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Hi-Sensitivity™ Test HiVeg™ Agar

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MV485

for antimicrobial susceptibility tests.

Description

Intended use

Recommended for antimicrobial susceptibility tests.

Composition**

Ingredients	g/L
HiVeg™ hydrolysate	11.000
HiVeg™ peptone	3.000
Dextrose (Glucose)	2.000
Sodium chloride	3.000
Starch, soluble	1.000
Disodium hydrogen phosphate	2.000
Sodium acetate	1.000
Magnesium glycerophosphate	0.200
Calcium gluconate	0.100
Cobaltous sulphate	0.001
Cupric sulphate	0.001
Zinc sulphate	0.001
Ferrous sulphate	0.001
Manganous chloride	0.002
Menadione	0.001
Cyanocobalamin	0.001
L-Cysteine hydrochloride	0.020
L-Tryptophan	0.020
Pyridoxine hydrochloride	0.003
Calcium pantothenate	0.003
Nicotinamide	0.003
Biotin	0.0003
Thiamine hydrochloride	0.00004
Adenine	0.010
Guanine	0.010
Xanthine	0.010
Uracil	0.010
Agar	8.000
Final pH (at 25°C)	7.4±0.2

**Formula adjusted, standardized to suit performance parameters

Directions

Suspend 31.4 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. Mix well and pour into sterile Petri plates.

Principle And Interpretation

The goal of an antimicrobial susceptibility test is to predict through an in vitro assessment the likelihood of successfully treating an infection with a particular antimicrobial agent. There are several continual or novel methods for performing antibacterial susceptibility testing. These include the disk diffusion test, broth microdilution, agar gradient and rapid automated instrument methods (1). Hi-Sensitivity™ Test HiVeg™ Agar, which is used for antimicrobial susceptibility tests, is a semi-defined medium in which the mineral contents have been stabilized to give reproducible results. The thiamine and thymidine content is very low thus making it most suitable for testing antimicrobial activity of sulfonamide. However some mutant strains which are totally dependent on thiamine and thymidine for their growth, will not grow on Hi-Sensitivity™ Test HiVeg™ Agar, due to very low levels of these compounds in the media as they are the naturally occurring antagonist of trimethoprim. These strains should be carefully recognized (2,3,4).

Hi-Sensitivity™ Test HiVeg™ Agar has been so designed to overcome the problems occurring in Mueller-Hinton Media that are as follows (5,6,7,8,9,10,11).

Mueller Hinton Agar and Mueller Hinton Broth give different MIC values.
Mueller Hinton Agar shows antagonistic effect towards tetracycline.
High levels of sulfonamide and trimethoprim antagonists.
Media prepared using peptone of different manufacturers give poor reproducibility.
Poor growth supporting ability for Streptococci and variable growth rates with gram-positive organisms.

Some pathogenic organisms are nutritionally dependent due to their intrinsic demands for special growth factors. Supplemental nutrients can be added to Hi-Sensitivity Test Agar to improve the growth of these organisms (12).

The following nutrients can be used.

Nutrient	Microorganism(s)
1.Laked blood (5% v/v)	Neisseria and Streptococci
2.Fildes Peptic Digest of Blood (5% v/v)	Haemophilus
3.Menadione (0.5 mcg/ml) and Thiamine hydrochloride (2 mcg/ml)	Dwarf colonies of Staphylococcus aureus and coliform bacteria.
4.Pyridoxine hydrochloride	Symbiotic Streptococci (1mcg/ml)

Supplementation of Hi-Sensitivity™ Test HiVeg™ Agar with 10% horse blood can be used for susceptibility testing of Helicobacter pylori (13).

Hi-Sensitivity™ Test HiVeg™ Agar is prepared by completely replacing animal based peptone with vegetable peptones to avoid BSE/TSE risks associate with animal peptones. HiVeg™ hydrolysate, HiVeg™ peptone provides nitrogenous and carbonaceous compounds, long chain amino acids and other essential growth nutrients. Dextrose is the energy source. Vitamins provides nitrogen, carbon compounds and other essential growth nutrients.

