Improved microbial growth media
AssigneeBecton, Dickinson and Company
InventorsGretta L. Koontz, Stephanie Ann Frey, Kristina Anne Robinson
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Abstract
The compositions, apparatuses, and methods described herein relate to the discovery that the time to detection of yeast and microorganism contaminants in microbial growth media can be improved by controlling the concentration of iron and copper.
Claims63
A microbial growth media providing an increased growth rate of Candida, the microbial growth media comprising:
- a concentration of iron of about 1 to about 400 μM;
- and
- optionally, a concentration of copper of up to about 700 μM;
- wherein the growth rate of Candida cultured in the microbial growth media is increased as compared to the same microbial growth media in the absence of the iron and copper.
5 to 3.5 g/L D-trehalose, 0.5 to 1 g/L L-glutamic acid, 0.0001 to 0.0012 g/L L-cysteine, 0.5 to 60 g/L TSB, 0.5 to 1.5 g/L SPS, 0.0004 to 0.0012 g/L menadione, 0.005 to 0.025 g/L Pyridoxal HCl, 0.001 to 0.1 g/L ferulic acid, 0.5 to 3 g/L sodium hydroxide, 0.01 to 0.25 g/L ascorbic acid, and 0.001 to 0.1 g/L hemin, in water.
The microbial growth media of any one of the preceding claims, wherein the reduced TTD of Candida cultured in the microbial growth media is determined by:
- amending culture containers comprising the microbial growth media with and without the iron and copper with 0.5, 3, or 10 ml of blood;
- inoculating the culture containers with 0.1 mL of a culture of Candida to provide 10-100 CFU of Candida per container;
- culturing the Candida at about 30° C. to 37° C. for up to 120 hours;
- monitoring the culture containers for a signal indicative of the presence of Candida;
- and
- determining the TTD of the Candida in the microbial growth media with and without the iron and copper.
The microbial growth media of any one of the preceding claims, wherein the Candida is Candida albicans or Candida glabrata.
The microbial growth media of any one of the preceding claims, wherein the microbial growth media comprises, or comprises about, 0, 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1.0 g/L yeast nitrogen base, or a range defined by any two of the preceding values.
The microbial growth media of any one of the preceding claims, wherein the microbial growth media comprises less than 0.01% w/v yeast nitrogen base.
The microbial growth media of any one of the preceding claims, wherein the yeast nitrogen base does not comprise histidine, methionine, and tryptophan.
The microbial growth media of any one of the preceding claims, wherein the microbial growth media comprises, or comprises about, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 28, 50, 75, 100, 125, 150, 170, 175, 200, 225, 250, 275, 300, 325, 350, or 400 μM iron, or a range defined by any two of the preceding values, optionally 1 to 400 μM, 1 to 350 μM, 1 to 200 μM, 1 to 100 μM, 1 to 50 μM, 2 to 400 μM, 2 to 350 μM, 25 to 400 μM, 25 to 325 μM, 25 to 200μM, 25 to 100 μM, or 25 to 75 μM.
The microbial growth media of any one of the preceding claims, wherein concentration of iron is about 28 μM.
The microbial growth media of any one of the preceding claims, wherein concentration of iron is about 100 μM.
The microbial growth media of any one of the preceding claims, wherein concentration of iron is about 170 μM.
The microbial growth media of any one of the preceding claims, wherein the iron is ferric iron.
The microbial growth media of any one of the preceding claims, wherein the iron is provided as ferric ammonium citrate, ferric chloride, ferric sulfate, ferric nitrate.
The microbial growth media of any one of the preceding claims, wherein the microbial growth media comprises, or comprises about, 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 150, 200, 300, 400, 500, 600, 650, or 700 μM copper, or a range defined by any two of the preceding values, optionally 0 to 700 μM, 0 to 650 μM, 0 to 500 μM, 0 to 300 μM, 0 to 100 μM, 0 to 50 μM, 0 to 25 μM, 0.1 to 100 μM, 0.1 to 50 μM, 0.1 to 25 μM, 0.5 to 650 μM, 0.5 to 300 μM, 0.5 to 100 μM, 0.5 to 50 μM, or 0.5 to 25 μM.
The microbial growth media of any one of the preceding claims, wherein the concentration of copper is about 3 μM.
The microbial growth media of any one of the preceding claims, wherein the copper is cupric copper.
The microbial growth media of any one of the preceding claims, wherein the copper is provided as cupric sulfate, cupric chloride, cupric nitrate, cupric bromide, cupric chlorate, and/or copper(II) gluconate.
The microbial growth media of any one of the preceding claims, wherein the microbial growth media is an aqueous liquid growth media further comprising sucrose, dextrose, D-Trehalose, yeast extract, L-glutamic acid, tryptic soy broth, sodium polyanethole sulfonate, menadione, pyridoxal HCl, ferulic acid, sodium hydroxide, ascorbic acid, L-Cysteine, hemin.
