• Clostridioides difficile
• Clostridium botulinum
• Clostridium perfringens
• Clostridium tetani
• Antibiotic resistance
• Postbiotics

• Clostridium Infections/drug therapy
• Clostridium Infections/microbiology
• Clostridium Infections/therapy
• Probiotics/pharmacology
• Humans
• Anti-Bacterial Agents/pharmacology
• Animals

• Clostridioides difficile
• antimicrobial
• biocorrosion
• biofilm kill
• biofilm prevention
• electrochemical test kit

• Clostridioides difficile infection
• cationic peptidomimetic
• cephamycins
• membrane disruption
• sporulation
• therapeutics
• virulence factors
• β-lactam

• 1145760 National Health and Medical Research Council of Australia

• Clostridioides difficile
• genetic epidemiology
• hospital-acquired infection
• whole-genome sequencing

• Animals
• Humans
• Clostridioides difficile/genetics
• Clostridium Infections/epidemiology
• Genomics
• Cross Infection/epidemiology
• Australia

• FT120100779 Department of Education and Training | Australian Research Council (ARC)

• AgAu
• AgCl
• Clostridioides difficile infection
• peroxide
• spore

• Ministry of Science and Technology, Taiwan

• Listeria monocytogenes
• classical methods
• genotyping
• identification
• molecular methods

Clostridioides difficile (previously Clostridium difficile) causes life-threatening gut infections. The central metabolism of the bacterium is strongly influencing toxin production and consequently the infection progress. In this context, the composition and potential origin of the volatile metabolome was investigated, showing a large number of sulfur-containing volatile metabolites. Gas chromatography/mass spectrometry (GC/MS)-based headspace analyses of growing C. difficile 630Δerm cultures identified 105 mainly sulfur-containing compounds responsible of the typical C. difficile odor. Major components were identified to be 2-methyl-1-propanol, 2-methyl-1-propanethiol, 2-methyl-1-butanethiol, 4-methyl-1-pentanethiol, and as well as their disulfides. Structurally identified were 64 sulfur containing volatiles. In order to determine their biosynthetic origin, the concentrations of the sulfur-containing amino acids methionine and cysteine were varied in the growth medium. The changes observed in the volatile metabolome profile indicated that cysteine plays an essential role in the formation of the sulfur-containing volatiles. We propose that disulfides are derived from cysteine via formation of cystathionine analogs, which lead to corresponding thiols. These thiols may then be oxidized to disulfides. Moreover, methionine may contribute to the formation of short-chain disulfides through integration of methanethiol into the disulfide biosynthesis. In summary, the causative agents of the typical C. difficile odor were identified and first hypotheses for their biosynthesis were proposed.

• Clostridium difficile
• immunoinformatics
• molecular docking
• molecular dynamics simulation
• multi-epitope vaccine

• Clostridioides difficile
• Diarrhea
• Epitopes, B-Lymphocyte
• Epitopes, T-Lymphocyte
• Humans
• Molecular Docking Simulation
• Vaccines, Subunit

• Natural Science Foundation of Hunan Province

• Antibiotic
• Clostridioides difficile
• Hospital
• Infection control
• Time series data

The capability of some representatives of Clostridium spp. and Bacillus spp. genera to form spores in extreme external conditions long ago became a subject of medico-biological investigations. Bacterial spores represent dormant cellular forms of gram-positive bacteria possessing a high potential of stability and the capability to endure extreme conditions of their habitat. Owing to these properties, bacterial spores are recognized as the most stable systems on the planet, and spore-forming microorganisms became widely spread in various ecosystems.

Spore-forming bacteria have been attracted increased interest for years due to their epidemiological danger. Bacterial spores may be in the quiescent state for dozens or hundreds of years but after they appear in the favorable conditions of a human or animal organism, they turn into vegetative forms causing an infectious process. The greatest threat among the pathogenic spore-forming bacteria is posed by the causative agents of anthrax (B. anthracis), food toxicoinfection (B. cereus), pseudomembranous colitis (C. difficile), botulism (C. botulinum), gas gangrene (C. perfringens).

