preview

Determining Unknown in Microbiology Lab

Decent Essays

Determining an Unknown Through Deferential Stains and Biochemical Tests Introduction There are many reasons for knowing the identity of microorganisms. The reasons range from knowing the causative agent of a disease in a patient, so as to know how it can be treated, to knowing the correct microorganism to be used for making certain foods or antibiotics. This study was done by applying all of the methods that have been learned so far in the microbiology laboratory class for the identification of unknown bacteria. The identification process can be completed with a series of deferential stains and biochemical tests. Creating a dichotomous key helps to limit the amount of biochemical tests done on an unknown organism and by observation …show more content…

The enzyme urease breaks urea down into NH3 and CO2. An orange broth containing urea is used for this test and needs to be inoculated with the gram negative bacteria. A pink color in the medium indicates a urease-positive organism, an orange or yellow is negative. The IMViC test is a series of different tests that differentiate between enterics. One is the Indole test. This test tells whether the bacterium possesses tryptophanase which is the enzyme that breaks down tryptophan into indole. The agar contains tryptic soy broth so if the bacterium contains tryptophanase, indole is produced. This production of indole is seen by adding Kovac’s reagent which causes a red ring to be seen at the top of the tube. The citrate test is also used to see which kind of products the bacteria make. It uses a green agar slant that contains sodium and ammonium phosphate. Bromythymol blue dye is late added as an indicator. Inoculation of the slant with a needle using a zig-zag then stab technique was used with the gram positive bacterium. Conversion of the medium to blue is a positive citrate result. All plates, slants, and broths were incubated at 37°C for 24‐48 hours. Results Test Unknown 16 Unknown 16 Gram Stain + - Color Yellow Yellow Shape Rod Rod Lactose n/a + Indole n/a - Urease n/a + Citrate + + Key: (+) = Positive Test (-) = Negative Test N/A = Not Used for Determination Discussion The gram

Get Access