PHENOL RED CARBOHYDRATE TEST MEDIA

Cat. no. Y301 Phenol Red Broth with Adonitol, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y302 Phenol Red Broth with Arabinose, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y304 Phenol Red Broth with Dextrose, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y305 Phenol Red Broth with Dulcitol, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y306 Phenol Red Broth with Galactose, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y321 Phenol Red Broth with Glucoside, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y322 Phenol Red Broth with Glycerol, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y307 Phenol Red Broth with Inositol, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y308 Phenol Red Broth with Inulin, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y309 Phenol Red Broth with Lactose, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y310 Phenol Red Broth with Maltose, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y311 Phenol Red Broth with Mannitol, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y319 Phenol Red Broth with Mannose, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y312 Phenol Red Broth with Raffinose, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y313 Phenol Red Broth with Rhamnose, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y314 Phenol Red Broth with Salicin, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y315 Phenol Red Broth with Sorbitol, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y316 Phenol Red Broth with Sucrose, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y317 Phenol Red Broth with Trehalose, 16x125mm Tube, 10ml 20 tubes/box
Cat. no. Y318 Phenol Red Broth with Xylose, 16x125mm Tube, 10ml 20 tubes/box
Note: Each Phenol Red Broth tube listed above includes a durham tube.

INTENDED USE

Hardy Diagnostics Phenol Red Carbohydrate Test Media is intended to be used in the differentiation of microorganisms based on fermentation reactions.

SUMMARY

Phenol Red Carbohydrate Test Media contains pancreatic digest of casein, sodium chloride, a carbohydrate, and phenol red. Pancreatic digest of casein provides the nutrients and growth factors necessary for growth. This ingredient is low in carbohydrates that may interfere with the fermentation reactions of the media. The individual carbohydrates are added at a concentration of 1%. Phenol red is the acid-base indicator in the media. It is red at an alkaline pH, and shifts to yellow when acid is produced during carbohydrate fermentation. Durham tubes, also known as fermentation tubes, are added to visualize gas production produced during carbohydrate fermentation reactions. After incubation, yellow media with gas trapped in the durham tube is indicative of positive acid and gas fermentation reactions.

Phenol Red Broth Base lacks a carbohydrate. It has a fill of nine milliliters so that one milliliter of a carbohydrate solution can be added aseptically to the tube. Phenol Red Broth Base may also be used in conjunction with Phenol Red Broth containing carbohydrate as a negative control.

FORMULA

Ingredients per liter of deionized water:*

Pancreatic Digest of Casein 10.0gm
Sodium Chloride 5.0gm
Phenol Red 18.0mg

Carbohydrates are added at a concentration of 10.0gm/L.

Final pH 7.4 +/- 0.2 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-8ºC. away from direct light. Media should not be used if there are any signs of contamination, deterioration, discoloration, or if the expiration date has passed. Product is light and temperature sensitive.

PRECAUTIONS

PROCEDURE

Inoculate the Phenol Red Broth, with added carbohydrate of choice, with a pure culture of the organism. Incubate at 35-37ºC., and examine daily for up to 7 days.

Use an uninoculated tube of Phenol Red Broth of the same carbohydrate as a negative control.

INTERPRETATION OF RESULTS

Carbohydrate fermentation can be measured using acid and/or gas production. Carbohydrate acid fermentation is indicated by the appearance of a yellow color in the broth. A gas fermentation reaction is recorded when gas is observed within the durham tube for up to seven days following inoculation. A negative result is noted when the medium remains red and no gas is produced in the durham tube. A negative reaction will appear the same as an uninoculated control tube.

