Nutrient Agar M001
Intended use
Nutrient Agar is used as a general purpose medium for the cultivation of less fastidious microorganisms, can be enriched
with blood or other biological fluids.
Composition**
Ingredients Gms / Litre
Peptone 5.000
Sodium chloride 5.000
HM peptone B# 1.500
Yeast extract 1.500
Agar 15.000
Final pH ( at 25°C) 7.4±0.2
**Formula adjusted, standardized to suit performance parameters
# - Equivalent to Beef extract
Directions
Suspend 28.0 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. If desired ,the medium can be enriched with
5-10% blood or other biological fluids. Mix well and pour into sterile Petri plates.
Principle And Interpretation
Nutrient media are basic culture media used for maintaining microorganisms, cultivating fastidious organisms by enriching
with serum or blood and are also used for purity checking prior to biochemical or serological testing (5,6). Nutrient Agar is
ideal for demonstration and teaching purposes where a more prolonged survival of cultures at ambient temperature is often
required without risk of overgrowth that can occur with more nutritious substrate. This relatively simple formula has been
retained and is still widely used in the microbiological examination of variety of materials and is also recommended by
standard methods. It is one of the several non-selective media useful in routine cultivation of microorganisms (1,7). It can be
used for the cultivation and enumeration of bacteria which are not particularly fastidious. Addition of different biological
fluids such as horse or sheep blood, serum, egg yolk etc. makes it suitable for the cultivation of related fastidious organisms.
Peptone, HM peptone B and yeast extract provide the necessary nitrogen compounds, carbon, vitamins and also some trace
ingredients necessary for the growth of bacteria. Sodium chloride maintains the osmotic equilibrium of the medium.
Type of specimen
Clinical samples - Blood; Food and dairy samples; Water samples
Specimen Collection and Handling:
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (3,4).
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (1,7,8).
For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards.(2)
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.
Limitations :
1.This medium is general purpose medium and may not support the growth of fastidious
organisms.
Please refer disclaimer Overleaf.
HiMedia Laboratories Technical Data
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
Light yellow coloured clear to slightly opalescent gel forms in Petri plates
Reaction
Reaction of 2.8% w/v aqueous solution at 25°C. pH : 7.4±0.2
pH
7.20-7.60
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-48 hours.
Organism Inoculum Growth Recovery
(CFU)
Escherichia coli ATCC 50-100 good-luxuriant >=70%
25922 (00013*)
Pseudomonas aeruginosa 50-100 good-luxuriant >=70%
ATCC 27853 (00025*)
Salmonella Typhi ATCC 50-100 good-luxuriant >=70%
6539
Staphylococcus aureus 50-100 good-luxuriant >=70%
subsp.aureus ATCC
25923 (00034*)
Streptococcus pyogenes 50-100
ATCC 19615 good-luxuriant >=70%
Salmonella Enteritidis 50-100 good-luxuriant >=70%
ATCC 13076 (00030*)
Salmonella Typhimurium 50-100 good-luxuriant >=70%
ATCC 14028 (00031*)
Yersinia enterocolitica 50-100 good-luxuriant >=70%
ATCC 9610 (00038*)
Yersinia enterocolitica 50-100 good-luxuriant >=70%
ATCC 23715 (00160*)
Key : *Corresponding WDCM numbers.
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 20-30°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 (4,5).
Please refer disclaimer Overleaf.
HiMedia Laboratories Technical Data
Reference
1. American Public Health Association, Standard Methods for the Examination of Dairy Products, 1978, 14th Ed.,
Washington D.C.
2. Baird R.B., Eaton A.D., and Rice E.W., (Eds.), 2015, Standard Methods for the Examination of Water and Wastewater,
23rd ed., APHA, Washington, D.C.
3. Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
4. 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.
5. Lapage S., Shelton J. and Mitchell T., 1970, Methods in Microbiology', Norris J. and Ribbons D., (Eds.), Vol. 3A,
Academic Press, London.
6. MacFaddin J. F., 2000, Biochemical Tests for Identification of Medical Bacteria, 3rd Ed., Lippincott, Williams and
Wilkins,Baltimore.
7. 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.
8. Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th
Ed.,APHA Inc., Washington, D.C.
Revision : 05 / 2019
In vitro diagnostic medical
IVD device
CE Marking
30°C Storage temperature
10°C
Do not use if package is
damaged
HiMedia Laboratories Pvt. Limited,
23 Vadhani Industrial Estate,
LBS Marg,Mumbai-86,MS,India
CE Partner 4U ,Esdoornlaan 13, 3951
EC REP
DB Maarn The Netherlands,
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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.
HiMedia Laboratories Pvt. Ltd. Reg.office : 23, Vadhani Ind.Est., LBS Marg, Mumbai-400086, India. Customer care No.: 022-6116 9797 Corporate
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