Single cell analysis of nutrient regulation of Clostridioides (Clostridium) difficile motility

Courson, David S.; Pokhrel, Astha; Scott, Cody B.; Madrill, Melissa; Rinehold, Alden J.; Tamayo, Rita; Cheney, Richard E.; Purcell, Erin B.

doi:10.1016/j.anaerobe.2019.102080

Cited by 12 publications

(13 citation statements)

References 58 publications

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“…As C. difficile displays motility in agarose ( Baban et al, 2013 ; Courson et al, 2019 ), we wondered if altering the matrix elasticity of the medium has an influence on the motility phenotype. High molecular weight polymers such as polyvinylpyrrolidone (PVP), which increase the matrix elasticity, were described to enhance motility in various bacteria in a concentration dependent manner ( Schneider and Doetsch, 1974 ; Greenberg and Canale-Parola, 1977 ).…”

Section: Resultsmentioning

confidence: 99%

“…Flagellar motility and chemotaxis are important for successful colonisation and virulence of many gastrointestinal pathogens, for example Campylobacter jejuni , Salmonella enterica Serovar Typhimurium, Helicobacter pylori , and Vibrio chlolerae ( Boin et al, 2004 ; Stecher et al, 2004 ; Lertsethtakarn et al, 2011 ; Korolik, 2019 ). Most C. difficile strains produce peritrichous flagella, which can mediate swimming motility in soft-agar based assays ( Twine et al, 2009 ; Baban et al, 2013 ; Courson et al, 2019 ). The contribution of flagellar motility for the pathogenesis in mice was studied with the aid of C. difficile flagellar mutants, which were found to be reduced in their colonisation efficiency ( Baban et al, 2013 ; Batah et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

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Clostridioides difficile Single Cell Swimming Strategy: A Novel Motility Pattern Regulated by Viscoelastic Properties of the Environment

et al. 2021

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Flagellar motility is important for the pathogenesis of many intestinal pathogens, allowing bacteria to move to their preferred ecological niche. Clostridioides difficile is currently the major cause for bacterial health care-associated intestinal infections in the western world. Most clinical strains produce peritrichous flagella and are motile in soft-agar. However, little knowledge exists on the C. difficile swimming behaviour and its regulation at the level of individual cells. We report here on the swimming strategy of C. difficile at the single cell level and its dependency on environmental parameters. A comprehensive analysis of motility parameters from several thousand bacteria was achieved with the aid of a recently developed bacterial tracking programme. C. difficile motility was found to be strongly dependent on the matrix elasticity of the medium. Long run phases of all four motile C. difficile clades were only observed in the presence of high molecular weight molecules such as polyvinylpyrrolidone (PVP) and mucin, which suggests an adaptation of the motility apparatus to the mucin-rich intestinal environment. Increasing mucin or PVP concentrations lead to longer and straighter runs with increased travelled distance per run and fewer turnarounds that result in a higher net displacement of the bacteria. The observed C. difficile swimming pattern under these conditions is characterised by bidirectional, alternating back and forth run phases, interrupted by a short stop without an apparent reorientation or tumbling phase. This motility type was not described before for peritrichous bacteria and is more similar to some previously described polar monotrichous bacteria.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Clostridioides difficile Single Cell Swimming Strategy: A Novel Motility Pattern Regulated by Viscoelastic Properties of the Environment

et al. 2021

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“…As C. difficile exhibits green autoflouresence, the red TAMRA labeling was distinct from any intrinsic signal produced by the cells [62]. Exponential-phase cells and peptides were mixed and sealed within microscopy chambers in an anaerobic chamber and then transported to the microscope, resulting in a 6-min delay between the onset of peptide exposure and the first image [63]. Mean fluorescence intensity within cells was stable over the course of 1 h of monitoring, indicating that peptide incorporation into cells occurs within the first few min of exposure (Figure 1A,B).…”