For the effective prevention of severe infectious diseases first of all it is necessary to study the molecular structure of bacterial spores and the biochemical mechanisms of sporulation and to develop innovative methods of detection and disinfection of dormant cells. There is another side of the problem: the necessity to investigate exo- and endospores from the standpoint of obtaining similar artificially synthesized models in order to use them in the latest medical technologies for the development of thermostable vaccines, delivery of biologically active substances to the tissues and intracellular structures. In recent years, bacterial spores have become an interesting object for the exploration from the point of view of a new paradigm of unicellular microbiology in order to study microbial heterogeneity by means of the modern analytical tools.

• Raman spectroscopy
• atomic-force microscopy
• bacterial spores
• biotechnologies
• endospores
• exospores
• plasma chemical techniques.
• sporulation

Spore-forming bacteria are a major concern for the food industry as they cause both spoilage and food safety issues. Moreover, as they are more resistant than vegetative cells, their removal from the food processing environment may be difficult to achieve. This study investigated the efficacy of the ten most commonly used disinfectant agents (assigned 1–10), used at the recommended concentrations in the meat industry, for their ability to eliminate Clostridium sporogenes and Clostridioides difficile spores. Test-tube based suspension assays suggested that disinfectants 2 (10% v/v preparation of a mixture of hydrogen peroxide (10–30%), acetic acid (1–10%) and peracetic acid (1–10%)), 7 (4% w/v preparation of a mixture of peroxymonosulphate (30–50%), sulphamic acid (1–10%) and troclosene sodium (1–10%)) and 10 (2% v/v preparation of a mixture of glutaraldehyde (10–30%), benzalkonium chloride (1–10%)) were the most effective formulations. D-values for these ranged from 2.1 to 8.4 min at 20 °C for the target spores. Based on these findings, it is recommended that these disinfectants are used to control Clostridium spores in the meat plant environment.

• Clostridioides difficile
• Clostridium sporogenes
• D values
• disinfectants
• spores
• sporicidal agents

• C. difficile infection
• chimeric antigen
• heat labile E. coli enterotoxin B subunit
• molecular farming
• oral immunization

• Administration, Oral
• Animals
• Antibodies, Bacterial/blood
• Antigens, Bacterial/genetics
• Antigens, Bacterial/metabolism
• Clostridioides difficile/genetics
• Enterotoxins/genetics
• Escherichia coli Proteins/genetics
• Immunoglobulin G/blood
• Mice
• Mice, Inbred BALB C
• Molecular Farming
• Plants, Genetically Modified/genetics
• Plants, Genetically Modified/metabolism
• Spores, Bacterial/genetics
• Nicotiana/genetics
• Nicotiana/metabolism
• Vaccines, Edible/genetics
• Vaccines, Edible/immunology
• Vaccines, Edible/metabolism

• Clostridioides difficile
• antimicrobials
• environment
• hemodialysis
• infections
• transmission

• Clostridioides
• Clostridioides difficile
• Clostridium Infections/epidemiology
• Clostridium Infections/prevention & control
• Humans
• Outpatients
• Renal Dialysis/adverse effects

• CC999999 Intramural CDC HHS

Clostridioides difficile is a Gram-positive, spore-forming bacterium that causes a severe intestinal infection. Spores of this pathogen enter in the human body through the oral route, interact with intestinal epithelial cells and persist in the gut. Once germinated, the vegetative cells colonize the intestine and produce toxins that enhance an immune response that perpetuate the disease. Therefore, spores are major players of the infection and ideal targets for new therapies. In this context, spore surface proteins of C. difficile, are potential antigens for the development of vaccines targeting C. difficile spores. Here, we report that the C-terminal domain of the spore surface protein BclA3, BclA3_CTD, was identified as an antigenic epitope, over-produced in Escherichia coli and tested as an immunogen in mice. To increase antigen stability and efficiency, BclA3_CTD was also exposed on the surface of B. subtilis spores, a mucosal vaccine delivery system. In the experimental conditions used in this study, free BclA3_CTD induced antibody production in mice and attenuated some C. difficile infection symptoms after a challenge with the pathogen, while the spore-displayed antigen resulted less effective. Although dose regimen and immunization routes need to be optimized, our results suggest BclA3_CTD as a potentially effective antigen to develop a new vaccination strategy targeting C. difficile spores.

