POLYTECHNIC UNIVERSITY OF THE PHILIPPINES

College of Science
DEPARTMENT OF BIOLOGY

BIOL 40065 MICROBIAL PHYSIOLOGY S.Y 2021-2022

NAME: ________________________________________ SCORE: _____________________

YR & SEC: ____________________________________ INSTRUCTOR: ________________

LABORATORY ACTIVITY B
Activity No. 6
CURRENT TRENDS IN MICROBIAL IDENTIFICATION

Overview

Staining, morphology, motility, culture data, enzymatic and biochemical activities can be
used to identify microorganisms. However, running a multitude of tests to identify an unknown
entail a tremendous amount of media, glassware, and preparation time that is involved just to
set up the tests. This is followed by a meticulous search in the “Encyclopedia Bergey” to
discover the identity of the specific organism. In order to reduce these factors, many rapid
multitest systems have been developed for the identification of medically important
microorganisms in a relatively short period. Moreover, miniaturized systems have the following
advantages over the macromethods: (1) minimum media preparation, (2) simplicity of
performance, (3) reliability, (4) rapid results, and (5) uniform results. These advantages have
resulted in widespread acceptance of these systems by microbiologists.

Learning Outcomes

At the end of the exercise, you should be able to:

1. Be informed on the various trends used in microbial identification.

2. Be familiarized with the advantages and disadvantages of each microbial identification
methods.

Reading Guide

• Read about the four rapid multitest systems – API 20E, API Staph, Enterotube II, and
Oxi/Ferm Tube II.
• Learn how to use the four systems and analyze their results.
• Research on how molecular techniques speed up molecular identification.

Allotment Time: Week 7 and 8 (12 hours)

Course Materials: Study the below materials taken from

https://www.academia.edu/7653687/Miniaturized_Multitest_Systems to answer the guide
questions from this activity.

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Guide Questions

RAPID MULTITEST SYSTEMS

1. For each of the following aspects, compare the four rapid multitest systems.

Oxi/Fern
API 20E Enterotube II API Staph
Tube II
System System System
System
Time Requirement
Target Microorgaism
Specimen
Preparation/ Inoculum
Requirement
Tests Utilized
Anaerobic Conditions
Interpretation of
results

2. What is the purpose of the sterile mineral oil in the API 20E System?
3. Why is the part of the wire left in the Enterotube II and Oxi/Ferm Tube II? Why is part
of the wire broken off?
4. Before using the API Staph System, what test should be performed on bacteria to
confirm that they are staphylococci? What single test differentiate
Staphylococcus aureus from other species of staphylococci?

OTHER MICROBIAL IDENTIFICATION METHOD

1. What is DNA/RNA sequencing and how is it applied in microbial identification?

2. What are the advantages of using DNA/RNA sequencing in identifying microorganisms?
3. What specific domain of microorganisms can be identified by universal PCR primers
designed to target 16s, 18s, and internal transcribed spacer (ITS)?
Domain Universal Primer Target/S
Archaea
Eukarya
Bacteria

References

Brown, A, and Smith, H (2015). Benson’s Microbiological Applications, Laboratory Manual in

General Microbiology, Short Version 13th Edition. New York, U.S.A.: McGraw-Hill Education.

Harley, J. and Prescott, L. (2002). Laboratory Exercises in Microbiology 5th Edition. New York,
U.S.A.: McGraw-Hill Education.

Madigan MT, Bender KS, Buckley DH, Sattley WM, Stahl DA. (2019). Brock Biology of
Microorganisms 15th Edition. New York, U.S.A.: Pearson-Benjamin Cummings.

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