Section: Resultsmentioning

confidence: 99%

“…1.49), pco.edge 4.2 LT sCMOS camera, and SOLA SE II 365 Light Engine as well as complementary DIC components (Nikon Instruments Inc, Melville, NY, USA). Mid-logarithmic phase cells and peptides at the indicated concentration were mixed inside the anaerobic chamber and injected into home-built anaerobic rose-type imaging chambers as previously described [63]. Imaging chambers were removed from the anaerobic chamber and placed on the microscope.…”

Section: Methodsmentioning

confidence: 99%

How Oxygen Availability Affects the Antimicrobial Efficacy of Host Defense Peptides: Lessons Learned from Studying the Copper-Binding Peptides Piscidins 1 and 3

Oludiran

Courson

Stuart

et al. 2019

IJMS

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The development of new therapeutic options against Clostridioides difficile (C. difficile) infection is a critical public health concern, as the causative bacterium is highly resistant to multiple classes of antibiotics. Antimicrobial host-defense peptides (HDPs) are highly effective at simultaneously modulating the immune system function and directly killing bacteria through membrane disruption and oxidative damage. The copper-binding HDPs piscidin 1 and piscidin 3 have previously shown potent antimicrobial activity against a number of Gram-negative and Gram-positive bacterial species but have never been investigated in an anaerobic environment. Synergy between piscidins and metal ions increases bacterial killing aerobically. Here, we performed growth inhibition and time-kill assays against C. difficile showing that both piscidins suppress proliferation of C. difficile by killing bacterial cells. Microscopy experiments show that the peptides accumulate at sites of membrane curvature. We find that both piscidins are effective against epidemic C. difficile strains that are highly resistant to other stresses. Notably, copper does not enhance piscidin activity against C. difficile. Thus, while antimicrobial activity of piscidin peptides is conserved in aerobic and anaerobic settings, the peptide–copper interaction depends on environmental oxygen to achieve its maximum potency. The development of pharmaceuticals from HDPs such as piscidin will necessitate consideration of oxygen levels in the targeted tissue.

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“…Soft agar can, up to some extent, mimic physical, chemical, and nutritional conditions inside and outside the host [ 29 , 72 , 73 ]. Furthermore, chambers can mimic environments such as xylem vessels [ 74 ], enabling the discovery that Xylella fastidiosa migrates against the flow via twitching motility, and anaerobiosis, allowing researchers to prove that Clostridioides difficile modulates its swimming speed in the presence of a metabolite related to its host colonization [ 75 ]. Likewise, vertical diffusion chambers (VDC) were used to study the role of motility in Campylobacter jejuni invasion of epithelial cells [ 76 ].…”

Section: Study Of Bacterial Motility In Bacterial–host Interactionsmentioning

confidence: 99%

Methods to Evaluate Bacterial Motility and Its Role in Bacterial–Host Interactions

et al. 2022

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Bacterial motility is a widespread characteristic that can provide several advantages for the cell, allowing it to move towards more favorable conditions and enabling host-associated processes such as colonization. There are different bacterial motility types, and their expression is highly regulated by the environmental conditions. Because of this, methods for studying motility under realistic experimental conditions are required. A wide variety of approaches have been developed to study bacterial motility. Here, we present the most common techniques and recent advances and discuss their strengths as well as their limitations. We classify them as macroscopic or microscopic and highlight the advantages of three-dimensional imaging in microscopic approaches. Lastly, we discuss methods suited for studying motility in bacterial–host interactions, including the use of the zebrafish model.

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Single cell analysis of nutrient regulation of Clostridioides (Clostridium) difficile motility

Cited by 12 publications

References 58 publications

Clostridioides difficile Single Cell Swimming Strategy: A Novel Motility Pattern Regulated by Viscoelastic Properties of the Environment

Clostridioides difficile Single Cell Swimming Strategy: A Novel Motility Pattern Regulated by Viscoelastic Properties of the Environment

How Oxygen Availability Affects the Antimicrobial Efficacy of Host Defense Peptides: Lessons Learned from Studying the Copper-Binding Peptides Piscidins 1 and 3

Methods to Evaluate Bacterial Motility and Its Role in Bacterial–Host Interactions

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