CHROMOCULT COLIFORM AGAR ES (SELETIVIDADE AUMENTADA) MERCK - EMB 500G
Modelo: 1008500500
-
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Descrição
CHROMOCULT COLIFORM AGAR ES (SELETIVIDADE AUMENTADA) MERCK
APROVADO PELA AOAC LICENÇA N° 041002
RENDIMENTO 34,5g/L
Embalagem 500g.
Typical Composition (g/litre)
Peptone 5.0; potassium chloride 7.5; MOPS 10.0; bile salts 1.15; propionate 0.5; Agar-Agar 10.0; 6-Chloro-3-
indoxyl-beta-Dgalactopyranoside 0.15; isopropyl-beta-D-thiogalactopyranoside 0.1; 5-bromo-4-chloro-3-indoxyl-
beta-D-glucuronic acid 0.1.
Preparation
Suspend 34.5 g in 1000 ml of purified water and heat to boiling with frequent agitation until completely dissolved (approximately 45 minutes).
Do not autoclave, do not overheat.
Immediately cool the medium in a water bath at 45-50°C (a precipitate appears if a period of 2 hours is exceeded).
pH: 7.0 ± 0.2 bei 25°C
The medium is clear and colourless.
Sample Preparation
Prepare test samples using standard laboratory techniques such as those described in the Bacteriological
Analytical Manual or the ISO standard specific for the product concerned.
To minimize possible interference between the coloration of coliforms/ E. coli and the sample (e.g. low pH), it is
advisable to dilute the sample 1:10 in a buffered solution (e.g. add 450 ml Butterfield's phosphate buffer, buffered peptone water, or buffered sodium chloride peptone broth to blender jar containing 50 g of sample and blend 2 min) and then to perform further dilutions as needed.
Application
Chromocult® Coliform Agar ES is usually inoculated by the pour plate method.
Using a sterile pipette, transfer 1 ml of the liquid test sample (or 1 ml from the appropriate dilution) to a sterile Petri dish.
Pour into each Petri dish approximately 15 ml of the Chromocult® Coliform Agar while the medium is still liquefied,
but only after the medium has been cooled to 45-50°C in a water bath. Carefully swirl the plate until the inoculum is thoroughly mixed with the medium. Allow the mixture to solidify on a cool horizontal surface.
Incubate the inoculated dishes aerobically at 35-37°C in an inverted position (agar side up) for 24 hours.
After incubation, examine the plates for the presence of typical colonies of E. coli and other coliforms.
Results
Count the dark blue to violet colonies as E. coli and the salmon to red colonies as other coliforms.
The total of all red and blue colonies is the total coliform count.
Some E. coli (3-4%) are ß-glucuronidase-negative and grow as salmon-red colonies, e.g. E. coli O157 strains.
Accompanying flora appear as colorless colonies, except for some organisms, which possess ß-D-glucuronidase
activity. These colonies appear light blue to turquoise in color.
The total coliform count should not exceed 150 typical CFU and 300 total CFU (total coliforms and accompanying bacteria) per plate. Above these levels, the colonies cannot be counted accurately. Samples, which are expected to exceed these maximum levels, should be diluted prior to inoculation.
APROVADO PELA AOAC LICENÇA N° 041002
RENDIMENTO 34,5g/L
Embalagem 500g.
Typical Composition (g/litre)
Peptone 5.0; potassium chloride 7.5; MOPS 10.0; bile salts 1.15; propionate 0.5; Agar-Agar 10.0; 6-Chloro-3-
indoxyl-beta-Dgalactopyranoside 0.15; isopropyl-beta-D-thiogalactopyranoside 0.1; 5-bromo-4-chloro-3-indoxyl-
beta-D-glucuronic acid 0.1.
Preparation
Suspend 34.5 g in 1000 ml of purified water and heat to boiling with frequent agitation until completely dissolved (approximately 45 minutes).
Do not autoclave, do not overheat.
Immediately cool the medium in a water bath at 45-50°C (a precipitate appears if a period of 2 hours is exceeded).
pH: 7.0 ± 0.2 bei 25°C
The medium is clear and colourless.
Sample Preparation
Prepare test samples using standard laboratory techniques such as those described in the Bacteriological
Analytical Manual or the ISO standard specific for the product concerned.
To minimize possible interference between the coloration of coliforms/ E. coli and the sample (e.g. low pH), it is
advisable to dilute the sample 1:10 in a buffered solution (e.g. add 450 ml Butterfield's phosphate buffer, buffered peptone water, or buffered sodium chloride peptone broth to blender jar containing 50 g of sample and blend 2 min) and then to perform further dilutions as needed.
Application
Chromocult® Coliform Agar ES is usually inoculated by the pour plate method.
Using a sterile pipette, transfer 1 ml of the liquid test sample (or 1 ml from the appropriate dilution) to a sterile Petri dish.
Pour into each Petri dish approximately 15 ml of the Chromocult® Coliform Agar while the medium is still liquefied,
but only after the medium has been cooled to 45-50°C in a water bath. Carefully swirl the plate until the inoculum is thoroughly mixed with the medium. Allow the mixture to solidify on a cool horizontal surface.
Incubate the inoculated dishes aerobically at 35-37°C in an inverted position (agar side up) for 24 hours.
After incubation, examine the plates for the presence of typical colonies of E. coli and other coliforms.
Results
Count the dark blue to violet colonies as E. coli and the salmon to red colonies as other coliforms.
The total of all red and blue colonies is the total coliform count.
Some E. coli (3-4%) are ß-glucuronidase-negative and grow as salmon-red colonies, e.g. E. coli O157 strains.
Accompanying flora appear as colorless colonies, except for some organisms, which possess ß-D-glucuronidase
activity. These colonies appear light blue to turquoise in color.
The total coliform count should not exceed 150 typical CFU and 300 total CFU (total coliforms and accompanying bacteria) per plate. Above these levels, the colonies cannot be counted accurately. Samples, which are expected to exceed these maximum levels, should be diluted prior to inoculation.
