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Feeley Gorman Broth (F.G. Broth)

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SKU:
M812
Used for cultivation of Legionella species.

Intended Use

Recommended for cultivation of Legionella species.

Composition

Ingredients g/L
AcicaseTM# 17.500
HM peptone B ## 3.000
Starch 1.500
L-Cysteine hydrochloride 0.400
Ferric pyrophosphate, soluble 0.250
Final pH (at 25°C) 6.9±0.2

**Formula adjusted, standardized to suit performance parameters

#Equivalent to casein acid hydrolysate

## Equivalent to Beef extract

Directions

Suspend 22.65 grams in 1000 ml purified/distilled water. Heat if necessary to dissolve the medium completely. Dispense into tubes or flasks as desired. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes.

Principle And Interpretation

Feeley et al formulated (1,2) this medium, which is used as nonselective enrichment medium for isolation of Legionella species. Legionella is a gram-negative bacterium, including species that cause legionellosis or Legionnaires' disease, most notably L. pneumophilia (3). Legionella species are the causative agent of the human Legionnaires' disease and the lesser form, Pontiac fever. Legionella transmission occurs via aerosols- inhalation of mist droplets containing the bacteria. Common sources include cooling towers, domestic hot-water systems, fountains, and similar disseminators that tap into a public water supply. Natural sources of Legionella include freshwater ponds and creeks. Person-to-person transmission of Legionella has not been demonstrated (4).

Legionella are nutritionally fastidious and require L-cysteine and iron salts for their growth, which are provided in the medium. Legionella species are highly pathogenic microorganisms. Certain safety precautions must be taken when handling Legionella cultures.

AcicaseTM, HM peptone B, L-cysteine hydrochloride and ferric pyrophosphate act as sources of nutrients. Incubation should be carried out in the presence of 2.5% carbon dioxide but if it exceeds the limit, Legionella growth is inhibited due to formation of acidic condition. Legionella species can be identified by their characteristic fluorescence in presence of UV light (5,6). Since Legionella disease is primarily a pulmonary infection, prevention and containment of aerosols is essential (7).

Type of specimen

Clinical samples - Urine; Water samples

Specimen Collection and Handling:

For clinical samples follow appropriate techniques for handling specimens as per established guidelines (8,9).

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

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

After handling with Legionella species sample decontaminate work surface with either 5% hypochlorite or 5% phenil (carbolic acid).

Warning and Precautions :

In Vitro diagnostic use. 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 clinical specimens. Safety guidelines may be referred in individual safety data sheets.

Limitation

  1. Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
  2. Incubation should be carried out in the presence of 2.5% carbon dioxide but if it exceeds the limit, Legionella growth is inhibited due to formation of acidic condition.
  3. Further biochemical and serological tests must be performed for confirmation.

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

Colour and Clarity of prepared medium
Yellow coloured, clear to slightly opalescent solution in tubes

Reaction
Reaction of 2.3% w/v aqueous solution at 25°C. pH: 6.9±0.2

pH
6.70-7.10

Cultural Response

Cultural characteristics observed in presence of 2.5% Carbon dioxide (CO2) after an incubation at 35-37°C for 4 days .

Organism Growth Fluorescence under 366 nm
Legionella bozemanni ATCC 33217 good-luxuriant blue-white
Legionella micdadei ATCC 33218 good-luxuriant none
Legionella pneumophila ATCC 33153 good - luxuriant bright yellow

Storage and Shelf Life

Store between 10-30°C in a tightly closed container and the prepared medium at 15-30°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.

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 (8,9).

More Information
Product Name Feeley Gorman Broth (F.G. Broth)
SKU M812
Product Type Regular
Physical Form Powder
Origin Animal
Packaging type HDPE
References 1. Feeley J. C. et al, 1978, J. Clin. Microbiol., 8(3): 320.2.Feeley J. C. et al, 1979, J. Clin. Microbiol., 10(4):437.3.Ryan K. J., Ray C. G. (Eds.), 2004, Sherris Medical Microbiology, 4th Edition, McGraw Hill.4.Winn, W. C. Jr. ,1996, Legionella (In: Baron's Medical Microbiology, Barron, S. et al, (Eds.), 4th Edition, University ofTexas Medical Branch5.Herbert G. A. et al, 1959, Ann. Intern, Med., 92(1):45.6.Herbert G. A. et al, 1980, Ann. Intern. Med., 92(1):53.7.MacFaddin J. F., Vol. I, 1985, Media for Isolation Cultivation-Identification-Maintenance of Medical Bacteria, Williamsand Wilkins, Baltimore/ London,.
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