Title: Write your title here
A Protocol for Assaying Swarming Motility in Microbes
Yangtsho Gyaltshen1 and Brooke Jude1
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1
Bard College: Biology Program
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Type of Manuscript: CourseSource Teaching Tools and Strategies Manuscript
Funding & Conflict of Interest Statement:
Bard College: Biology Program
List of Tables, Figures and Supplemental Material:
Figure 1. Model of swarming assay protocol and swarming motility over time
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Abstract Page
The abstract should be a single paragraph of 250 words or less. Start with an opening sentence
that sets the teaching challenge that you address in this manuscript, provide background
information and briefly describe your take-home message.
This swarming motility assay protocol tool is aimed at enhancing a visual understanding of
morphological variability among bacterial strains of environmental isolates. The assay is a
variation on traditional Swarming motility protocols modified by a student to characterize motility
differences in the transposon mutant strains of iodabacter sp. This serves as a beneficial tool as
it can be harnessed as a visual model for genetic for analysis of a gene and subsequent protein
function. The simplicity and low-costs associated with this technique make its implementation in
variable biology classrooms easily applicable. In addition to simplicity, various media bases can
be utilized by reduction of agar concentrations to optimize growth conditions for the microbe of
interest. This visual nature of this tool encourages students to apply genetic characterization of
microbial to phenotypic variability under different growth conditions.
Teaching Tools and Strategies Context Page: To make the submission process easier, you
may want to fill out the following form, (you will be asked to select answers during the
submission process). Choose all applicable options that effectively describe the Teaching Tools
and Strategies.
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submitting on the website.
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Course
Biochemistry
Cell Biology
Genetics
Microbiology
Molecular Biology
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Course Level
Upper Level
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Class Type
Lecture
Lab
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Audience
Life Sciences Major
4-year College
University
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Class Size
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1 – 50
51 – 100
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Lesson Length
Multiple class periods
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Key Scientific Process Skills
Reading research papers
Reviewing prior research
Asking a question
Formulating hypotheses
Designing/conducting experiments
Predicting outcomes
Gathering data/making observations
Analyzing data
Interpreting results/data
Communicating results
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Pedagogical Approaches
Think-Pair-Share
Brainstorming
Case Study
Collaborative Work
One Minute Paper
Reflective Writing
Concept Maps
Strip Sequence
Computer Model
Physical Model
Interactive Lecture
Pre/Post Questions
Other
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Bloom’s Cognitive Level (based on learning objectives & assessments)
Foundational: factual knowledge & comprehension
Application & Analysis
Synthesis/Evaluation/Creation
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Principles of how people learn
Motivates student to learn material
Focuses student on the material to be learned
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Develops supportive community of learners
Reveals prior knowledge
Requires student to do the bulk of the work
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Vision and Change Core Concepts
Structure and Function
Information flow, exchange and storage
Pathways and transformations of energy and matter
Systems
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Vision and Change Core Competencies
Ability to apply the process of science
Ability to use quantitative reasoning
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Key Concepts: List 3 – 10 key concepts (topics) that are relevant for the Teaching Tools and
Strategies (e.g. plating; PowerPoint; Large Class; Writing; etc.)
o Plating
o Pipetting
o Basic photography
Main Text
Begin the Teaching Tools and Strategies text on a new page.
1. Main Text: Describe how to use the resource, tool or approach in as brief and practical way
as possible, citing references and related materials.
This tool was introduced as a method to assay morphological differences in Iodobacter sp.
mutants created in a laboratory setting (Micro module). This is a qualitative visual method to
characterize this environmental isolate in respect to its role in bioaugmentation against the
fungal pathogen, Batrachochytrium dendrobatidis.
This assay was utilized to observe morphological differences in transposon mutants of the
Iodobacter sp. This method allowed us to characterize the mutants and can be harnessed as a
qualitative tool in genetic analysis of a gene and subsequent protein function.
Swarming motility is an organized surface translocation has been observed in both Gramnegative and Gram-positive species and requires the sensing and integration of a variety of
environmental, as well as intracellular, signals involving surface contact and local high
population density. This serves as an important tool in understanding both cellular motility as
well as cell-to-cell interactions.
This simple method utilizes a basic understanding of plating and pipetting techniques and can
be implemented with a variety of media bases. All materials used by this tool are available in a
college laboratory in addition to basic photographic hardware for imaging purposes. Overnight
cultures of your organisms of interest are prepared in a liquid media of choice. They could
include, but are not limited to 1% tryptone, R2A, and LB liquid media. Your isolates are
inoculated into the liquid media and placed on a rotator overnight at room temperature (for
iodobacter sp.). The following day, you would carefully pipette 3 uL of the culture onto the center
of your swarming plates (eg. 1% Tryptone, R2A and LB) consisting of 0.5% agar (5g/L),
allowing the drop to air dry before turning over the plates and storing them at optimum growth
temperature. A caveat to keep in mind is the possibility of plate contamination during air drying.
