. Thus, our results establish that CsiA acts to modulate sporulation by reducing expression of the spo0A gene.

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c-di-GMP Inhibits Early Sporulation in Clostridioides difficile

mSphere ◽

10.1128/msphere.00919-21 ◽

2021 ◽

Author(s):

Adrianne N. Edwards ◽

Caitlin L. Willams ◽

Nivedita Pareek ◽

Shonna M. McBride ◽

Rita Tamayo

Keyword(s):

Life Cycle ◽

Biofilm Formation ◽

Toxin Production ◽

Cyclic Diguanylate ◽

Content Type ◽

Critical Step ◽

Clostridioides Difficile ◽

Physiological Processes ◽

Gastrointestinal Pathogen ◽

The Impact

Many bacterial organisms utilize the small signaling molecule cyclic diguanylate (c-di-GMP) to regulate important physiological processes, including motility, toxin production, biofilm formation, and colonization. c-di-GMP inhibits motility and toxin production and promotes biofilm formation and colonization in the anaerobic, gastrointestinal pathogen Clostridioides difficile . However, the impact of c-di-GMP on C. difficile spore formation, a critical step in this pathogen’s life cycle, is unknown.

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Immobilizing hydrolytic active Papain on biodegradable PLLA for biofilm inhibition in cardiovascular applications

Current Directions in Biomedical Engineering ◽

10.1515/cdbme-2020-3044 ◽

2020 ◽

Vol 6 (3) ◽

pp. 172-175

Author(s):

Michael Teske ◽

Tina Kießlich ◽

Niels Grabow ◽

Sabine Illner ◽

Julia Fischer ◽

...

Keyword(s):

Biofilm Formation ◽

Antimicrobial Agents ◽

Hydrolytic Enzyme ◽

Bacterial Biofilm ◽

Biofilm Inhibition ◽

Biofilm Growth ◽

Clostridioides Difficile ◽

Adhesive Surface

AbstractThe use of biomaterials in medicine is becoming increasingly important. One of the main concerns is the foreign body associated infection caused by direct microbial contamination or clinical infections. The bacterial biofilm formation on biomaterials depends on their surface properties. Therefore, several anti-adhesive surface modifications were developed. Nevertheless, the demand for antimicrobial agents that prevent bacterial colonisation is still largely unmet. The immobilization of active antimicrobial agents, such as antibacterial peptides or enzymes, offers a potential approach to achieve long-lasting effectiveness. In this investigation, the hydrolytic enzyme papain with its published antibacterial activity was covalently immobilized on the well-established biodegradable biomaterial poly-L-lactic acid (PLLA). For the characterization of the enzymes on the PLLA surfaces, the protein content and enzyme activity were determined. A biofilm assay was performed to test the effect of the papain-modified PLLA samples on the biofilm-forming bacterial strain Clostridioides difficile, one of the most frequently occurring human nosocomial pathogens. The investigated hydrolytic enzyme papain could be immobilized by coupling via the crosslinker EDC to the PLLA surface. Detection was performed by determination of the amount of protein and the reduced biofilm growth after 24 h and 72 h compared to the reference.

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Characterization of Altered Waking States of Consciousness in Healthy Humans

Case Medical Research ◽

10.31525/ct1-nct03853577 ◽

2019 ◽

Author(s):

Keyword(s):

Healthy Humans ◽

States Of Consciousness

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Faculty Opinions recommendation of Molecular characterization of host-specific biofilm formation in a vertebrate gut symbiont.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.718226618.793518484 ◽

2016 ◽

Author(s):

Heidi Goodrich-Blair

Keyword(s):

Molecular Characterization ◽

Biofilm Formation

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Phylogenomic analysis of Clostridioides difficile ribotype 106 strains reveals novel genetic islands and emergent phenotypes

Scientific Reports ◽

10.1038/s41598-020-79123-2 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Bryan Angelo P. Roxas ◽

Jennifer Lising Roxas ◽

Rachel Claus-Walker ◽

Anusha Harishankar ◽

Asad Mansoor ◽

...

