Microbiological Testing of Water using Readycult Coliforms and Enterococci

1.01295.0001 1.01298.0001

Readycult Coliforms 50 Readycult Coliforms 100

Contents: 20 snap pack. 1 snap pack each for 50 ml or 100 ml of water sample.
s Application Selective enrichment broth for the simultaneous detection of total coliforms and E.coli within the bacteriological water examination.

s Disposal Autoclave the broth (15 min./121 C). Alternatively heat the broth for 30 min. in boiling water or use a proper disinfectant (e.g. Extran MA 04).

1.01299.0001

Readycult Enterococci 100

Contents: 20 snap pack.

1 snap pack each for 100 ml of water sample. s Application Selective enrichment broth for the detection of Enterococci and D-Streptococci within the bacteriological water examination. s Principle The peptone mixture guarantees rapid growth of Enterococci. Sodium-azide largely inhibits the accompanying flora, especially the gram-negative. The chromogenic substrate X-GLU is cleaved, stimulated by selected peptones, by the enzyme -D-Glucosidase present in Enterococci. This results in an intensive blue-green colour change of the broth. s Composition in g/blister Peptones 0.86; sodium chloride 0,64; sodium-azide 0,06; X-GLU 0.004; Tween 80 0.22. s Procedure 1. Add 100 ml of water sample into a sterile, transparent 250 ml vessel with screw cap. In case the sample is to be stored below +25 C the examination has to be started within 6 hours. Exceptionally the sample can be stored at +2 to + 8 C (refrigeration) for up to 24 hours. 2. Take out one snap pack, shortly tap to ensure the granules at the bottom of the snap pack. Bend the upper part of the snap pack until it breaks open. Attention: To avoid contamination risk do not touch the opening! 3. Add the contents to the water sample. Seal the vessel and shake to completely dissolve the granules. 4. Incubation: 18 24 h at 35 to 37 C. If incubated at room temperature (+20 to +25 C) the incubation time is prolonged to 48 hours. s Interpretation of Results Negative: No colour change. The broth remains slightly yellow. Positive: Any colour change of the broth to blue-green, even only in the upper section of the broth, confirms presence of Enterococci (X-GLU reaction). No decolouration with shaking! s Disposal Autoclave the broth (15 min./121 C). Alternatively heat the broth for 30 min. in boiling water or use a proper disinfectant (e.g. Extran MA 04). s Additives Merck Cat.No. 1.13203.0001 1.09293.0100 1.07551.2000 1.13311.0001 Product UV-Lamp 366 nm KOVACS Indole Reagent Extran MA 04 disinfectant

Rapid detection and identification of microorganisms are of high importance. The use of fluorogenic and chromogenic substrates, utilizing specific enzymatic activities of certain microorganisms, for rapid and sensitive detection of bacteria has proved to be a powerful alternative to conventional methods:

Microbiological Analysis of Water

Simple ~ Rapid ~ Low cost
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s Principle The high nutritional quality of the peptones and the incorporated phosphate buffer guarantee rapid growth of coliforms whereas laurylsulfate largely inhibits the accompanying flora, especially the gram-positive. By adding the chromogenic substrate X-GAL which is cleaved by coliforms and the fluorogenic substrate MUG which is highly specific for E.coli the simultaneous detection of total coliforms and E.coli is possible. The presence of total coliforms is indicated by a blue-green colour of the broth and E.coli by a blue fluorescence under UV-light. s Composition in g/blister Tryptose 0.25 (50)/0.5 (100); sodium chloride 0.25/ 0.5; sorbitol 0.05/0.1; tryptophan 0.05/0.1; di-potassium hydrogen phosphate 0.135/0.27; potassium dihydrogen phosphate 0.1/0.2; Laurylsulfate sodium salt 0,005/0.01; X-GAL 0.004/0.008; MUG 0.0025/0.005; IPTG 0.005/0.01. s Procedure 1. Add 50/100 ml of water sample into a sterile, transparent 100/250 ml vessel with screw cap. Attention: Please use material e.g. glass that is none fluorescing itself! In case the sample is to be stored below +25 C, the examination has to be started within 6 hours. Exceptionally the sample can be stored at +2 to +8 C (refrigeration) for up to 24 hours. 2. Take out one snap pack, shortly tap to ensure the granules at the bottom of the snap pack. Bend the upper part of the snap pack until it breaks open. Attention: To avoid contamination risk do not touch the opening! 3. Add the contents to the water sample. Seal the vessel and shake to completely dissolve the granules. 4. Incubation: 18 24 h at 35 to 37 C. If incubated at room temperature (+20 to +25 C) the incubation time is prolonged to 48 hours. s Interpretation of Results for the detection of Total