For the best results, air drying should be done under a sterile hood and the lids should be
replaced as soon as possible. Multiple isolates can be observed on the same plate to observe
swarming interactions. These inverted plates are incubated at the organism's ideal temperature
for 24 to 48 hours before making swarming observations. Swarming motility and growth patterns
are observed and photographed using a flatbed scanner daily over the course of ten to fourteen
days (depending on speed of growth)
The laboratory techniques used can be implemented at even an introductory course level to
make inferences about morphological differences between different organisms. However, the
genetic mechanisms underlying these interactions are better suited for upper level biology
students.
2. Scientific Teaching Themes: Explain how the Teaching Tools and Strategies relate to the
Scientific Teaching Themes of:
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Active Learning: How will students actively engage in learning the concepts? List and/or explain
the active learning strategies that are used in the tool. For example, activities could include think-pairshare, clicker questions, group discussion, debate, etc. Include both in-class and out-of-class activities.
The practical and visual implementation of key concepts serves as a pleasant method to help
students grasp morphological differences in bacterial species of their choice. Integrating a
qualitative visual category to the course will allow students to make assessments of phenotypes
that were previously uncategorized.
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Assessment: How will teachers measure learning? How will students self-evaluate their learning?
List and/or explain the kinds of assessment tools used to measure how well students achieved the learning
objectives. For example, assessments might be clicker questions, forced choice questions, exams, posters,
etc.
This laboratory technique is a good way to assess practical reasoning and hypothesis
construction in biology students. Lab reports of their observations would be assessed through
their construction of logical interpretations of results as well as their capacity to grow plates that
were relatively contaminant free. The replication of these results would be an important factor to
assess within their given framework of funds.
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Inclusive Teaching: How is the Teaching Tool and Strategy designed to include all participants
and acknowledge the value of diversity in science? List and/or explain how the Teaching Tool and
Strategy is inclusive and how it leverages diversity in the classroom and beyond. For example, the
Teaching Tool and Strategy may use multiple senses and provide examples of scientists from different
backgrounds.
Visual strategies enhance discussion within the classroom and between peers outside of regular
class hours. Due to the simplicity of the skills required to set up a swarming plate, it can be
incorporated into classes with varying degrees of complexity. The total time involved in setting
up these plates is also minimal, with only short bursts of time allocated to image the changes in
growth. The visual appeal of the patterns that emerge from swarming draws students who may
not normally be as inclined toward laboratory work. This would encourage sharing of results and
peer reviews of observations.
v Subheadings: can be included within the sections above to increase readability and clarity.
Acknowledgments
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paragraphs.
Dr. Brooke Jude
References
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* Cite references in the text using superscript Arabic numbers. Use commas to separate
multiple citation numbers. Superscript numbers are placed outside periods and commas and
inside colons and semicolons.
1. Begin the reference list on a new page. The reference list is comprehensive and spans the
text, figure captions and materials.
2. Number references in the order in which they appear in the text. Follow ASM style and
abbreviate names of journals according to the journals list in NCBI. List all authors and/or
editors up to 6; if more than 6, list the first 3 followed by “et al.” Note: Journal references should
include the issue number in parentheses after the volume number.
Examples of reference style:
1. Knight JK, Wood WB. 2005. Teaching more by lecturing less. Cell Biol Educ. 4(4):298-310.
2. Samford University. How to get the most out of studying: A video series.
www.samford.edu/how-to-study/. Accessed August 20, 2013.
3. Handelsman J, Miller S, Pfund C. 2006. Scientific Teaching. New York, NY:W.H. Freeman.
3. Please add notes to the end of the reference list; do not mix in references with explanatory
notes.
Figure and Table Legends
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* The actual figures, tables, and supplemental materials are uploaded as separate
documents and should not be included in this text file.
Figures:
Figure. A sample model of techniques used during for the Swarming Motility Assay and the
same plates with iodabacter sp. growth after ~72 hours and after 10 days of growth at room
temperature.
Supplementary Materials:
Swarming Agar composition:
R2A Swarming media:
The R2A swarming media utilizes 0.5% agar (5g/L), Proteose peptone (0.5g/L) , Casamino
acids (0.5g/L), Yeast extract (0.5g/L), Dextrose (0.5g/L), Soluble starch (0.5g/L), Dipotassium
phosphate (0.3g/L), Magnesium sulfate 7H2O (0.05g/L), Sodium pyruvate (0.3g/L) at a final pH
of 7 ± 0.2 at 25 °C
LB swarming media:
The LB swarming media consists of 0.5% agar (5g/L), Tryptone (10g/L), NaCl (5g/L), and yeast
extract (5g/L)
1% Tryptone swarming media:
The tryptone swarming media consists of 0.5% agar (5g/L) and tryptone (10g/L)