This agar is used to identify organisms that are capable of producing the enzyme lipase. This enzyme is secreted and hydrolyzes triglycerides to glycerol and three long chain fatty acids. These compounds are small enough to pass through the bacterial cell wall.
What is the function of spirit blue dye in spirit blue agar?
Spirit blue or Opal blue SS is a dye with formula C37H30ClN3. It is used as an indicator in agar for detection of lipase activity in bacteria.
How do you make spirit blue agar?
A satisfactory emulsion can be prepared by dis- solving 10 g gum acacia or 1 mL polysorbate 80 in 400 mL warm purified water, adding 100 mL cottonseed or olive oil and agitating vigorously to emulsify. Spirit Blue Agar contains peptone as a source of carbon, nitrogen, vitamins and minerals.
How do you make Tributyrin?
Dissolve 20 g in 1 litre distilled water. Sterilize by autoclaving at 121°C for 15 minutes. Let cool to 80°C and add 10g neutral Tributyrin (91010). Mix thoroughly to emulsify the Tributyrin completely.
What type of medium is Spirit Blue agar?
Spirit blue agar is a nutrient medium with the dye spirit blue present. It also contains an emulsified lipid to serve as the target for the enzyme. For the test in this program, that emulsified lipid is assumed to be corn oil.
Why do bacteria hydrolysis lipids?
Lipids generally are nonpolar molecules that do not dissolve well in water. In order to utilize fats, bacterial cells secrete exoenzymes known as lipases outside of the cell that hydrolyze the lipid to fatty acids and glycerol. These bacteria capable of producing exoenzyme lipase are called lipolytic bacteria.
What type of media is Spirit Blue Agar?
What Colour is Spirit Blue?
The hexadecimal color code #514f72 is a medium dark shade of blue-magenta. In the RGB color model #514f72 is comprised of 31.76% red, 30.98% green and 44.71% blue.
What is starch agar?
Starch agar is a differential medium that tests the ability of an organism to produce the extracellular enzymes (exoenzymes) α-amylase and oligo-1,6-glucosidase that are secreted out of the bacteria and diffuse into the starch agar.
What is Tributyrin Agar?
TRIBUTYRIN AGAR is a medium used for the detection and enumeration of lipolytic bacteria and moulds in foodstuffs and other materials.
What is Tributyrin used for?
Tributyrin is a triglyceride naturally present in butter. It is an ester composed of butyric acid and glycerol. Among other things, it is used as an ingredient in making margarine. It is present in butter and can be described as a liquid fat with an acrid taste.
Which media is used in lipid hydrolysis test?
In this experimental procedure, tributyrin agar is used to demonstrate the hydrolytic activities of the exoenzyme lipase. The medium is composed of nutrient agar supplemented with the triglyceride tributyrin as the lipid substrate.
What is Spirit blue agar used for?
Spirit Blue Agar is for use with Lipase Reagent or other lipid source for detecting and enumerating lipolytic microorganisms.
How do you test for Gram positive bacteria on blood agar?
The bacterium from the pure culture was pulled and streaked with an inoculating loop on the blood agar, then the plate was left in the incubator at 37 °C for 48 hours. The result was observed and recorded. Spirit Blue Agar was the final test for Gram (+) bacteria.
What is the color of the Orange colony on MacConkey agar?
The orange colony was Gram stained, observed, and recorded. Under the light microscope the colony appeared to be pink and purple color, meaning the culture was still mixed. Since the Gram (+) bacteria had been isolated, MacConKey Agar (MAC) and Eosin Methylene Blue (EMB) tests were selected in order to isolate the Gram (-) bacteria.
What is strengstreak isolation on nutrient plate agar?
Streak isolation on a nutrient plate agar was performed to separate a mixture of bacteria to pure culture. The nutrient agar was then incubated at 37°C for 48 hours. The isolation plate was observed and recorded. The plates showed two colonies with distinguishing colors; a white colored colony and an orange colored colony.
