GN HiVeg™ Broth

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SKU:
MV242
For selective enrichment of gram-negative enteric organisms.


Intended use

Recommended for selective cultivation of gram-negative organisms of the enteric group.

Composition

Ingredients Gms / Litre
HiVeg™ hydrolysate No. 1 20.000
Dextrose (Glucose) 1.000
Mannitol 2.000
Sodium citrate 5.000
Synthetic detergent No. III 0.500
Dipotassium hydrogen phosphate 4.000
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 39.0 grams in 1000 ml distilled water. Heat if necessary to dissolve the medium completely. Dispense in test tubes or flasks as desired. Sterilize by autoclaving at A 115°C for 15 minutes. AVOID EXCESSIVE HEATING. A Corresponds to 10 lbs pressure.

Principle And Interpretation

Hajna (1 developed Gram Negative (GN Broth as an enrichment medium for recovery of Salmonella and Shigella from clinical and non-clinical specimens such as urine, blood clots, throat swabs, swabs from eating and drinking utensils etc (1,2. GN Broth, Hajna is also recommended by APHA (3 for the microbiological examination of foods. Croft and Miller isolated more strains of Shigella from rectal swabs using this medium (4. Taylor and Schelhart showed the superiority of GN Broth to selenite enrichment media for isolation of Shigella (5. Hajna (2,6 also suggested the enrichment of organisms from rectal swabs in this medium 1-6 hours before plating on solid media. GN HiVeg™ Broth is prepared by using vegetable peptones in place of animal based peptones which make the media free of BSE/TSE risks. The medium contains HiVeg™ hydrolysate No. 1, 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. The higher concentration of mannitol over dextrose limits the growth of Proteus and enhances growth of mannitol fermenting Salmonella and Shigella. This enrichment broth should be used in conjunction with selective and nonselective plating media to increase the probability of isolating pathogens (7,8,9. GN HiVeg™ Broth should be inoculated directly with the specimen. In case of stool specimens, approximately 1 gram should be used for inoculation. Appropriate references for processing of clinical and food samples should be followed (7,8,9,10. After incubation of 6-8 hours and again after 24 hours, sub culturing on selective agar media should be carried out (7.

Type of specimen

Please add specimens

Specimen Collection and Handling:

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 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: 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 an incubation at 35 - 37°C for 18 - 24 hours.

Organism Inoculum (CFU) Growth in GN broth Growth after 24 hours on MacConkey Agar Colour of colony
Escherichia coli ATCC 25922 (00013*) 50-100 good good pink-red with bile ppt
Enterococcus faecalis ATCC 19433 (00009*) 50-100 none-poor none-poor pale pink-red
Proteus mirabilis ATCC 25933 50-100 good good colourless
Pseudomonas aeruginosa ATCC 27853 (00025*) 50-100 good good colourless
Salmonella Typhimurium ATCC 14028 (00031*) 50-100 good good colourless
Shigella flexneri ATCC 12022 (00126*) 50-100 good good colourless

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. 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 sample must be decontaminated and disposed of in accordance with current laboratory techniques (11,12).

More Information
Product Name GN HiVeg™ Broth
SKU MV242
Product Type HiVeg™
Physical Form Powder
Origin Animal Free (Veg)
Packaging type HDPE
References 1. Hajna A. A., 1955, Publ. Health Lab., 13:59.
2.Hajna A. A., 1955, Publ. Health Lab., 13:83.
3.Salfinger Y., and Tortorello M.L., 2015, Compendium of Methods for the Microbiological Examination of Foods, 5thEd., American Public Health Association, Washington, D.C.
4.Hajna A. A., 1956, Air. Univ. Sch. Ar. Med., USAF, 56:39
5.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.
6.Forbes B. A., Sahm A. S., and Weissfeld D. F., Bailey & Scotts Diagnostic Microbiology, 10th Ed., 1998, Mosby, Inc.,St. Louis, Mo.
7.MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. 1, Williamsand Wilkins, Baltimore.
8.Ewing, 1986, Edwards and Ewings Identification of Enterobacteriaceae, 4th Ed., Elsevier Science Publishing Co., Inc., NewYork, N.Y.
9.Croft C. C., Miller M. J., 1956, Am. J. Clin. Pathol., 26:411.10.Taylor W.I., Schelhart D., 1968, Appl. Environ. Microbiol., 16:1383.
11. Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.1
2.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
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