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JOURNAL OF MICROBIOLOGY & BIOLOGY EDUCATION

DOI: https://doi.org/10.1128/jmbe.v19i2.1527

A Simple Exercise for Teaching Bacteriophage Concepts in the

Undergraduate Laboratory Using Commercially Available Disinfectant †

Latifa B. Khan1* and Hannah M. Read1,2

1Department
of Molecular Medicine and Pathology, School of Medical Sciences, The University of Auckland,
Auckland 1023, New Zealand; 2Maurice Wilkins Centre for Molecular Biodiscovery, The University of Auckland,
Auckland 1023, New Zealand

INTRODUCTION a demonstration due to the large class size and also due
to the number of other microbiology techniques taught
Bacteriophages are viruses that infect specific bacterial within the same practical class. To mimic viral plaques,
species, hijacking the infected bacterium’s machinery to we used a commercially available laboratory disinfectant,
multiply and eventually burst its prey, resulting in zones of TRIGENE (diluted at a 1:100 ratio), which is capable of
clearing on the culture plate known as plaques (1). Viruses clearing bacterial growth on the agar plate, resulting in
are difficult to visualize due to their small size, visible a visual effect similar to that of lytic phages on bacterial
only with electron microscopes (2). Many teaching facili- lawns. The teaching strategy involved analyzing phage
ties are unable to access this specialized equipment, and typing of three different culture plates (with different
other techniques must therefore be employed to explore configurations of artificial plaques, Fig. 1) of Staphylococcus
viruses in undergraduate teaching laboratories. One of the aureus, which students were told had been isolated from
most common detection methods used is a plaque assay, three hypothetical hospital wards.
whereby bacteria and bacteriophages are mixed on an agar This low-cost exercise effectively facilitated student–
plate and the plaques are observed to assess whether lytic student and student–instructor interactions, engaging
viruses specific for the bacterial hosts are present (2). The students to link theory to practical exercises in order to
plaque assay is a key laboratory method in virology for the interpret phage typing results and to discuss the answer to
isolation and enumeration of phage particles (3), as well as the question provided in the students’ laboratory manual.
for measuring virulence of the bacteriophages, and studying This created an effective collaborative learning situation in
the growth of lytic viruses (4). The assay can also be used the classroom for teaching students about bacteriophages
for the purposes of phage typing, whereby phages are used without the use of real phages.
to identify pathogenic bacteria in diagnostic laboratories. This exercise is suitable for students with undergradu-
Finally, there is renewed interest in the therapeutic use of ate experience in microbiology, virology, or associated
phages to treat antibiotic-resistant pathogenic bacterial in- laboratory techniques whose prior lectures have covered
fections (5). Teaching students about plaque assays provides introductory aspects of microbiology, viruses, and bacterio-
an understanding of the concepts of viral plaque formation phages along with safety measures taught during the course
and the host specificity of bacteriophages, comprising an and during the laboratory exercise.
important intellectual pillar in teaching microbiology.
We have designed a simple laboratory exercise in a PROCEDURE
single practical class for a second-year biomedical science
course to demonstrate the plaque assay and to use phage Materials and methods
typing to differentiate three bacterial strains of Staphylo-
coccus aureus. This phage typing exercise was designed as Groups of four students were provided with three
culture plates labeled “ward #1,” “ward #3,” and “ward
#6,” showing phage typing (Fig. 1) of S. aureus strains iso-
*Corresponding author. Mailing address: Department of Molecular lated from three different hospital wards. Preparation of
Medicine and Pathology, School of Medical Sciences, The University culture plates showing phage typing activity and detailed
of Auckland, Private Bag 92019, Auckland 1023, New Zealand. instructor notes are available in Appendix 1. Student
Phone: +64 9 3737599 ext. 84492. E-mail: [email protected]. instructions were provided in each student’s laboratory
Received: 14 November 2017, Accepted: 4 June 2018, Published: manual (Appendix 3). Clearly labeled biohazard bags were
31 August 2018. available for waste disposal, and disinfectant was available
†Supplemental materials available at http://asmscience.org/jmbe to disinfect benches.
©2018 Author(s). Published by the American Society for Microbiology. This is an Open Access article distributed under the terms of the Creative Commons Attribution-Noncommercial-NoDerivatives 4.0 International
license (https://creativecommons.org/licenses/by-nc-nd/4.0/ and https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode), which grants the public the nonexclusive right to copy, distribute, or display the published work.