The microbial growth media of any one of the preceding claims, wherein the microbial growth medium comprises, or comprises about, 1.0 to 10.0 g/L yeast extract, 0.15 to 1.5 g/L sucrose, 1.0 to 10.0 g/L dextrose, 1.0 to 10.0 g/L D-trehalose, 0.001 to 1.5 g/L L-glutamic acid, 0.00001 to 0.01 g/L L-cysteine, 0.5 to 15 g/L TSB, 0.001 to 1.5 g/L SPS, 0.00001 to 0.01 g/L menadione, 0.0001 to 1 g/L Pyridoxal HCl, 0.0001 to 1 g/L ferulic acid, 0.1 to 10 g/L sodium hydroxide, 0.001 to 1.0 g/L ascorbic acid, and 0.0001 to 1 g/L hemin, in water.
The microbial growth media of any one of the preceding claims, wherein the microbial growth medium comprises, or comprises about, 0.001 g/L to 0.15 g/L iron, 0.0001 to 0.001 g/L copper, 3.0 to 5.0 g/L yeast extract, 0.15 to 1.5 g/L sucrose, 2.5 to 3.5 g/L dextrose,
The microbial growth media of any one of the preceding claims, wherein the microbial growth media comprises, or comprises about, 0.01 g/L iron;
- 0.001 g/L copper;
- 0.528 g/L sucrose;
- 3.2 g/L dextrose;
- 3.2 g/L D-trehalose;
- 4 g/L yeast extract;
- 0.8 g/L L-glutamic acid;
- 44 g/L TSB;
- 0.8 g/L SPS, 0.0008 g/L menadione, 0.016 g/L Pyridoxal HCl;
- 0.008 g/L ferulic acid;
- 1.225 g/L sodium hydroxide;
- 0.08 g/L ascorbic acid;
- 0.0008 g/L L-cysteine, and 0.008 g/L g hemin.
The microbial growth media of any one of the preceding claims, wherein the microbial growth media comprises, or comprises about, 0.01 g/L iron;
- 0.001 g/L copper;
- 0.33 g/L sucrose;
- 2 g/L dextrose;
- 2 g/L D-trehalose;
- 2.5 g/L yeast extract;
- 0.5 g/L L-glutamic acid;
The microbial growth media of any one of the preceding claims, wherein the microbial growth media is able to support the growth of one or more microbial species selected from the group consisting of Abiotrophia defectiva, Acinetobacter lwoffii, Agreggatibacter actinomycetemcomitans, Aerococcus viridins, Alcaligenes faecalis, Bacillus subtilis, Candida albicans, Candida glabrata, Cardiobacterium hominis, Corynebacterium jeikeium, Cryptococcus neoformans, Eikinella corrodens, Enterobacter cloacae, Enterococcus faecalis, Escherichia coli, Granulicatella adiacens, Haemophilus influenzae, Haemophilus influenzae, Haemophilus influenzae type a, Haemophilus influenzae type b, Haemophilus parainfluenzae, Kingella kingae, Klebsiella pneumonia, Leuconostoc species, Micrococcus luteus, Neisseria gonorrhoeae, Neisseria meningitidis, Pediococcus acidilactici, Proteus mirabilis, Providencia stuartii, Pseudomonas aeruginosa, Rothia mucilaginosa, Saccharomyces cerevisiae, Staphylococcus aureus, Staphylococcus epidermidis, Stenotrophomonas maltophilia, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pneumoniae, Streptococcus pneumoniae, Streptococcus pneumoniae, Streptococcus pyogenes, and Streptococcus sanguinis.
The microbial growth media of any one of the preceding claims, wherein the increase in growth rate of Candida is, is about, is at least, is at least about, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 75%, or 80%,, or is a range defined by any two of the preceding values.
The microbial growth media of any one of the preceding claims, wherein the reduction in TTD of Candida is, is about, is at least, is at least about, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 75%, or 80%, or is a range defined by any two of the preceding values.
The microbial growth media of any one of the preceding claims, wherein the reduction in TTD of Candida is, is about, is at least, is at least about, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,12, 15, 18, 24, 30, 36, 48, 60, or 72 hours, or is a range defined by any two of the preceding values.
5 g/L TSB;
- 0.5 g/L SPS, 0.00005 g/L menadione, 0.01 g/L Pyridoxal HCl;
- 0.005 g/L ferulic acid;
- 0.7655 g/L sodium hydroxide;
- 0.05 g/L ascorbic acid;
- 0.0005 g/L L-cysteine, and 0.005 g/L hemin.
A culture container for detecting growth of a microbe, the container comprising:
- the microbial growth media of any one of the preceding claims;
- and
- a sensor for monitoring a parameter of the microbial growth media indicative of microbial growth in the microbial growth media.
A method of culturing a microbe in a sample, the method comprising:
- inoculating the microbial growth culture medium of any one of the preceding claims with a sample, and culturing a microbe in the sample in the microbial growth culture medium.
Dependencies are inferred from the claim text.