Protocol for using negative pressure isolator systems to study BSL-2 organisms in gnotobiotic murine models

Pavao, Aidan; Graham, Madeline; Trofimuk, Olivia; Delaney, Mary L.; Yeliseyev, Vladimir; Bry, Lynn

doi:10.1016/j.xpro.2022.101211

Cited by 2 publications

(4 citation statements)

References 2 publications

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“…C. difficile strain ATCC 43255 (ATCC, Manassas, VA) spores were prepared as described ( 19 ). C. scindens strain ATCC 35704 (ATCC, Manassas, VA) cultures were grown in brain heart infusion broth (3.7%) supplemented with hemin (0.0005%) and vitamin K (0.0001%) in a Coy anaerobic chamber (Coy Labs, Grass Lake, MI) in an atmosphere consisting of 80% N 2 , 10% H 2 , 10% CO 2 at 37°.…”

Section: Methodsmentioning

confidence: 99%

“…Mice were challenged with 1×10 3 C. difficile spores by oral gavage. Progression of disease was assessed via a modified body condition scoring system ( 19 ). Mice were sacrificed at a BCS of 2- (emaciation, lack of ambulation, anal prolapse) or at the following defined timepoints: 7 days of C. scindens colonization or germ-free controls, 16, 20, 24 h, or 14 days post- C. difficile challenge.…”

Section: Methodsmentioning

confidence: 99%

“…Bacterial strains and culture conditions C. difficile strain ATCC 43255 (ATCC, Manassas, VA) spores were prepared as described (19).…”

Section: Methodsmentioning

confidence: 99%

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Clostridium scindens colonization of gnotobiotic mice promotes a chronic unresolving infection with Clostridioides difficile

Graham

DiBenedetto

Delaney

et al. 2022

Preprint

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The commensal Clostridium scindens has been regarded as a promising bacteriotherapeutic against Clostridioides difficile infection due to its ability to consume host factors that can promote C. difficile growth, and its production of the antimicrobial compound 1-acetyl-β-carboline. We investigated C. scindens' protective effects against C. difficile using defined colonization studies in gnotobiotic mice. Mice infected with C. difficile develop lethal infection within 48 hours. In contrast, 88% of mice pre-colonized with C. scindens survived acute infection with delayed C. difficile colonization, lower biomass, and toxin B levels at 24 hours after infection. However, two weeks post-challenge, surviving mice showed comparable levels of cecal C. difficile vegetative and spore biomass and toxin B, as seen during acute infection. After two weeks, co-colonized mice exhibited mucosal colonic hyperplasia with focal pseudomembranes , modeling a chronic and recurrent infection state. Our findings illustrate how the commensal microbiota can modulate host and pathogen interactions leading to chonic C. difficile carriage and infection.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Clostridium scindens colonization of gnotobiotic mice promotes a chronic unresolving infection with Clostridioides difficile

Graham

DiBenedetto

Delaney

et al. 2022

Preprint

Self Cite

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“…Working with these models requires meticulous protocols and good experimental controls, as well as trained personnel. Germ-free or gnotobiotic mice are bred and housed in isolators, either flexible (under mandatory positive pressure) or rigid, the latter being suitable to work under positive or negative pressure [63]. Isolators are ventilated with HEPA-filtered sterile air (High-Efficiency Particulate Air), and food, water and other supplies must be appropriately sterilized using autoclave, irradiation or treatment with disinfectants [64,65].…”

Section: Advantages and Disadvantages Of Gnotobiotic Mouse Modelsmentioning

confidence: 99%

The state of play of rodent models for the study of Clostridioides difficile infection

Brosse,

Coullon,

Janoir

et al. 2024

Journal of Medical Microbiology

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Clostridioides difficile is the most common cause of nosocomial antibiotic-associated diarrhoea and is responsible for a spectrum of diseases characterized by high levels of recurrence and morbidity. In some cases, complications can lead to death. Currently, several types of animal models have been developed to study various aspects of C. difficile infection (CDI), such as colonization, virulence, transmission and recurrence. These models have also been used to test the role of environmental conditions, such as diet, age and microbiome that modulate infection outcome, and to evaluate several therapeutic strategies. Different rodent models have been used successfully, such as the hamster model and the gnotobiotic and conventional mouse models. These models can be applied to study either the initial CDI infectious process or recurrences. The applications of existing rodent models and their advantages and disadvantages are discussed here.

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Protocol for using negative pressure isolator systems to study BSL-2 organisms in gnotobiotic murine models

Cited by 2 publications

References 2 publications

Clostridium scindens colonization of gnotobiotic mice promotes a chronic unresolving infection with Clostridioides difficile

Clostridium scindens colonization of gnotobiotic mice promotes a chronic unresolving infection with Clostridioides difficile

The state of play of rodent models for the study of Clostridioides difficile infection

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