Microbiology Laboratory Report (8)
Name - Sahra Abdulkadir ID - MD/DGR/0022/22 Date - 05/08/2024
Bacterial Identification
Introduction
Bacterial identification is the technique used to identify the particular species or strain of bacteria that
is present in a sample. We can identify bacteria using many different methods such as microscopic
examination, cultural appearance, biochemical reaction, serological identification, animal inoculation,
bacteriophage typing and molecular method (genomic test). The aim of identifying bacteria is to
classify bacterias at the species level and further make proper and accurate clinical decisions such as
diagnosis and treatment planning.
Objective
Learn the cultural characteristics of different kinds of bacterias i.e their shape, size, pigment,
hemolytic behavior, fermentation etc. Practice some of the biochemical tests used to identify
bacterias.
Materials Used
- Gloves - Bacterial culture media (blood agar)
- Hydrogen Peroxide solution - Pipette
- Glass slide - Blood sample (plasma)
- Inoculating loop - Bunsen burner
Procedure steps
A. Catalase test
1. Put on gloves. Prepare a glass slide. Add a few drops of hydrogen peroxide on the glass slide.
2. Sterilize inoculating loop. Open bacterial culture media and take some bacteria. Add onto slide.
3. Slowly mix the bacteria with the hydrogen peroxide and evenly distribute it around the slide.
4. Observe for bubble formation. Sterilize inoculating loop.
B. Coagulase test
[Link] a glass slide. Open blood sample, take some using pipette. Add onto [Link] pipette.
2. Sterilize inoculating loop. Open bacterial culture media, take some bacteria. Inoculate onto slide.
3. Slowly mix the blood(plasma) with the bacteria while evenly distributing it around the slide.
4. Gently swirl it around the slide. Observe for any agglutination reaction
Result
A. Catalase test - a lot of bubble formation was seen, indicating a positive catalase test.
B. Coagulase test - clot formation was seen, indicating a positive coagulase test.
Interpretation
Catalase test is a biochemical test used to differentiate between Staphylococcus and Streptococcus.
The genus Staphylococcus has catalase enzyme, which converts hydrogen peroxide into water and
oxygen. During a catalase test, these bacterias present with bubble formation because of the oxygen.
Hence they are catalase positive. On the other hand, the genus Streptococcus doesn't have catalase
enzyme so no bubble formation is seen during catalase test, therefore they are catalase negative.
Coagulase test is a biochemical test used to further classify the Staphylococcus bacteria on the species
level after we confirmed a positive catalase test. Coagulase is an enzyme that converts fibrinogen into
fibrin facilitating clot formation. A positive coagulase test indicates the presence of clot formation
therefore confirming the bacteria as Staphylococcus aureus.
Result (cultural characteristic of previously cultured bacterias) - Group 1 *Pleural fluid*
- Nutrient agar - greenish discoloration that is of diffuse type (exopigment)
- MacConkey agar - Pink color seen therefore indicating presence of lactose fermenter.
- Blood agar - complete hemolysis is seen indicating presence of beta hemolytic bacteria.