Microbiology Laboratory Report
Date: 18/02/2025
Name: [Your Name]
Department: Microbiology
Institution: [Your Institution]
1. Water Analysis
Water analysis is the process of testing water quality to determine its safety for drinking
and other uses. The pH of clean water should be between 6.5 and 8.5. If it is too low or too
high, the water is unsafe for consumption.
To check for bacterial contamination, we collect a water sample and introduce it to a
nutrient medium such as MacConkey agar or Eosin Methylene Blue (EMB) agar. After 24
hours of incubation at 37°C, we examine the plates for bacterial growth. If bacteria grow,
the water is considered contaminated.
2. Water Inoculation
Water inoculation involves introducing a water sample into a sterile medium to check for
microbial contamination. The steps include:
1. Measuring 25g of agar and dissolving it in 1000ml of distilled water.
2. Pipetting 1ml of the water sample and introducing it into a sterilized Petri dish.
3. Using the pour plate method, where nutrient agar is poured over the inoculated sample.
4. Incubating the plates for 24-48 hours at 37°C to allow bacterial growth.
5. Observing colony formation to determine contamination levels.
3. Serial Dilution
Serial dilution is a technique used to reduce the concentration of bacteria in a sample. It
helps in obtaining countable colonies for microbial analysis.
Steps:
1. Take 9ml of sterile distilled water in a test tube.
2. Add 1ml of the bacteria sample to the test tube and mix well. This is the first dilution
(10⁻¹).
3. Take 1ml from this tube and transfer it into another tube with 9ml of sterile water. This
creates the second dilution (10⁻²).
4. Repeat the process to achieve further dilutions (e.g., 10⁻³, 10⁻⁴, etc.).
5. Pipette 1ml from the last test tube onto an agar plate and incubate at 37°C.
6. After incubation, count the colonies and multiply by the dilution factor to estimate the
bacterial count in the original sample.
4. Gram Staining
Gram staining is a laboratory technique used to classify bacteria as Gram-positive or Gram-
negative based on cell wall composition.
Steps:
1. Prepare a bacterial smear on a glass slide and heat-fix it.
2. Apply crystal violet stain and leave for 60 seconds.
3. Rinse and add iodine solution for 60 seconds to fix the stain.
4. Decolorize with alcohol for 30 seconds.
5. Counterstain with safranin for 60 seconds.
6. Rinse and dry the slide.
7. Observe under a microscope using oil immersion (100x objective lens).
Gram-positive bacteria appear purple, while Gram-negative bacteria appear red or pink.
5. Use of the Autoclave
An autoclave is a device used for sterilizing laboratory instruments, culture media, and
biological waste using high-pressure steam.
Operating the autoclave:
1. Load the materials into the autoclave chamber.
2. Set the temperature to 121°C and pressure to 15 psi.
3. Run the sterilization cycle for 15–20 minutes.
4. Allow the pressure to release before opening the chamber.
5. Remove sterilized materials using heat-resistant gloves.
