Agar slants are inoculated with dark centered

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- Agar Slants are inoculated with Dark-Centered Colonies • Incubated at 35° for 24 ± 2 Hours • Presence of Gram Negative, Non-Spore-Forming Rods is Positive - The Presence of Gram Negative, Non-Spore-Forming Rods that produce Gas and Ferment Lactic Acid constitutes a Positive Completed Test All Samples involved in the Multiple Tube Fermentation should test Negative. Any Positive Results indicate a serious Breakdown in Water Treatment, usually involving Chlorination Problems or Water Main Breaks (an increasing Problem with this Country ʼ s aging Infrastructure).
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Water and Sewage Treatment Page 3 Membrane Filtration Method • 100 ml of Sample Water is drawn through a 0.45 m m Sterile Membrane Filter • The Membrane Filter is then incubated on a Pad saturated with Growth Medium • Any Colonies that grow can be further tested The Presence of more than 1 Colony generally indicates that the Water is not Safe to drink. ONPG and MUG Test • Water Sample inoculated into Nutrient Broth containing: - ONPG ( o -Nitrophehyl-b- D Galactopyranoside) • ONPG is a Colorimetric (G = measure with color) Compound that will produce a Yellow Product when its broken down by the Enzyme b -Galactosidase in Coliforms. - MUG (4-Methylumbelliferyl- b - D -Glucuronide) • MUG is a Fluorogenic (G = produce fluorescence) Compound that will produce a Blue Fluorescent Compound when broken down by the Enzyme b -Glucuronidase in Coliforms • Within 24 Hours any Coliforms that grow will produce the Enzymes b-Galactosidase and b-Glucuronidase - The ONPG Test can indicate the Presence of Coliforms by demonstrating the Presence of b -Galactosidase
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