GN Broth Medium 11.

MM242
Intended Use:
For enrichment of Shigella from pharmaceutical products in accordance with Indian pharmacopoeia 2017.
Composition**
Ingredients Gms / Litre
Polypeptone peptone 20.000
Glucose 1.000
Sodium citrate 2.000
Sodium deoxycholate 0.500
Di-potassium hydrogen phosphate 4.000
Mono potassium dihydrogen phosphate 1.500
Sodium chloride 5.000
Final pH ( at 25°C) 7.0±0.2
**Formula adjusted, standardized to suit performance parameters

Directions
Suspend 34.00 grams in 1000 ml purified/ distilled water. Mix well and allow to stand for 15 minutes. With continuous stirring
bring gently to boil and maintain at boiling till completely dissolved. DO NOT AUTOCLAVE. Dispense in sterile test tubes
or flasks as desired.

Principle And Interpretation

GN Broth is recommended by the Indian Pharmacopoeia (1) for the selective isolation of Shigella species with subsequent
isolation on a selective medium, XLD Agar (MH031) .Croft and Miller isolated more strains of Shigella from rectal swabs
using this medium (2). Taylor and Schelhart showed the superiority of GN Broth to selenite enrichment media for isolation of
Shigella (3). Hajna (4,5) also suggested the enrichment of organisms from rectal swabs in this medium 1-6 hours before
plating on solid media.
The medium contains polypeptone peptone, which provides amino acids and other nitrogenous substances to support
bacterial growth. The combination of sodium citrate and sodium deoxycholate inhibit gram-positive and some gram-negative
bacteria such as coliforms. Phosphates serve as a buffering system.Sodium chloride maintains osmotic equilibrium. Proteus,
Pseudomonas and coliforms do not overgrow Salmonella and Shigella in GN Broth during the first 6 hours of incubation.
This enrichment broth should be used in conjunction with selective and nonselective plating media to increase the probability
of isolating pathogens (6,7,8).

Type of specimen
Pharmaceutical samples

Specimen Collection and Handling

For pharmaceutical samples, follow appropriate techniques for sample collection, processing as per guidelines (1).
After use, contaminated materials must be sterilized by autoclaving before discarding.

Warning and Precautions

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. Further isolation and biochemical tests must be carried out for confirmation.

Please refer disclaimer Overleaf.

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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
Colour and Clarity of prepared medium
Light amber coloured, clear to slightly opalescent solution in tubes.
Reaction
Reaction of 3.9% w/v aqueous solution at 25°C. pH : 7.0±0.2
pH
6.80-7.20
Cultural Response
Cultural characteristics observed after inoculation in GN Broth and then subculture on XLD Agar and incubation at 30-35°C
for 24-48 hours.
Growth promoting properties
Growth promotion was carried out in accordance with the method of IP.Clearly visible growth of microorganism comparable
to that previously obtained with previously tested and approved lot of medium occurs at the specified temperature for not
more than the shortest period of time specified inoculating <=100 cfu (at 30-35°C for <=24 hours).
Inhibitory properties
No growth of the test microorganism occurs for the specified temperature for not less than longest period of time specified
inoculating >=100 cfu (at least 100 cfu) (at 30-35°C for >=48 hours).
Cultural Response
Organism Inoculum Growth in GN Recovery on Recovery Colour of
(CFU) broth XLD Agar colony
Growth promoting
Shigella boydii ATCC 8700 50 -100 good good- >=50 % red
Inhibitory luxuriant translucent
Staphylococcus aureus >=10³ inhibited <=0 %
ATCC 6538 (00193*)
* - 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 (6,7).

Reference
1.Indian Pharmacopoeia, 2017, Ministry of Health and Family Welfare, Govt. of India,.
2.Croft C. C., Miller M. J., 1956, Am. J. Clin. Pathol., 26:411.
3.Taylor W.I., Schelhart D., 1968, Appl. Environ. Microbiol., 16:1383.
4.Hajna A. A., 1955, Publ. Health Lab., 13:83.
5.Hajna A. A., 1956, Air. Univ. Sch. Ar. Med., USAF, 56:39.
6.Downes F. P. and Ito K., (Eds.), 2001, Compendium of Methods for the Microbiological Examination of Foods, 4th Ed.,
American Public Health Association, Washington, D.C.
7. Murray P. R., Baron J. H., Pfaller M. A., Jorgensen J. H. and Yolken R. H., (Ed.), 2003, Manual of Clinical Microbiology,
8th Ed., American Society for Microbiology, Washington, D.C.
8.MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. 1, Williams
and Wilkins, Baltimore.
9..Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition.
Please refer disclaimer Overleaf.
 HiMedia Laboratories Technical Data

10. Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015)
Manual of Clinical Microbiology, 11th Edition. Vol. 1

Revision : 03 / 2018

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
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