LIMITATIONS

The reactions observed from this media are not sufficient to speciate an organism. This product is not intended for primary isolation of patient specimens. This product is used in conjunction with other biochemical and serological tests to identify cultures of isolated organism. (1,4)

For fastidious organisms, such as some Streptococcus spp., Purple Broth with Carbohydrates (Cat. no. Y100-Y119) is recommended, as it provides a more nutritionally rich media for organism growth.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, other culture media, swabs, applicator sticks, incinerators, and incubators, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Phenol Red Broth with Adonitol:
Enterobacter aerogenes
ATCC ® 13048
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Escherichia coli
ATCC ® 25922
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Arabinose:
Enterobacter aerogenes
ATCC ® 13048
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Serratia marcescens
ATCC ® 8100
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Dextrose:
Escherichia coli
ATCC ® 25922
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Branhamella ( Moraxella )
catarrhalis
ATCC ® 25240

A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Dulcitol:
Salmonella paratyphi
ATCC ® 9150
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Serratia marcescens
ATCC ® 8100
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Galactose:
Escherichia coli
ATCC ® 25922
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Serratia marcescens
ATCC ® 8100
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Glucoside:
Klebsiella pneumoniae
ATCC ® 13883
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Edwardsiella tarda
ATCC ® 15947
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Glycerol:
Citrobacter freundii
ATCC ® 8090
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Salmonella enterica
ATCC ® 14028
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Inositol:
Enterobacter aerogenes
ATCC ® 13048
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Escherichia coli
ATCC ® 25922
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Inulin:
Streptococcus mutans
ATCC ® 25175
D 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas negative


Streptococcus mitis
ATCC ® 6249
D 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Lactose:
Escherichia coli
ATCC ® 25922
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Proteus mirabilis
ATCC ® 12453
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Maltose:
Escherichia coli
ATCC ® 25922
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Proteus mirabilis
ATCC ® 12453
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Mannitol:
Escherichia coli
ATCC ® 25922
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Proteus mirabilis
ATCC ® 12453
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Mannose:
Klebsiella pneumoniae
ATCC ® 13883
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Proteus mirabilis
ATCC ® 12453
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Raffinose:
Enterobacter aerogenes
ATCC ® 13048
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Serratia marcescens
ATCC ® 8100
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Rhamnose:
Enterobacter aerogenes
ATCC ® 13048
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Serratia marcescens
ATCC ® 8100
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Salicin:
Enterobacter aerogenes
ATCC ® 13048
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Salmonella enterica
ATCC ® 14028
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Sorbitol:
Escherichia coli
ATCC ® 25922
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Proteus mirabilis
ATCC ® 12453
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Sucrose:
Enterobacter aerogenes
ATCC ® 13048
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Salmonella enterica
ATCC ® 14028
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Trehalose:
Klebsiella pneumoniae
ATCC ® 13883
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Morganella morganii
ATCC ® 25830
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


Phenol Red Broth with Xylose:
Klebsiella pneumoniae
ATCC ® 13883
A 18-48hr 35°C Aerobic Growth
Positive Reaction:
yellow color change,
gas positive


Morganella morganii
ATCC ® 25830
A 18-48hr 35°C Aerobic Growth
Negative Reaction:
no yellow color change,
gas negative


User Quality Control

PHYSICAL APPEARANCE

E. coli growing in Phenol Red Broth with Adonitol

Escherichia coli (ATCC ® 13048) growing in Phenol Red Broth with Adonitol (Cat. no. Y301). Incubated aerobically for 24 hours at 35ºC. Growth and a yellow color change was indicative as positive for adonitol fermentation. The bubble in the durham tube was indicative of gas production.

Phenol Red Broth with Adonitol

Uninoculated tube of Phenol Red Broth with Adonitol (Cat. no. Y301).

REFERENCES

1. Jorgensen., et al. Manual of Clinical Microbiology, American Society for Microbiology, Washington, D.C.

2. Tille, P., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

3. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.

4. Koneman, E.W., et al. Color Atlas and Textbook of Diagnostic Microbiology, J.B. Lippincott Company, Philadelphia, PA.

5. MacFaddin, J.F. Biochemical Tests for Identification of Medical Bacteria,, Lipincott Williams & Wilkins, Philadelphia, PA.

6. Quality Assurance for Commercially Prepared Microbiological Culture Media, M22. Clinical and Laboratory Standards Institute (CLSI - formerly NCCLS), Wayne, PA.


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