CryoSaver™, Brucella Broth with Glycerol

Cat. no. CS81BNB CryoSaver™, Brucella Broth with Glycerol,
2ml Cryogenic Vial, 1.5ml Fill, Blue Cap
81 vials/box
Cat. no. CS100BNB CryoSaver™, Brucella Broth with Glycerol,
2ml Cryogenic Vial, 1.5ml Fill, Blue Cap
100 vials/box
Cat. no. CS81B CryoSaver™, Brucella Broth with Glycerol and Beads,
2ml Cryogenic Vial, 1.0 - 1.4ml Fill, Blue Cap
81 vials/box
Cat. no. CS100B CryoSaver™, Brucella Broth with Glycerol and Beads,
2ml Cryogenic Vial, 1.0 - 1.4ml Fill, Blue Cap
100 vials/box
Cat. no. CS100G CryoSaver™, Brucella Broth with Glycerol and Beads,
2ml Cryogenic Vial, 1.0 - 1.4ml Fill, Green Cap
100 vials/box
Cat. no. CS100N CryoSaver™, Brucella Broth with Glycerol and Beads,
2ml Cryogenic Vial, 1.0 - 1.4ml Fill, Natural Cap
100 vials/box
Cat. no. CS100R CryoSaver™, Brucella Broth with Glycerol and Beads,
2ml Cryogenic Vial, 1.0 - 1.4ml Fill, Red Cap
100 vials/box
Cat. no. CS100Y CryoSaver™, Brucella Broth with Glycerol and Beads,
2ml Cryogenic Vial, 1.0 - 1.4ml Fill, Yellow Cap
100 vials/box

INTENDED USE

Hardy Diagnostics CryoSaver™, Brucella Broth with Glycerol is recommended for the preservation of microorganisms by freezing.

This product is not intended to be used for the diagnosis of human disease.

SUMMARY

CryoSaver™, Brucella Broth with Glycerol is formulated using 10% glycerol. The high concentration of glycerol acts to prevent cell damage in the harsh environment of sub-zero temperatures. CryoSaver™, Brucella Broth with Glycerol and Beads, contains approximately 20 beads. The cleaned, acid-washed beads are of porous nature allowing microorganisms to readily adhere to the bead surface. When a fresh culture is required, a single bead is easily removed from the vial and used to directly inoculate a suitable bacteriological medium. Alternatively, a spatula may be used to scrape the frozen broth for use as an inoculum. Stock cultures stored in the CryoSaver™, Brucella Broth with Glycerol can be kept for one year if stored at -50°C. Stock cultures maintained at -70°C, or in liquid nitrogen, may be kept indefinitely.(1)

The CryoSaver™ system is designed for ease of use. Numbered compartments in a durable container protect the CryoSaver™ vials and saves space in the freezer. Each vial has an easy "write-on" label for specimen identification. Accesories include the workstation, which securely holds up to 40 vials for easy one-handed operation when screwing the caps on and off. Inoculation needles are available to aid in the removal of the beads from the CryoSaver™ vial. A sharp edged spatula is available for the microbiologist that prefers to scrape the frozen culture rather than to use the beads. Colored round cap inserts are available for color coding of the vials. These inserts are available in eleven colors. In the CS100 product line, the caps themselves are available in five colors. See the "Materials Required But Not Provided" section for more information.

Brucella Broth is a highly nutritious general purpose growth medium for fastidious and non-fastidious organisms. It contains pancreatic digest of casein, peptic digest of animal tissue, and yeast extract as a rich source of nutrients.

FORMULA

Ingredients per 900ml of deionized water:*

Pancreatic Digest of Casein 10.0gm
Peptic Digest of Animal Tissue 10.0gm
Sodium Chloride 5.0gm
Yeast Extract 2.0gm
Dextrose 1.0gm
Sodium Bisulfite 0.1gm
Glycerol 100.0ml

Final pH 7.0 +/- 0.3 at 25°C.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-30°C away from direct light. Media should not be used if there are any signs of contamination, deterioration, discoloration, or if the expiration date has passed. Product is light and temperature sensitive.

PRECAUTIONS

PROCEDURE - FOR USE OF BEADS

Preparation

1. Prepare an overnight culture of the organism to be stored on an appropriate non-selective medium, such as TSA (Cat. no. G60) or Blood Agar (Cat. no. A10). Do not use cultures older than 24 hours.

2. Label the vial with the date, name, and source of the organism. Also write in the name on the card, included in the CryoSaver™ box.

3. Using a sterile loop and appropriate aseptic technique, transfer several colonies into the vial, resulting in a dense suspension.

4. Shake or vortex well. To ensure that the cryoprotectant has penetrated the bacterial cells, allow the suspension to sit at room temperature for a minimum of 15 minutes, but no longer than 45-60 minutes.(6)

5. If you prefer to use the bead method, draw out all of the liquid portion from the vial using a disposable sterile pipet (Cat. no. 2221S). Discard the pipet and its liquid contents into an appropriate biohazard container for sterilization. Close the vial finger tight. If you prefer the scraping method see "Alternate Procedure" below.

6. Store the vial with beads at -50 or -70°C.
Cultures stored at -50°C can be kept for one year.
Cultures stored at -70°C, or in liquid nitrogen may be kept indefinitely.