Convenient

The media are supplied in ready-to-use snap pack format Designed for on-site testing No media preparation required Simply add medium, incubate and read results Results within 18 to 24 hours results 3 4 days earlier than traditional methods Hands-on time in less than 1 minute Simple detection by blue-green colour/ fluorescence Colour comparison charts not needed No interpretation problems with water of high turbidity Reduction of workload and material costs: - Only 1 tube - Durham-tubes not required Enzyme substrate reaction specifically targeted

Rapid

Easy to read

Coliforms/E.coli:
Negative: No colour change. The broth remains slightly yellow. Total coliforms: Any colour change of the broth to bluegreen, even only in the upper section of the broth, confirms presence of coliforms ( X-GAL reaction ). No decolouration with shaking! E.coli: Check blue-green coloured vessels for fluorescence by using UV-lamp in front of the vessel. Light blue fluorescence indicates presence of E.coli ( MUG reaction ). Attention: Protect your eyes from direct UV light! To confirm E.coli in the vessel with positive fluorescence overlay the broth with 2.5 ml of KOVACs reagent (Indole reaction). A red ring confirms presence of E.coli. Colour change to blue-green Total coliforms E.coli Negative + + yellow colour + + Fluorescence IndoleReaction

Economical

Pack size 1 each 100 ml bottle 2 litres

High performance

CULTURA Mini-Incubator 220 235 Volt CULTURA Mini-Incubator 100 110 Volt

1 each

1.15533.0001

1 each

Merck KGaA - 64271 Darmstadt - Germany Fax: ++49-61 51/72 90 06 2 e-mail: environ.analysis@merck.de or your local Merck (on Internet http://www.merck) or Merck dealer

The innovative enrichment media for detection of Enterococci, total coliforms and E.coli in water

The colour makes the difference!

Readycult Coliforms Readycult Enterococci

Readycult Coliforms

The combination of a chromogenic and fluorogenic substrate is used for the simultaneous detection of total coliforms and E.coli. Differentiation between total coliforms and other gram-negatives: total coliforms produce a blue-green colour after hydrolisation of X-GAL by the characteristic enzyme b-D-galactosidase. Unequivocal identification of E.coli with fluorescence due to metabolisation of MUG by the enzyme b-D-glucuronidase.

1. Add Readycult to 50 or 100 ml sample and mix

2. Incubate 18 24 h at 35 37 C

1. Add Readycult to 100 ml sample and mix

2. Incubate 18 24 h at 35 37 C

3. Interpretation of results

3. Interpretation of results

Readycult Enterococci

The chromogenic substrate X-GLU is used to differentiate Enterococci from other bacteria. The medium turns to blue-green colour, when the substrate X-GLU is cleaved by the enzyme b-D-Glucosidase, characteristic for Enterococci.

Positive colour change to blue-green

Negative no colour change to blue-green

Positive 1 colour change to blue-green 2 Fluorescence + 3 Indole

Positive colour change to blue-green

Negative no colour change to blue-green

Presence of Total Coliforms

No Coliforms

Presence of E. coli

Presence of Enterococci

No Enterococci