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Blood HiCynth™ Agar Base (Infusion HiCynth™ Agar)

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MCD073
Recommended for isolation and cultivation of many fastidious pathogenic microorganisms like Neisseria, Streptococci after addition of blood from clinical and non-clinical specimens.

Intended Use:

Recommended for isolation and cultivation of many fastidious pathogenic microorganisms after addition of blood.

Composition**

Ingredients Gms / Litre
HiCynth™ Peptone No.3* 20.000
Sodium chloride 5.000
Agar 15.000
Final pH (at 25°C) 7.3±0.2

**Formula adjusted, standardized to suit performance parameters

*Chemically defined peptone

Directions

Suspend 40.0 grams in 1000 ml purified/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 HiCynth™ Agar Base is a highly nutritious medium generally used as a basal medium for preparing blood agar by supplementation with blood. It is prepared by completely replacing animal or vegetable peptones with chemically defined peptones to avoid BSE/TSE risks associated with animal peptones. 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.

HiCynth™ Peptone No.3 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 hemolysis and mimic Streptococcus pyogenes (9).

Type of specimen

Food samples; Water samples

Specimen Collection and Handling

For food samples, follow appropriate techniques for sample collection and processing as per guidelines (5).

For water samples, follow appropriate techniques for sample collection and processing as per guidelines (10).

After use, contaminated materials must be sterilized by autoclaving before discarding.

Warning and Precautions

For professional 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 specimens. Safety guidelines may be referred in individual safety data sheets.

Limitations

  1. Addition of sheep blood is recommended to detect hemolysis. 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 β- hemolytic Streptococci and hence must be confirmed.
  3. Haemolytic pattern varies with the source of blood used.
  4. Further biochemical and serological testing is required for complete identification.

Performance and Evaluation

Performance 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% v/v sterile defibrinated blood, after an incubation at 35-37°C for 18-48 hours.

Organism Inoculum (CFU) Growth w/o blood Recovery w/o blood Growth with blood Recovery with blood Haemolysis
Neisseria meningitidis ATCC 13090 50-100 fair 40-50% luxuriant >=70% none
Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) 50-100 good 50-70% luxuriant >=70% beta
Staphylococcus epidermidis ATCC 12228 (00036*) 50-100 good 50-70% luxuriant >=70% none
Streptococcus pneumoniae ATCC 6303 50-100 fair-good 40-50% luxuriant >=70% alpha
Streptococcus pyogenes ATCC 19615 50-100 fair-good 40-50% luxuriant >=70% beta

Key: (*) Corresponding WDCM numbers.

Storage and Shelf Life

Store between 10-30°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. 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 sample must be decontaminated and disposed of in accordance with current laboratory techniques (11,12).

More Information
Product Name Blood HiCynth™ Agar Base (Infusion HiCynth™ Agar)
SKU MCD073
Product Type HiCynth™
Physical Form Powder
Origin Chemically defined (HiCynth™)
Packaging type HDPE
References 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 ofFoods, 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 ClinicalMicrobiology, ASM, Washington, D.C.10.Isenberg, H.D. Clinical Microbiology Procedures Handb0ook. 2nd Editio11. 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.
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