Experiment No: 02

Experiment Name: Sampling of Microbs from a Spoiled Food.

Introduction
Spoilage of food is a natural process caused by the growth of micro-
organisms such as bacteria, fungi, and yeasts. These micro-organisms
utilize nutrients present in the food for their own growth and
metabolism, leading to changes in the food's appearance, odor,
texture, and taste. The specific type of spoilage and the micro-
organisms involved depend on the type of food, storage conditions,
and initial microbial contamination.
Objective
The main objective of this lab report is to describes the procedures for
sampling micro-organisms from spoiled food and the subsequent
analysis to identify the spoilage-causing microbes.
Requirement
i. Sterile swabs or spatulas
ii. Sterile sample containers
iii. Enrichment media
iv. Selective agar plates
v. Gram stain reagents
vi. Microscope
vii. Incubator
viii. Spoiled food (Banana)
Procedure
1. Visual inspection of the spoiled banana was performed to
identify areas of discoloration, slimy texture, or mold growth Sterile
swabs or spatulas were used to collect samples from these areas.
Samples were placed in sterile containers and kept refrigerated until
further processing.
2. Depending on the suspected type of spoilage, appropriate
enrichment media were used to promote the growth of specific
microbial groups. Samples were inoculated into the enrichment media
and incubated at the appropriate temperature for a specific time.
 3. After enrichment, a loopful of the enriched culture was
streaked onto selective agar plates. These plates contained nutrients
and chemicals that favor the growth of specific groups of
microorganisms while inhibiting others. Plates were incubated at the
appropriate temperature for a specific time.
4. Colonies appearing on the agar plates were examined for
their size, shape, color, and other characteristics. Gram staining was
performed to determine whether the bacteria were gram-positive or
gram-negative. Additional biochemical tests may be needed for
further identification.

Result
The microscopic examination of the Gram-stained slides revealed the
presence of micro-organism in the spoiled food. Colonies on the agar
plates were seen clearly also. Based on the microscopic and cultural
characteristics, the spoilage-causing microorganisms were identified
as Escherichia sp. and Klebsiella sp. from spoiled banana.
Discussion
The results of this study indicate that the spoilage of the banana was
caused by Escherichia sp. and Klebsiella sp.
The identification of the spoilage-causing microorganisms is
important for several reasons:
i. It helps to determine the appropriate storage conditions and
handling practices to prevent future spoilage.
 ii. It allows the development of effective food preservation
methods that target specific microbial groups.
iii. It provides information about the potential risks of foodborne
illness associated with the consumption of spoiled food.
Precautions
i. Sterilize sampling tools to prevent cross-contamination between
samples.
ii. Collect samples from multiple locations within the spoiled food.
iii. Work quickly to minimize exposure of the sample to the
surrounding environment.