Volume 19, Number 2 Journal of Microbiology & Biology Education 1

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 KHAN & READ: TEACHING BACTERIOPHAGES USING EVERYDAY MATERIALS

Phage plaques shoes, gloves, and safety glasses). After completing this
Lawn of bacteria/host cells
practical exercise, students disinfected lab-bench surfaces,
all biohazard wastes were disposed of according to the
university biohazard waste disposal guideline, and finally,
students thoroughly washed their hands in compliance with
ASM guidelines.

CONCLUSION

This phage typing exercise is easy to prepare and can

126 easily be adapted by any institution as it is or with modi-
FIGURE 1.  Student materials per group. Culture plates with simu- fication (Appendix 2) since it does not require expensive
127 FIGURE 1. Student materials pershowing
group. Culture
lated bacteriophage plaques phageplates with
typing of simulated bacteriophage
hypothetical plaques
equipment, and the materials that are needed to prepare
128 showing phage typing of hypothetical Staphylococcus aureus strains isolated from culture
Staphylococcus aureus strains isolated from three different wards: three plates to show the phage-typing activity are inex-
wards #1, #3, and #6 in the case study hospital. pensive and can easily be sourced from scientific suppliers.
129 different wards: wards #1, #3, and #6 in the case study hospital. TRIGENE disinfectant can be used as an effective tool to
Measuring students’ learning mimic bacteriophage plaques, reliably teaching students
130 about phage diversity, bacteria–virus interaction, and the
This exercise was scheduled to be completed within a process of viral plaque formation in an easily reproducible
131 15-minute period
*Corresponding author. of a two-hour
Mailing microbiology
address: Department laboratory
of Molecular manner. This quick and easy trick saves time as well as
Medicine and Pathology,
with a class size of 120 students. Students were instructed eliminating the need to source bacteriophages specific for
132 School of Medical
to record phage Sciences,
typingThe University
results of allofS.Auckland, Private Bag 92019, Auckland
aureus strains/wards different bacterial strains.
133
as outlined in Table 1 (Appendix 1). Although many plaques
1023, New Zealand. Phone: +64 9 3737599. E-mail: [email protected].
are generally round, they can be variety of shapes and sizes SUPPLEMENTAL MATERIALS
(1). A plaque is indicated by a zone of no growth (clear
134 Received: Day Month Year, Accepted: Day Month Year, Published: Day Month Year
plaque) in the lawn of S. aureus. If phages 1, 5, and 7 gave Appendix 1: Instructor’s notes
a positive result (clear plaques), the phage type would be Appendix 2: Possible modification
†
135 Supplemental
recorded asmaterials
1/5/7. available at http://asmscience.org/jmbe Appendix 3: Student’s laboratory protocol
The results were discussed by groups of students, focus-
136 ing on answering the question outlined in Table 2 (Appendix ACKNOWLEDGMENTS
1), aiming for engagement of students in active learning
137 about the relationship between bacteria and viruses, as well We would like to thank Associate Professor Simon
as to encourage teamwork and the development of critical Swift at the University of Auckland, New Zealand, for his
thinking skills. Students were also given the opportunity to assistance in designing this phage-typing exercise for teach-
question the teaching assistants if required and to learn how ing bacteria–virus interactions. We would also like to thank
to both convey and receive constructive criticism/feedback Senior Lecturer Dr. Stephen Ritchie at the University of
from their peers as well as their instructors. Auckland, New Zealand, for his valuable advice and sugges-
tions on the manuscript. The authors declare that there are
Safety issues no conflicts of interest.

Students had already received adequate training about REFERENCES

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