Republic of the Philippines

University of Sto. Tomas

College of Science

Exercise 6 in Microbiology 200L:

Isolation of Microorganisms from Environmental Samples

Submitted to:

Ms. Anna Christina Ngo, M.Sc.

Asst. Prof. Michael C. Valdez

Submitted by:

Santos, Shena Kyle

Sta. Maria, Diane Samantha

Tusing, Tyra Isabelle

Uy, John Kenneth

(GROUP 9-2BIO7)
I. Introduction

In our everyday experiences, microbes are virtually ubiquitous. The natural

environment is populated with many organisms suspended in the air we breathe, the

food we eat, the water we drink, and they are also associated with other organisms (e.g.,

humans). Microorganisms contaminate the surfaces of objects in the laboratory and in

the environment which can be a potential source of contamination. Armed with this

information, sterile techniques have been developed to minimize laboratory

contamination. To contaminate, is to accidently grow an unwanted organism. The

identification of bacteria is a careful and systematic process that uses many different

techniques to narrow down the types of bacteria that are present in an unknown

bacterial culture. It produces benefits for many aspects of the research of

microorganisms and helps physicians correctly treat patients.

II. Objectives

1. To enumerate the different microbial species present in a given environment.

2. To isolate microorganisms from environmental sources.

III. Materials

Nutrient Agar Plates Distilled Water

Cotton Swab (sterile)

Potato Dextrose Agar Plates

IV. Procedure

A. Interrupted Multiple Streaking

inoculum was dragged to make the primary streak on the first quadrant,

occupying about half the plate. The inoculum was flame-sterilized. The plate was

turned approximately 90o. The inoculum was touched to the part of the first

quadrant then was dragged on the second quadrant to make the second

streaking. The same procedure was repeated until the fourth quadrant for the

fourth streaking.

B. Airborne Microorganisms

1/2 strength Nutrient Agar(NA) and Potato Dextrose Agar(PDA) plates were

prepared. A NA and a PDA plate was exposed in the Benavides Library for 1-2

minutes. The plates were incubated at 35oC for 24 hours. Different colony forms

were observed on the culture media.

C. Microorganism on Banana Peel

A sterile cotton swab and a sterile 3-ml H2O was prepared. The peel of

banana was swabbed with the sterilized cotton swab moistened with the

sterilized H2O. The sterilized cotton swab was spread on the surface of the 1/2

strength NA medium and PDA medium. It was incubated at 37oC for 24 hours.

Microbial growths were observed.

D. Microorganisms in Human Skin

A sterile cotton swab and a sterile 3-ml H2O was prepared. The chin was

swabbed with the sterilized cotton swab moistened with the sterilized H2O. The

sterilized cotton swab was spread on the surface of the 1/2 strength NA medium

and PDA medium. It was incubated at 37oC for 24 hours. Microbial growths

were observed.

V. Data and Results

Applying Wet Cotton Swab

PDA and NA Plate Library

PDA and NA Plate Banana

proper PH, proper temperature, and gasses. The survival of different microorganisms in the

environment depends on the different conditions provided by the environment. There are some

that can grow on extreme temperature and salinity. Some can even grow without oxygen. Microbial

growth means the growth of different microbes in number and not in size which indicates their

growth through reproduction.

In this experiment different cultures were cultured from different environmental samples,

PDA and half strength NA plates were prepared for the samples while observing aseptic technique.

Half strength NA plates contain only half of the nutrients originally provided by the culture medium

but having the same percentage of agar for the culture medium to solidify. Half strength of NA was

used so that microorganisms will not crowd the plate and so that only bacteria that can benefit

from the nutrient of the culture medium will grow. Both PDA and NA plates are able to culture

microorganisms, however their difference from each other is that NA plates are more suitable for

the growth of fungi bacteria while PDA plates are more suitable for the growth of fungi because of

the potato infusion component present in it.

In the library culture sample, microorganisms grew only on the NA plate. Because the plates

were only exposed to air and did not touch any solid surfaces, the transfer of microorganisms to

the culture was at a very slow rate. And also, the plates were only exposed on air for a minute which

limited the microorganisms that can transfer to the plate. In the human skin culture sample, it is

important that the distilled water and the cotton swab used were sterile to avoid contamination. It

is important that he cotton swab used must be wet before applying it to the skin because the

bacteria will adhere more on the cotton swab. In the 24-hour culture plates from human skin,

microorganisms grew on both plates which indicate that fungi and bacteria are both present on the
human skin. However, since there are more colonies visible/ present on the NA plate it can be

concluded that there are more bacteria present on the human skin sample than fungi. There are no

microbial growth present/visible in the PDA plate culture sample of the banana peel while there

are visible microbial colonies on the NA plate which indicates that bacteria are present on the ripe

banana however, it can also be concluded that since the banana have been on many surfaces

already, and was touched by different people, the bacteria on the banana peel were caused by such.

According to a research, the cause of the ripening of the banana is because of the ethylene that can

be produced by plants and some bacteria. This microorganism may be present in the banana

culture sample aside from the contaminants. Microbes from the ice cream sample both grew on the

PDA and NA plates which indicate that there can be some fungi and bacteria in the ice cream

sample.

VI. Conclusion

Different microbes were isolated from the different environmental samples, but the

species were not yet identified. Most cultures from the different environmental samples

contain more bacteria than fungi.

VII. References

Atlas, R. (2010). Handbook of microbial media (4th ed.). CRC Press

Bahrami-Hessari, M., Dedeles, G. R., De Jesus, M. M., Dela Cruz, T., Papa, D. D., Quinto, E.

R., & Santos, M. G. (2014). Laboratory manual in general microbiology (8th ed.).

Department of Biological Sciences.

V, Avi D, Ferrentino G, Dorigato A, Torre P, Jousson O, Mansy SS, Del Bianco C. (2014)

Ethylene-producing bacteria that ripen fruit. :935-8. doi: 10.1021/sb5000077

Oyewole, O.A. (2012). Microorganisms Associated with Deterioration of Stored Banana

Wilson C. (2007). Microbial food contamination. CRC Press

Wet is better for bacteria; produce cross-contamination in the kitchen Retrieved on

March 18, 2016 http://barfblog.com/2013/09/wet-is-better-for-bacteria-produce-cross-

contamination-in-the-kitchen/