Recovery

7. Under aseptic conditions, open the vial and remove one of the colored beads using a sterile nichrome needle (Cat. no. 3060), a disposable plastic needle (Cat. no. HSND), or forceps. Quickly re-cap the vial and return it immediately to the freezer. This task must be performed quickly. Do not allow the contents of the vial to thaw.

8. This inoculum may then be used to directly streak onto a solid non-selective medium (e.g. Chocolate Agar, Cat. no. E14) or may be dropped into an appropriate liquid medium. Incubate the culture media at an appropriate temperature and atmosphere for the organism to be recovered.

ALTERNATE PROCEDURE - FOR SCRAPING WITH A SPATULA

This procedure is for those who prefer to scrape the top of the frozen vial with a spatula, rather than using the beads. For the inoculation of the culture media.

Preparation

Follow the above steps 1 through 4. Omit step 5 and continue with step 6.

Recovery

Omit step 7. Open the vial and transfer the organism by scraping off some frozen shavings onto the plated culture media. For best results use a stainless steel spatula with a sharp edge (Cat. no. 57949041). Quickly re-cap the vial and return it immediately to the freezer. Do not allow the contents of the vial to thaw. Proceed to step 8 above.

See listed references for more information on storage of stock cultures. (1-4)

LIMITATIONS

CryoSaver™, Brucella Broth with Glycerol is a medium for the long-term storage of bacteria. It is not suitable for use in the identification of microorganisms.

Although cultures may be kept indefinitely at -70°C, it is recommended to replace non-fastidious organisms every 5 years and fastidious organisms every 3 years. (1)

Aseptic technique should be practiced to ensure continued integrity and purity of the stored microorganism.

This media should not be used if any of the following conditions are present before inoculation:
- the vial shows any evidence of leakage (loss of broth)
- the vial shows any evidence of turbidity, which suggests contamination
- the expiration date has elapsed

After removal, the beads should not be returned to the vial for any reason.

Do not thaw and refreeze the vial.

MATERIALS REQUIRED BUT NOT PROVIDED

The following accessories are available for use with this system:

802501 CryoSaver™ Workstation, securely holds 40 vials 1 workstation
57949041 Spatula, stainless steel, 16.8cm long 1 spatula
3060 Needle, nichrome with aluminum handle and insulated steel 5 needles/pack
HSND Needle, disposable, plastic, rigid, violet color 25 needles/pack
T3121 CryoSaver™ Cap Insert, White 500 caps/bag
T3122 CryoSaver™ Cap Insert, Blue 500 caps/bag
T3123 CryoSaver™ Cap Insert, Red 500 caps/bag
T3124 CryoSaver™ Cap Insert, Green 500 caps/bag
T3125 CryoSaver™ Cap Insert, Yellow 500 caps/bag
T3128 CryoSaver™ Cap Insert, Tan 500 caps/bag
T3129 CryoSaver™ Cap Insert, Gray 500 caps/bag
T31210 CryoSaver™ Cap Insert, Lilac 500 caps/bag
T31211 CryoSaver™ Cap Insert, Orange 500 caps/bag
T31213 CryoSaver™ Cap Insert, Violet 500 caps/bag
T31214 CryoSaver™ Cap Insert, Pink 500 caps/bag
T3127 CryoSaver™ Cap Insert, Assorted 1,000 caps/bag

Standard microbiological supplies and equipment such as freezers, loops, pipets, needles, other culture media, swabs, applicator sticks, incinerators, and incubators, etc., are not provided.

QUALITY CONTROL

Test Organisms Results
Escherichia coli
ATCC® 25922
Growth upon subculture to TSA after overnight freezing at -70°C.
Staphylococcus epidermidis
ATCC® 12228
Growth upon subculture to TSA after overnight freezing at -70°C.
Streptococcus pyogenes
ATCC® 19615
Growth upon subculture to Blood Agar, 5% after overnight freezing at -70°C.

User Quality Control

PHYSICAL APPEARANCE

CryoSaver™, Brucella Broth

CryoSaver™, Brucella Broth with Glycerol and Beads, 2ml Cryogenic Vial, 1.0 - 1.4ml Fill, Blue Cap (Cat. no. CS100B).

CryoSaver™, Brucella Broth

CryoSaver™, Brucella Broth with Glycerol and Beads, 2ml Cryogenic Vial, 1.0 - 1.4ml Fill, Red Cap (Cat. no. CS100R).

CryoSaver™, Brucella Broth

CryoSaver™, Brucella Broth with Glycerol Vial with Beads.

CryoSaver™, Brucella Broth

Showing inoculation with wire needle (Cat. no. 3060) and bead from CryoSaver™, Brucella Broth with Glycerol and Beads.

REFERENCES

1. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.

2. Versalovic, J., et al. Manual of Clinical Microbiology. American Society for Microbiology, Washington, D.C.

3. Tille, P., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

4. Koneman, E.W., et al. Color Atlas and Textbook of Diagnostic Microbiology, J.B. Lippincott Company, Philadelphia, PA.

5. Quality Assurance for Commercially Prepared Microbiological Culture Media, M22. Clinical and Laboratory Standards Institute (CLSI - formerly NCCLS), Wayne, PA.

6. ATCC connection, Vol. 26, No. 1. 2006. p. 4-5.


ATCC is a registered trademark of the American Type Culture Collection.

051216gr