• Bacillus subtilis
• Clostridium difficile
• exosporium proteins
• gastrointestinal infection
• immune response
• mucosal vaccine
• recombinant spores

• Bacterial spores
• Flow cytometry
• Light scatter (FSC/SSC)
• Pseudoclostridium thermosuccinogenes
• cFDA/PI dyes

• Clostridioides (Clostridium) difficile
• Enterococcus faecalis
• intestinal cell
• probiotics
• spores

Clostridioides difficile is a leading cause of health care-associated infections. Although significant progress has been made in the understanding of its genome, the epigenome of C. difficile and its functional impact has not been systematically explored. Here, we performed a comprehensive DNA methylome analysis of C. difficile using 36 human isolates and observed great epigenomic diversity. We discovered an orphan DNA methyltransferase with a well-defined specificity whose corresponding gene is highly conserved across our dataset and in all ∼300 global C. difficile genomes examined. Inactivation of the methyltransferase gene negatively impacted sporulation, a key step in C. difficile disease transmission, consistently supported by multi-omics data, genetic experiments, and a mouse colonization model. Further experimental and transcriptomic analysis also suggested that epigenetic regulation is associated with cell length, biofilm formation, and host colonization. These findings provide a unique epigenetic dimension to characterize medically relevant biological processes in this critical pathogen. This work also provides a set of methods for comparative epigenomics and integrative analysis, which we expect to be broadly applicable to bacterial epigenomics studies.

• Clostridium difficile
• biocide
• public health
• spores
• surfaces
• survival
• transmission

• Clostridioides difficile
• Environmental contamination
• Polywipes
• Surface sampling
• Transmission

• Electronic health records and systems
• biosurveillance and case reporting
• data visualization

Spore-forming pathogenic bacteria, such as Clostridium difficile, are associated with nosocomial infection, leading to the increased use of sporicidal disinfectants, which impacts socioeconomic costs. However, C. difficile can be prevented using microorganisms such as Bacillus amyloliquefaciens, a prophylactic agent that has been proven to be effective against it in recent tests or it can be controlled by sporicidal disinfectants. These disinfectants against spores should be evaluated according to a known and recommended standard. Unfortunately, some newly manufactured disinfectants like Bioxy products have not yet been tested. ASTM E2197-11 is a standard test that uses stainless steel disks (1 cm in diameter) as carriers, and the performance of the test formulation is calculated by comparing the number of viable test organisms to that on the control carriers. Surface tests are preferable for evaluating disinfectants with sporicidal effects on hard surfaces. This study applies improved methods, based on the ASTM E2197-11 standard, for evaluating and comparing the sporicidal efficacies of several disinfectants against spores of C. difficile and B. amyloliquefaciens, which are used as the test organisms. With the improved method, all spores were recovered through vortexing and membrane filtration. The results show that chlorine-based products are effective in 5 min and Bioxy products at 5% w/v are effective in 10 min. Although Bioxy products may take longer to prove their effectiveness, their non-harmful effects to hospital surfaces and people have been well established in the literature.

• Bacillus amyloliquefaciens
• Clostridium difficile
• disinfectant
• spores
• sporicidal

Unrelated spore-forming bacteria share unique characteristics stemming from the presence of highly resistant endospores, leading to similar challenges in health and disease. These characteristics are related to the presence of these highly transmissible spores, which are commonly spread within the environment and are implicated in host-to-host transmission. In humans, spore-forming bacteria contribute to a variety of pathological processes that share similar characteristics, including persistence, chronicity, relapses and the maintenance of the resistome. We first outline the necessity of characterizing the totality of the spore-forming bacteria as the sporobiota based on their unique common characteristics. We further propose that the collection of all genes of spore-forming bacteria be known as the sporobiome. Such differentiation is critical for exploring the cross-talk between the sporobiota and other members of the gut microbiota, and will allow for a better understanding of the implications of the sporobiota and sporobiome in a variety of pathologies and the spread of antibiotic resistance.

