Catalase Test Procedure, Principle, Precautions & Results

Catalase is the enzyme that catalyzes to split up hydrogen peroxide into its reactant (water and oxygen). It has produced by a special kind of bacteria called staphylococci. Catalase enzyme has been utilizing in many different works like, topical disinfectant in wounds, bubbling that is observed because of the evolution of oxygen gas.

catalase structure

Uses of Hydrogen peroxide:

  1. It is the strong oxidizing agent, help in industries to oxidize the other compound or element.
  2. It acts as a natural anti-septic, that is why it is used to clear wounds and infection.
  3. It acts well in preventing the tooth from cavities.
  4. Having an edge in beauty products.
  5. With using hydrogen peroxide, we can improve the seed germination.
  6. Act as a good De-Funk Musty Fabric.
  7. Spray few drops of hydrogen peroxide onto light-colored carpets or rugs in order to clean stains from mud, food, etc. Keep in your mind that hydrogen peroxide will bleach some fabrics, so try this technique first in an inconspicuous fabric.
  8. Shake the fruits and veggies with hydrogen peroxide and let soak for a while, before washing them all in clean water to remove the contaminants.
  9. Mix equal ratio of hydrogen peroxide and vinegar in a bottle having a dropper. Put several drops in each ear before to take a swim in order to prevent any infection.
  10. Steep your toothbrushes in hydrogen peroxide to make free from staph bacteria and other related germs common to the bathroom environment.
  11. Mix equal ratio of hydrogen peroxide and warm water to make a foot soak that will naturally soften corns and calluses.
  12. Spray few drops of dilute hydrogen peroxide into children’s lunchboxes or cooler. Left it for a while, then wash it to clean and disinfect.
  13. Add the few drops of hydrogen peroxide over wettish hair and let it remain in for an hour, before rinsing out. It will give your hair a shine, without blow your hair.
  14. Splash few drops of dilute hydrogen peroxide (because concentrated can damage the food) on cutting boards to extinguish germs from any pouring foods that could have the ability to absorb the drops into the surface of the board.
  15. Add some hydrogen peroxide in the dropper bottle and sprinkle few drops of it into dirty rags and cleaning sponges and let them in it for about a half hour to disinfect.

catalase test procedure

Principle of Catalase Test:

The enzyme catalase helps in the breakdown of hydrogen peroxide into its reactants. The presence of the enzyme in a midgut bacterial culture (keep in the mind that the culture must not be older than 24 hours), is must at a point when to introduce a small amount of inoculum bacterial isolate into hydrogen peroxide, as a result the vigorous elaboration of oxygen bubbles observes. The lack of catalase may responsible for a lack of or weak bubble production.

Catalase test purpose:

  1. The morphologically the members of the genus Enterococcus  Streptococcus (catalase negative) and the members of the genera Staphylococcus (catalase positive) can be distinguished by using the catalase test.
  2. Catalase test has been used for many years as an aid to the identification of the genera Enterobacteriaceae.
  3. The catalase test, using 15% H2O2 is used to distinguish between aerotolerant species of Clostridium, from Bacillus species.
  4. Catalase test is as well utilized for the reorganization of Mycobacterium tuberculosis.

Catalase Test Procedure – How to do Catalase test:

  • Put a minute amount of growth from your culture onto a microscope slide (make sure it is cleaned). However, if you are using colonies from a blood agar plate, so make sure not to scrape up any of the blood agar, as red blood cells are catalase positive and any contaminating agar may give a false positive test.
  • After which add a few drops of hydrogen peroxide onto the smear. If needed, mix the both with the help of the toothpick. Make sure not to use a metal loop or needle with hydrogen peroxide; it may give a false positive test or degrade the metal.
  • If the test is positive than the rapid evolution of O2 in the form of bubbling would observed.
  • If the test is being negative than there are no bubbling or only a few disperse bubbles would be observed.
  • Dispose of your slide in the biohazard glass disposal container (a type of container mostly using for this purpose). Dispose of any toothpicks in the Pipet Keeper.

Precautions of Catalase Test:

  1. Use fresh and unstable hydrogen peroxide.
  2. Don’t be used Iron wire, during the reaction.
  3. Don’t add organism to the reagent, may cause a false positive result.
  4. Some bacteria produce a per-oxidase, result in weak positive result.

Relevant tests:

Oxidase test:

This is a test to figure out if an organism performs aerobic respiration. It is a check for the existence of the electron transport chain that is the last phase of aerobic respiration. Typically, oxygen is the endmost electron acceptor for this system. In the oxidase test, an artificially designed final electron acceptor (N,N,N’,N’-tetramethyl-phenylenediamine dihydrochloride) has been used in the replacement of oxygen. This acceptor is a chemical dye that gradually changes color to a dark blue or purple when it takes the electron from the last element (cytochrome oxidase) in the electron transport chain.

How to do oxidase test:

  • To open a brand new reagent dispenser: Hold reagent dropper upright and point tip away from yourself it will create a room between both. Grasp the middle with the help of thumb and forefinger and squeeze politely to crush the glass ampule inside the dropper. After which tap the bottom on the tabletop a few times.
  • Place one drop of reagent onto the culture on the swab to make it moist.
  • Positive reactions turn the bacteria violet to purple, within no time.
  • Delayed reactions should be ignored.

Hi, This is Hamza Khan from Peshawar, Pakistan. I am a 2nd prof MBBS Student at Bannu Medical College and a hobby Blogger. The Purpose of this site is to share my knowledge and Guide new Medical Students.

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