Name: Buenaflor, Andrea Mae P.

Date Submitted:
Enorio, Isabelle D.
Reyes, Shylene Kaye A.
Rivera, NJ Aleah F.
Yap, Fhardina D.
Date performed:

I. Introduction

A procedure or treatment known as decontamination makes a tool, instrument, or

work surface safe to handle. Decontamination techniques can range from straightforward
cleaning with soap and water to sterilizing using an autoclave or ethylene oxide.
Decontamination techniques include antisepsis, disinfection, and sterilization. Sterilization
is the process of eliminating all microbes, including bacterial endospores that are
incredibly resistant, through physical or chemical means.

Decontamination is a vital component of microbiological safety practice and serves

to protect students from contamination, potential infection, and the release of infectious
organisms to the outside environment. Decontamination of media, work surfaces, and
equipment is also necessary to prevent contamination of cultured organisms.

II. Objectives
● To make essential steps of an effective decontamination protocol
● to acquire the ability to decontaminate a contaminated medium.
III. Requirements:

A. Materials: Erlenmeyer flask, petri dish, hot plate, culture test tubes, beaker, 10mL
syringe, alcohol lamp, appropriate personal protective clothing, antimicrobial liquid
soap, wire brushes, trash container, paper towels, galvanized tubs, tap water, and
distilled/deionized water, distilled water
B. Chemicals: Nutrient broth, and nutrient agar
IV. Procedure

Decontamination

Infectious wastes such as contaminated liquids and solids will be handled, treated,
and disposed of according to biological waste policies and procedures. Liquid wastes such
as bacterial or viral culture media will be treated with appropriate disinfectants prior to sink
disposal. Solid wastes from the contaminated media will be segregated and buried in a
closed area where they cannot get in contact with them.

● Put on proper PPE such as lab gown, gloves, head cap, face mask
● Place the small basin beside the sink for the solid and liquid waste from the
contaminated media.
● Solid wastes from the contaminated media will be segregated and buried in a closed
area.
● Place an absorbent material (paper towel, rack) over the uncontaminated surface,
wash the petri dishes thoroughly in a running water with an antimicrobial soap.
● After washing, rinse thoroughly and place it in an absorbent paper towel.
● Allow sufficient contact time after applying the disinfectant.
● Rinse the cleaned area with distilled water to avoid adverse effects on the
experiment.
Sterilization

When microscopic biological media have been made, they still have to be sterilized
because of microbial contamination in the air, on glasses, and on hands. Within a few
hours, there will be thousands of bacteria reproducing in the medium, so it has to be
sterilized quickly before the microbes start using the nutrients.The sterilization process is
100% effective and guarantees that the medium will stay sterile. Media sterilization is
carried out in the autoclave. When the pressure from the steam is at a certain point in the
jacket, a valve allows the steam to enter.

The usual procedure for using the autoclave to give the medium to the biochemistry
lab because the personnel in charge will be the one autoclaving
 V. Conclusion

We concluded that there are essential things that we should take note of during the
dispensing of media. In dispensing the agar, it must be done close to the alcohol lamp to
maintain the sterility of the procedure. Then always remember to pass lightly the materials
in the alcohol lamp before and after opening. Also, when pouring tha agar to the petri dish,
time should be observed as the agar is solidifying. This procedure was repeated due to the
agar being contaminated. On the other hand, in dispensing the broth, accuracy of
measurements and sterility must be noted. It has been realized that the procedures must be
properly done in order to produce the expected results. Each measurement of the chemical
and material are significant in obtaining the objectives of this laboratory activity, which are
mainly to introduce both nutritional agar and broth and to isolate a pure culture.
VI. Documentation
a. Decontamination
b. Sterilization
VII. References

● Mettler-Toledo International Inc. all rights reserved. (2022, November 16). Culture
Media Preparation. Mettler-Toledo International Inc. All Rights Reserved.
https://www.mt.com/ph/en/home/applications/Laboratory_weighing/Culture-Media-Prep
aration.html
● Aryal, S. (2022, August 10). Nutrient Agar: Composition, Preparation and Uses.
Microbiology Info.com.
https://microbiologyinfo.com/nutrient-agar-composition-preparation-and-uses/#:~:text=
Water%20is%20essential%20for%20the,various%20nutrients%20can%20be%20transpo
rted.
● Laboratory Exercises in Microbiology: Discovering the Unseen World Through
Hands-On Investigation Petersen & Susan McLaughlin
https://academicworks.cuny.edu/cgi/viewcontent.cgi?article=1015&context=qb_oers