• Bacteriophages
• Clostridium botulinum
• Clostridium difficile
• Clostridium novyi
• Clostridium perfringens
• Lysogenic conversion
• Prophages
• Toxins
• Virulence

• Clostridioides difficile
• Clostridium difficile
• Fermentation profile
• Metabolism
• Stickland reaction

• Bacterial contamination
• Disinfection
• Healthcare-associated infection
• Patient safety
• Ultrasound

• Bacteria/isolation & purification
• Equipment Contamination/statistics & numerical data
• Gels
• Infection Control
• Polymerase Chain Reaction
• Risk
• Risk Factors
• Ultrasonography/instrumentation

Clostridium difficile is the causative agent of the most frequently reported nosocomial diarrhea worldwide. The high incidence of recurrent infection is the main clinical challenge of C. difficile infections (CDI). Formation of C. difficile spores of the epidemic strain R20291 has been shown to be essential for recurrent infection and transmission of the disease in a mouse model. However, the underlying mechanisms of how these spores persist in the colonic environment remains unclear. In this work, we characterized the adherence properties of epidemic R20291 spores to components of the intestinal mucosa, and we assessed the role of the exosporium integrity in the adherence properties by using cdeC mutant spores with a defective exosporium layer. Our results showed that spores and vegetative cells of the epidemic R20291 strain adhered at high levels to monolayers of Caco-2 cells and mucin. Transmission electron micrographs of Caco-2 cells demonstrated that the hair-like projections on the surface of R20291 spores are in close proximity with the plasma membrane and microvilli of undifferentiated and differentiated monolayers of Caco-2 cells. Competitive-binding assay in differentiated Caco-2 cells suggests that spore-adherence is mediated by specific binding sites. By using spores of a cdeC mutant we demonstrated that the integrity of the exosporium layer determines the affinity of adherence of C. difficile spores to Caco-2 cells and mucin. Binding of fibronectin and vitronectin to the spore surface was concentration-dependent, and depending on the concentration, spore-adherence to Caco-2 cells was enhanced. In the presence of an aberrantly-assembled exosporium (cdeC spores), binding of fibronectin, but not vitronectin, was increased. Notably, independent of the exosporium integrity, only a fraction of the spores had fibronectin and vitronectin molecules binding to their surface. Collectively, these results demonstrate that the integrity of the exosporium layer of strain R20291 contributes to selective spore adherence to components of the intestinal mucosa.

• BclA
• C. difficile spores
• CdeC
• exosporium
• spore adherence

Clostridium difficile infection (CDI) is the most common nosocomial infection in the United States and is associated with a high mortality. One quarter of patients treated for CDI have at least one recurrence. Spore persistence, impaired host immune response and alteration in the gastrointestinal microbiome due to antibiotic use are factors in recurrent disease. We review the etiology of recurrent CDI and best approaches to management including fecal microbiota transplantation.

• Clostridium difficile infection
• Epidemiology
• Outcomes
• Treatment
• Fecal microbiota transplantation

• Clostridium difficile
• antibiotic resistance
• antibiotic-associated diarrhea
• microbiome
• microbiota
• mobile genetic elements

• BclA1
• CdeC
• CdeM
• Clostridium difficile
• CotA
• Exosporium
• Spores
• Vaccine

• Animals
• Bacterial Proteins/immunology
• Bacterial Vaccines/immunology
• Clostridioides difficile/immunology
• Female
• Male
• Mesocricetus
• Mice
• Mice, Inbred C57BL
• Spores, Bacterial/immunology
• Vaccines, Synthetic/immunology

• AI109526 NIAID NIH HHS
• DK101870 NIDDK NIH HHS
• R21 AI113470 NIAID NIH HHS
• R56 AI095256 NIAID NIH HHS
• R03 DK101870 NIDDK NIH HHS
• K01DK092352 NIDDK NIH HHS
• K01 DK092352 NIDDK NIH HHS
• R01 AI095256 NIAID NIH HHS
• K01 DK087763 NIDDK NIH HHS
• DK087763 NIDDK NIH HHS
• R56 AI109526 NIAID NIH HHS
• R01AI095256-01A1 NIAID NIH HHS

• AAD, antibiotic-associated diarrhea
• CDI, Clostridium difficile infection
• CPD, cysteine proteinase domain
• GTD, glucosyltransferase domain
• HuMabs, human monoclonal antibodies
• IVIG, intravenous immunoglobulin
• RBD, receptor binding domain
• SLP, surface-layer protein
• TMD, transmembrane domain
• bacterial toxins
• clostridium difficile infection (CDI)
• immunotherapy
• mAb, monoclonal antibody
• monoclonal antibody
• vaccine

• Clostridium difficile
• Host response
• Microbiota
• Spore
• Virulence factors

• Clostridium
• Cortex hydrolysis
• Germinant receptors
• Germination
• Spores