Blood Agar Base (Infusion Agar) M073

Intended Use:
Blood Agar Base is recommended as a base to which blood may be added for use in the isolation and cultivation of
fastidious pathogenic microorganisms like Neisseria , Streptococci etc.
Composition**
Ingredients Gms / Litre
HM peptone B# 10.000
Tryptose 10.000
Sodium chloride 5.000
Agar 15.000
Final pH ( at 25°C) 7.3±0.2
**Formula adjusted, standardized to suit performance parameters
# Equivalent to Beef Heart peptone
Directions
Suspend 40.0 grams in 1000 ml distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving
at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C and aseptically add 5% v/v sterile defibrinated blood. Mix well
and pour into sterile Petri plates.

Principle And Interpretation

Blood Agar Base is a highly nutritious medium generally used as a basal medium for preparing blood agar by supplementation
with blood. It can also be used as general-purpose media without the addition of blood.
Blood Agar Base media can be used with added phenolphthalein phosphate (1) for the detection of phosphate producing
Staphylococci, with added salt and agar for assessment of surface contamination on equipment and pig carcass (2) and to
determine salinity range of marine Flavobacteria (3). It can also be used for preparation of Salmonella Typhi antigens (4).
Blood Agar Base is recommended by APHA (5) and Standard Methods (6, 7) for testing of food samples.
HM peptone B and tryptose provides carbon, nitrogen, amino acids and vitamins. Sodium chloride helps in maintaining the
osmotic equilibrium of the medium. Addition of blood makes the medium more nutritious by providing additional growth
factors required by fastidious organisms. It also helps in visualizing the haemolytic reactions. However, haemolytic reactions
depend on the animal blood used. Sheep blood gives best results for Group A Streptococci (8). But sheep blood fails to
support growth of Haemophilus haemolyticus since sheep blood is deficient in pyridine nucleotides. However when horse
blood is used H. haemolyticus colonies produce haemolysis and mimic Streptococcus pyogenes (9).

Type of specimen
Clinical material : blood and other pathological material ; food samples

Specimen Collection and Handling

For clinical samples follow appropriate techniques for handling specimens as per established guidelines (9,10,11).
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (5,6).
After use, contaminated materials must be sterilized by autoclaving before discarding.

Warning and Precautions

In Vitro diagnostic use only. Read the label before opening the container. Wear protective gloves/protective
clothing/eye protection/face protection. Follow good microbiological lab practices while handling specimens and culture.
Standard precautions as per established guidelines should be followed while handling clinical specimens. Safety guidelines
may be referred in individual safety data sheets.

Please refer disclaimer Overleaf.

HiMedia Laboratories Technical Data

Limitations
1.Addition of sheep blood is recommended to detect haemolysis. This medium does not support the growth of H.haemolyticus
2.Addition of Horse blood or rabbit blood to base medium supports growth of H.haemolyticus but resemble beta-
haemolytic Streptococci and hence must be confirmed.
3. Haemolytic pattern varies with the source of blood used.

Performance and Evaluation

Performace of the medium is expected when used as per the direction on the label within the expiry period when stored at
recommended temperature.

Quality Control
Appearance
Cream to yellow homogeneous free flowing powder
Gelling
Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium
Basal medium : Light amber coloured clear to slightly opalescent gel. After addition of 5% v/v sterile defibrinated blood :
Cherry red coloured opaque gel forms in Petri plates.
Reaction
Reaction of 4.0% w/v aqueous solution at 25°C. pH : 7.3±0.2
pH
7.10-7.50
Cultural Response
Cultural characteristics observed with added 5% w/v sterile defibrinated blood,after an incubation at 35-37°C for 18-48
hours.

Organism Inoculum Growth w/o Recovery w/o Growth with Recovery with Haemolysis
(CFU) blood blood blood blood
Cultural Response
Neisseria meningitidis ATCC 50-100 fair 40-50% luxuriant >=70% none
13090
Staphylococcus aureus 50-100 good 50-70% luxuriant >=70% beta
subsp. aureus ATCC
25923 (00034*)

Staphylococcus epidermidis 50-100 good 50-70% luxuriant >=70% none

ATCC 12228 (00036*)
Streptococcus pneumoniae 50-100 fair-good 40-50% luxuriant >=70% alpha
ATCC 6303
Streptococcus pyogenes 50-100 fair-good 40-50% luxuriant >=70% beta
ATCC 19615
Key : *Corresponding WDCM numbers.

Storage and Shelf Life

Store below 30°C in a tightly closed container and the prepared medium at 2 - 8°C. Use before expiry date on the label. On
opening, product should be properly stored dry, after tightly capping the bottle in order to prevent lump formation due to the
hygroscopic nature of the product. Improper storage of the product may lead to lump formation. Store in dry ventilated area
protected from extremes of temperature and sources of ignition Seal the container tightly after use. Use before expiry date on
the label. Product performance is best if used within stated expiry period.

Disposal
User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow
established laboratory procedures in disposing of infectious materials and material that comes into contact with clinical
sample must be decontaminated and disposed of in accordance with current laboratory techniques (10,11).

Please refer disclaimer Overleaf.

HiMedia Laboratories Technical Data

Reference
1. Noble W. C., 1962, J. Clin, Pathol., 15:552.
2. Hansen N. H., 1962, J. Appl. Bacteriol., 25:46.
3. Hayes P. R., 1963, J. Gen. Microbiol., 30:1.
4. Schuber J. H., Edwards P. R. and Ramsere C. H., 1959, J. Bacteriol., 77:648.
5. Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2015, Compendium of Methods for the Microbiological Examination of
Foods, 5th Ed., American Public Health Association, Washington, D.C.
6.U.S. Food and Drug Administration, 1995, Bacteriological Analytical Manual, 8th Ed., AOAC International, Gaithersburg,
Md.
7.Atlas R. M., 1993, Handbook of Microbiology of Microbiological Media, CRC Press, Boca Raton, Fla.
8.Snavely J. G. and Brahier J., 1960, Am. J. Clin. Pathol., 33:511.
9.Murray P. R., Baron J. H., Jorgensen J. H., Pfaller M. A., Yolken R. H., (Eds.), 8th Ed., 2003, Manual of Clinical
Microbiology, ASM, Washington, D.C.
10.Isenberg, H.D. Clinical Microbiology Procedures Handb0ook. 2nd Editio
11. Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015)
Manual Clinical Microbiology, 11th Edition. Vol. 1.

Revision : 03 / 2018

In vitro diagnostic medical

IVD device

CE Marking

30°C Storage temperature

10°C

Do not use if package is

damaged

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LBS Marg,Mumbai-86,MS,India

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Disclaimer :
User must ensure suitability of the product(s) in their application prior to use. Products conform solely to the information contained in
this and other related HiMedia™ publications. The information contained in this publication is based on our research and development
work and is to the best of our knowledge true and accurate. HiMedia™ Laboratories Pvt Ltd reserves the right to make changes to
specifications and information related to the products at any time. Products are not intended for human or animal or therapeutic use but
for laboratory,diagnostic, research or further manufacturing use only, unless otherwise specified. Statements contained herein should not
be considered as a warranty of any kind, expressed or implied, and no liability is accepted for infringement of any patents.

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