Reinforced Clostridial Broth M443
Reinforced Clostridial Broth is used for the cultivation and enumeration of Clostridia and other anaerobes.
Composition**
Ingredients Gms / Litre
Casein enzymic hydrolysate 10.000
Beef extract 10.000
Yeast extract 3.000
Dextrose 5.000
Sodium chloride 5.000
Starch, soluble 1.000
L-Cysteine hydrochloride 0.500
Sodium acetate 3.000
Agar 0.500
Final pH ( at 25°C) 6.8±0.2
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 38.00 grams in 1000 ml distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving
at 10 lbs pressure (115°C) for 15 minutes. Mix well and dispense as desired.
Principle And Interpretation
Reinforced Clostridial Broth is formulated by Hirsch and Grinsted (1). It can be used to initiate growth from small inocula and
to obtain the highest viable count of Clostridia . Barnes and Ingram used the broth medium for diluting an inoculum of
vegetative cells of Clostridium perfringens (2). It can be used in studies of spore forming anaerobes, especially Clostridium
butyricum in cheese, for enumeration of Clostridia in tube dilution counts or for preparation of plates for isolation (3). Other
spore forming anaerobes, Streptococci and Lactobacilli also grow in this media. This is a nonselective enrichment media.
Casein enzymic hydrolysate, yeast extract, beef extract, starch, L-cysteine and sodium acetate provide all the necessary nutrients
for the growth of Clostridia . Dextrose is a fermentable carbohydrate in the medium while sodium chloride maintains osmotic
equilibrium. This media can be made selective by addition of 15-20 mg Polymyxin B per litre of media (1).
Quality Control
Appearance
Cream to yellow homogeneous free flowing powder
Colour and Clarity of prepared medium
Light yellow coloured clear solution in tubes.
Reaction
Reaction of 3.8% w/v aqueous solution at 25°C . pH : 6.8±0.2
pH
6.60-7.00
Cultural Response
Cultural characteristics observed in an anaerobic atmosphere after an incubation at 35 - 37°C for 24 - 48hours.
Cultural Response
Organism Inoculum Growth
(CFU)
Cultural Response
Clostridium sporogenes 50 -100 good - luxuriant
ATCC 11437
Please refer disclaimer Overleaf.
HiMedia Laboratories Technical Data
Clostridium sporogenes 50 -100 good - luxuriant
ATCC 19404
Bacteroides vulgatus ATCC 50 -100 good - luxuriant
8482
Bacteroides fragilis ATCC 50 -100 good - luxuriant
23745
Clostridium perfringenes 50 -100 good - luxuriant
ATCC 13124
Storage and Shelf Life
Store below 30°C in tightly closed container and the prepared medium at 2 - 8°C. Use before expiry date on the label.
Reference
1.Hirsch and Grinsted, 1954, J. Dairy Res., 21:101.
2.Barnes and Ingram, 1956, J. Appl. Bact., 19:117.
3.Lewis and Angelotti (Eds.), 1964, Examination of Foods for Enteropathogenic and Indicator Bacteria, Dept. of HEW, PHS
Publication, 1142, Washington.
Revision : 2 / 2015
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. A-516,Swastik Disha Business Park,Via Vadhani Ind. Est., LBS Marg, Mumbai-400086, India. Customer care No.: 022-6147
1919 Email: techhelp@himedialabs.com
