Please refer disclaimer Overleaf.

M431

Composition**Ingredients Gms / LitreTryptone 17.000Soya peptone 3.000Sodium chloride 5.000Dextrose (Glucose) 2.500Dipotassium hydrogen phosphate 2.500Sodium azide 0.500Agar 10.000Final pH ( at 25°C) 7.4±0.2**Formula adjusted, standardized to suit performance parameters # Equivalent to Papaic digest of soyabean meal

DirectionsSuspend 40.5 grams in 1000 ml purified/distilled water. If desired add 3 more grams of agar. Heat with frequent agitation to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. Mix well and pour into sterile Petri plates or dispense as desired.

Principle And InterpretationAnaerobic bacteria are widely distributed in nature in oxygen-free habitats. When collecting specimens from patients for isolation and identification of anaerobic bacteria associated with infections, care should be taken that the specimens should be free of contaminating bacteria. Material from sites that are normally sterile, such as blood, spinal fluid, or pleural fluid, poses no problem provided the usual precautions are taken to decontaminate the skin properly before puncturing it to obtain the specimen. Fecal specimens, sputum specimens, or vaginal secretions cannot be cultured routinely for pathogenic anaerobes because they normally contain other aerobic organisms. Aspirates from abscesses or the specific sites of infections must be obtained in these cases to avoid undue contamination with indigenous flora components. Forget-Fredette Agar is based on the formulation described by Fredette et al (1) and Forget and Fredette (2), and it is used for selective isolation of anaerobic microorganisms from a mixture of aerobic and anaerobic flora. Forget and Fredette employed this medium to study the anaerobic flora in chronic nasal sinusitis.Soya peptone and tryptone provide the necessary nutrients to the anaerobic microorganisms. Dextrose is the carbohydrate source. Dipotassium phosphate buffers the medium. Sodium azide inhibits not only gram-negative cocci and bacilli but also most of the gram-positive aerobes. Forget Fredette Agar allows the growth of Clostridia, Fusiforms, Ristella, Sphaerophorus and Streptococci in the depths of the medium tubes. Bacillus species, Listeria and most gram-negative organisms are inhibited on this medium. This medium can also be used for surface cultivation of anaerobes in plates in anaerobic jars. However for this purpose, it is recommended to add about four grams of agar to the medium before sterilization. The test sample is inoculated into the depths of the medium tube to facilitate isolation of anaerobic bacteria.

Forget Fredette AgarIntended Use:Recommended for selective isolation of anaerobes from a mixture of aerobic and anaerobic flora.

Type of specimen Decaying vegetables, soil, cheese

For food samples follow appropriate techniques for handling specimens as per established guidelines (5). After use, contaminated materials must be sterilized by autoclaving before discarding.

Specimen Collection and Handling

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. Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.

HiMedia Laboratories Technical Data

Quality ControlAppearanceCream to yellow homogeneous free flowing powderGellingFirm,comparable with 1.0% Agar gel.Colour and Clarity of prepared mediumYellow coloured, clear to slightly opalescent gel forms in tubesReactionReaction of 4.05% w/v aqueous solution at 25°C. pH : 7.4±0.2pH7.20-7.60Cultural ResponseCultural characteristics observed after an incubation at 35-37°C for 24-48 hours (Clostridium species incubated anaerobically).

Organism Inoculum(CFU)

Growth

Clostridium perfringens ATCC 12924

50-100 good-luxuriant

Streptococcus pneumoniae ATCC 6303

50-100 good-luxuriant

Reference1. Fredette, Auger and Forget, 1962, J. Bacterial, June 83 (6), 1217-1223 Sodium oxide selective medium for the primary

isolation of Anaerobic bacteria.

2. Forget and Fredette, 1962, J. Bacteriol., 83:121.3. 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) Manualof Clinical Microbiology, 11th Edition. Vol. 1.

Revision :02/2021

Disclaimer :

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

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

Storage and Shelf LifeStore 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. Product performance is best if used within stated expiry period.

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 (3,4).

Disposal

HiMedia Laboratories Pvt. Ltd. Reg.office : 23, Vadhani Ind.Est., LBS Marg, Mumbai-400086, India. Customer care No.: 022-6116 9797 Corporate office : 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 Website: www.himedialabs.com

2. Each lot of the medium has been tested for the organisms specified on the COA. It is recommended to users to validate the medium for any specific microorganism other than mentioned in the COA based on the user’s unique requirement.

5. Salfinger Y., and Tortorello M.L., 2015, Compendium of Methods for the Microbiological Examination of Foods, 5thEd., American Public Health Association, Washington, D.C.