Keyword(s):

Biofilm Formation ◽

Diarrheal Disease ◽

The United States ◽

Genomic Islands ◽

Rapid Expansion ◽

Phylogenomic Analysis ◽

Community Settings ◽

Hamster Model ◽

Clostridioides Difficile ◽

Healthcare Associated

AbstractClostridioides difficile infection (CDI) is a major healthcare-associated diarrheal disease. Consistent with trends across the United States, C. difficile RT106 was the second-most prevalent molecular type in our surveillance in Arizona from 2015 to 2018. A representative RT106 strain displayed robust virulence and 100% lethality in the hamster model of acute CDI. We identified a unique 46 KB genomic island (GI1) in all RT106 strains sequenced to date, including those in public databases. GI1 was not found in its entirety in any other C. difficile clade, or indeed, in any other microbial genome; however, smaller segments were detected in Enterococcus faecium strains. Molecular clock analyses suggested that GI1 was horizontally acquired and sequentially assembled over time. GI1 encodes homologs of VanZ and a SrtB-anchored collagen-binding adhesin, and correspondingly, all tested RT106 strains had increased teicoplanin resistance, and a majority displayed collagen-dependent biofilm formation. Two additional genomic islands (GI2 and GI3) were also present in a subset of RT106 strains. All three islands are predicted to encode mobile genetic elements as well as virulence factors. Emergent phenotypes associated with these genetic islands may have contributed to the relatively rapid expansion of RT106 in US healthcare and community settings.

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The Potential Role of Bacteriophages in the Treatment of Recalcitrant Chronic Rhinosinusitis

Antibiotics ◽

10.3390/antibiotics10060675 ◽

2021 ◽

Vol 10 (6) ◽

pp. 675

Author(s):

Saartje Uyttebroek ◽

Jolien Onsea ◽

Willem-Jan Metsemakers ◽

Lieven Dupont ◽

David Devolder ◽

...

Keyword(s):

Chronic Rhinosinusitis ◽

Biofilm Formation ◽

Potential Role ◽

Phage Therapy ◽

Well Being ◽

Treatment Alternative ◽

Surgical Interventions ◽

Social And Emotional ◽

Promising Treatment

Chronic rhinosinusitis is a common condition affecting 5–12% of the general population worldwide. In a limited number of cases, the disease is recalcitrant to medical and surgical interventions, causing a major impact on physical, social and emotional well-being and increasing pressure on healthcare systems. Biofilm formation and dysbiosis caused by Staphylococcus aureus and Pseudomonas aeruginosa play a role in the pathogenesis of recalcitrant chronic rhinosinusitis. In these cases, a promising treatment alternative is the application of bacteriophages, which are viruses that infect and lyse bacteria. In this review, we appraise the evidence for the use of bacteriophages in the treatment of recalcitrant chronic rhinosinusitis. Additionally, (dis)advantages of bacteriophages and considerations for implementation of phage therapy in otorhinolaryngology practice will be discussed.

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Biofilm Formation in Xanthomonas arboricola pv. pruni: Structure and Development

Agronomy ◽

10.3390/agronomy11030546 ◽

2021 ◽

Vol 11 (3) ◽

pp. 546

Author(s):

Pilar Sabuquillo ◽

Jaime Cubero

Keyword(s):

Biofilm Formation ◽

Extracellular Polymeric Substances ◽

Environmental Changes ◽

Nutrient Content ◽

Infection Process ◽

Bacterial Attachment ◽

Key Factors ◽

Cell Structures ◽

Microscopy Techniques

Xanthomonasarboricola pv. pruni (Xap) causes bacterial spot of stone fruit and almond, an important plant disease with a high economic impact. Biofilm formation is one of the mechanisms that microbial communities use to adapt to environmental changes and to survive and colonize plants. Herein, biofilm formation by Xap was analyzed on abiotic and biotic surfaces using different microscopy techniques which allowed characterization of the different biofilm stages compared to the planktonic condition. All Xap strains assayed were able to form real biofilms creating organized structures comprised by viable cells. Xap in biofilms differentiated from free-living bacteria forming complex matrix-encased multicellular structures which become surrounded by a network of extracellular polymeric substances (EPS). Moreover, nutrient content of the environment and bacterial growth have been shown as key factors for biofilm formation and its development. Besides, this is the first work where different cell structures involved in bacterial attachment and aggregation have been identified during Xap biofilm progression. Our findings provide insights regarding different aspects of the biofilm formation of Xap which improve our understanding of the bacterial infection process occurred in Prunus spp and that may help in future disease control approaches.

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