Type of specimen

samples

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 clinical specimens. Safety guidelines may be referred in individual safety data sheets.

Limitations

This medium is recommended for susceptibility testing of pure cultures only.
Inoculum density may affect the zone size. Heavy inoculum may result in smaller zones or too less inoculum may result in bigger zones.
Fastidious organisms may not grow on this medium and may require supplementation of blood.
However some mutant strains which are totally dependent on thiamine and thymidine for their growth, will not grow on Hi-Sensitivity™ Test HiVeg™ Agar, due to very low levels of thiamine and thymidine these compounds in the media as they are the naturally occurring antagonist of trimethoprim. These strains should be carefully recognized (2,12,14).
As antimicrobial susceptibility is carried with antibiotic disc, proper storage of the disc is desired which may affect the potency of the disc.
Under certain circumstances, the in vitro results of antibiotic susceptibility may not show the same in vivo.

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 0.8% Agar gel.

Colour and Clarity of prepared medium
Basal medium: Light amber coloured, clear to slightly opalescent gel. After addition of 5%v/v laked blood: Red to chocolate coloured, opaque gel forms in Petri plates.

Reaction

Reaction of 3.14% w/v aqueous solution at 25°C. pH: 7.4±0.2

pH

7.20-7.60

Cultural Response

Cultural characteristics observed with added 5%w/v laked blood, after an incubation at 35 - 37°C for 18 - 24 hours .

Organism (ATCC)	Inoculum (CFU)	Growth	Recovery
^Bacillus spizizenni ATCC 6633 (00003*)	50-100	good-luxuriant	>=70%
**Phocaeicola vulgatus ATCC 8482	50-100	good-luxuriant	>=70%
Enterococcus faecalis ATCC 29212 (00087*)	50-100	good-luxuriant	>=70%
Salmonella Typhimurium ATCC 14028 (00031*)	50-100	good-luxuriant	>=70%
Staphylococcus aureus subsp. aureus ATCC 25923 (00034*)	50-100	good-luxuriant	>=70%
Streptococcus pyogenes ATCC 19615	50-100	good-luxuriant	>=70%

*- Corresponding WDCM numbers (^) Formerly known as Bacillus subtilis subsp. spizizenii (**)Formerly known as Bacteroides vulgatus

Storage and Shelf Life

Store dehydrated and the prepared medium at 2-8°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 (1,14).

Product Information

More Information
Product Name	Hi-Sensitivity™ Test HiVeg™ Agar
SKU	MV485
Product Type	HiVeg™
Physical Form	Powder
Origin	Animal Free (Veg)
Packaging type	HDPE
References	1.Isenberg, H.D. Clinical Microbiology procedures Handbook. 2nd Edition2.Tanner E. I. and Bullin C. H., 1974, J. Clin. Path., 27:565. 3.Thomas M. and Bond L., 1973, Med. Lab. Technol., 30:277. 4.Barker J., Healing D., and Hutchinson J. G. P., 1972, J. Clin. Path., 25:10865.Ericsson H. M. and Sherris J. C., 1971, Acta. Pathol. Microbiol Scand Suppl., 217:1. 6.Garrod L. P. and Waterworth P. M., 1971, J. Clin. Path., 24:779. 7.Reller L. B., Schoenknecbt F. D., Kenny M. A. and Sherris J. C., 1974, J. Infect. Dis., 130:454. 8.Duncan I. B. R., 1974, Antimicrob. Agents Chemother., 5:9.9.Yourassowsky E., Vanderlinden M. P. and Schoutens E., 1974, J. Clin. Path., 27:897.10.Neussil H., 1976, Chemother., Vol. 2 : 33. 11.Bridson E.Y., 1976, Arztl. Lab., 22:373. 12.Acar J. F., 1980, Antibiotics in Laboratory Medicine, Lorian V. (Ed.), Williams and Wilkins, Baltimore, USA, 48-51. 13.Hartzen S.H., Anderson L.P., Bremmelgaard A. et.al, 1997, Antimicrob. Agents Chemother., 41.2634-2639. 14.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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