Microbiological growth media and methods of using the same
Inventors
SHARA, Kate • SCHWEDOCK, Julie • VOGEL, Sommer
Assignees
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Publication Number
US-11732237-B2
Publication Date
2023-08-22
Expiration Date
Abstract
The present invention features general-purpose microbiological growth media capable of supporting growth of microorganisms on membranes. The media contain casein digest, soybean digest, animal tissue digest, yeast extract, dextrose, a phosphate buffer, hemin, and L-cystine. The invention features an all-purpose microbiological growth media that can support the growth of anaerobes, molds, injured spores, and general aerobic bacteria to a greater extent than other media.
Core Innovation
The invention relates to an all-purpose microbiological growth medium for membrane-based sterility testing. The medium supports growth of a broad range of organisms, including anaerobes, molds, injured spores, and aerobic bacteria, using a defined nutrient composition.
The composition comprises casein digest, soybean digest, animal tissue digest, yeast extract, dextrose, a phosphate buffer, hemin, and L-cystine, while explicitly requiring that the composition does not contain added sodium. The medium further supports defined formulation constraints including pH about 7.3 ± 0.5 and specified buffer capacity behavior.
The document also describes that the medium may be provided in solid/powder, liquid, or gel forms and may include optional components such as gelling agents, including agar, gellan, sodium alginate, xanthan gum, and others, and surfactants, including polysorbate 20. It additionally describes culturing approaches compatible with permeable membranes, including contacting growth using opposite sides of permeable membranes in the membrane-based sterility testing context.
Claims Coverage
The claims are centered on one independent composition claim defining a specific nutrient formulation, with dependent claims refining composition parameters and extending to a culturing method using the defined composition. The main inventive features include the defined digest-and-cofactor nutrient set and the explicit exclusion of added sodium, together with quantitative constraints and optional formulation elements.
Defined nutrient composition without added sodium
A composition comprising casein digest, soybean digest, animal tissue digest, yeast extract, dextrose, a phosphate buffer, hemin, and L-cystine, wherein said composition does not contain added sodium.
Casein digest amount excluding phosphate buffer
Excluding said phosphate buffer from the total mass of said composition, said composition comprises between about 1 g/kg and about 500 g/kg of casein digest.
Quantified pH range
The composition is defined to have a pH of 7.3 ± 0.5.
Phosphate buffer defined by buffer capacity
The claim covers a composition that includes phosphate buffer in an amount sufficient to provide a buffer capacity of from 0.1 mmol/(pH unit) to 100 mmol/(pH unit) upon dissolution in an aqueous medium.
Disinfectant neutralizer selection
The composition includes a neutralizer selected from histidine, thiosulfate, polysorbate 80, and/or lecithin.
Culturing cells using the defined composition under growth-supportive conditions
The claim covers culturing a population of cells by contacting them with the composition under growth-supportive conditions.
Overall, the independent claim defines a nutrient medium composition with specific digests and supplements, including hemin and L-cystine, and requires the absence of added sodium. The dependent claims further delimit key formulation parameters such as casein digest concentration, pH, and phosphate buffer buffer capacity, optionally introduce specified neutralizers, and cover a culturing approach using the defined composition under growth-supportive conditions.
Stated Advantages
Improved recoveries versus standard media and Schaedler formulations, including performance with stressed microorganisms such as bleach- and oxidatively stressed Bacillus subtilis.
Supports faster growth for Propionibacterium acnes in anaerobic cassettes versus FTM as described.
Documented Applications
Membrane-based sterility testing using a microbiological growth medium that supports growth across a broad range of organisms.
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