Measuring Animal Behavior

and Evidence for Behavior

Lecture 3
 Overview
• Evaluating evidence
• Try out a real ethogram from the assigned paper
• How to design studies that ask and answer
questions about behavior
• Hypothesis testing
• Some special considerations about behavior
experiments
• Thinking about evidence
• How and why to cite your sources
 Leave an Evidence Trail

• How do you do this?

• Cite sources (using Mendeley or Zotero) to provide a
trail of evidence for others (and yourself)

• Why is this important?

• Provides support for your findings
• Provides information to your readers so they can
evaluate your evidence
 Citation Format for Assignments
• In line citations – any format that unambiguously identifies which
source in the reference section supports your assertion
• Cite your source every time you reference it in the text
• (Author et al., date published)
• Works cited section – provide the names of the authors, the year
published, the journal name, and enough information for anyone
to find the paper (e.g., title of the article, DOI, journal issue
number, etc)
• Bernaola and Holt. 2022. Incorporating Sustainable and
Technological Approaches in Pest Management of Invasive
Arthropod Species. Annals of the Entomological Society of
America 114(6): 673-685.
https://doi.org/10.1093/aesa/saab041
• Pro Tip: use Mendeley or Zotero as a free citation manager
 How to Cite Papers

• If I find a source, should I cite it, or should I cite that source’s

source?

Example:
• From Secondary Citation 2024: “Animal behavior is cool
(Primary Citation, 2020)”
• Cite the source you read
• Also as a good habit check that the primary source was
cited/interpreted correctly
• If it turns out your source is wrong, you and readers can
retrace your steps later to figure that out
 Common Citation Questions
How to cite:
• Things learned in class
• Cite the textbook or the specific lecture
referenced
• General knowledge
• Find a reliable source (from a peer
reviewed journal or government
organization) supporting the
information or re-evaluate this claim
• Individual experience
• Use words like “I observed”
• Cite your personal observation or the
observation of a knowledgeable other
(e.g., “Puppies are adorable (Holt, pers.
obs.)”)
 Common Citation Questions
How to cite:
• Sources without an author
• Avoid citing articles with no author
• Exception: information from a .gov
website (such as CDC or NIH)
• What if you can’t find a source for
something?
• Maybe that thought or belief is
incorrect
• Citations enhance the flow of writing by
supporting thoughts with evidence
 Evaluating Evidence
• Not all resources are accurate
• Remember that “general
knowledge” or “common sense”
is not a source
• Sometimes what seems
common sense or habit is
incorrect
• Not all false beliefs are “crazy”,
some are based on conditional
facts/training or misinformation
 Evaluate
Evidence Using
”Grandma’s
Ham”
• Cultural training can result
in interesting behaviors

https://medium.com/@SSDSI/grandmas-ham-a-story-of-cultural-training-e9f33413e84a
Evaluating an
Ethogram
Considerations for Behavioral Studies

Ethical considerations vary by experiment type,

animal, and country! Check these out in the papers
you find. Sometimes this section is at the end.

Who was measured

(and consider who was NOT included)

How the experiment was set up (important, but not

our focus right now)

Ideally, this section will explain how the approach

answered the questions raised in the introduction

For vertebrate subjects in a non-natural setting, an

IACUC (Institutional Animal Care and Use
Committee permit) is required!
 Evaluating Dog Behavior
• Do dogs eavesdrop on human interactions in a
helping situation?
• This paper investigates some questions about
cooperation and social information
• Basic setup:
• The dog sees one human being helpful and another
human being unhelpful to a third human (the “beggar”)
• Then the dog experiences their own “impossible task”
where help is needed to secure a reward (sausage)
• Does the dog identify which human would be more
helpful in this situation?
• Prediction: if yes, the dog will look at, approach, and interact
more with the human previously seen helping
 Ethogram for: Do dogs eavesdrop on
human interactions in a helping situation?

• Pick one of the behaviors in this ethogram

 Study Design for Observing Dog
Behavior
• Person either performs a helpful or harmful
behavior
Activity: Test the Ethogram by
Observing Dog Behavior
 Activity: Think Pair Share
Answer some questions about “Do Dogs Eavesdrop on Human Interactions
in a Helping Situation”?
• Why was the dog’s owner blindfolded?
• What behavior did you measure and what was your “answer”
(duration or frequency of the behavior)?
• Consider the reliability of person rating the behavior
• Why were the ‘helpful’ and ‘unhelpful’ humans standing an equal
distance away from the dog?
• What was your impression of Weischel the dog?
Evidence in Animal Behavior:
Understanding Types of
Studies
 Hypothesis vs. Prediction
• What is the difference between a hypothesis and a
prediction?
• Hypothesis: “An education guess” , “I think X is true”
• Prediction: “If X is true, I should observe Y”
• If X is false, I should observe something other than Y
• Helpful resource:
https://www.psychologytoday.com/us/blog/perception-
and-performance/201801/hypotheses-versus-
predictions
 Male Peacock
Spiders Mating Display

• What is the difference between a hypothesis and a prediction?

• Hypothesis: Peacock spider males gain a reproductive benefit
from their complex display
• Prediction: (If hypothesis is true) If males display, they should
have greater reproductive success than males who do not

• Null Hypothesis: If hypothesis is not true (this is the default),

males who display should have similar reproductive success
than males who do not
 Benefits and Limitations
of Scientific Studies
• Particularly with single studies
• They provide support, or fail to provide support, for
specific hypotheses
• Many hypotheses could be consistent with the
same results
• “Significant” results could be due to chance
• Maybe there are hypotheses no one has thought of
yet
 Benefits and Limitations
of Reviews and Meta-analyses
Discuss or
Test(meta-analysis)
whether multiple
independent
studies point to the
same result
 Hypothesis Testing
1. Observational study
• Observations and comparisons of different individuals (generally within a
species)

2. Manipulative study
• Manipulations of individuals or situations into two or more treatment
groups
• Treatments may mimic different natural conditions or explore behavior in
unnatural conditions

3. Phylogenetic or Comparative study

• Comparisons of the characteristics of different species (generally measured
in observational studies)

4. Theory
• Use mathematical equations, simulations, or other methods to model the
processes hypothesized
• Provides quantitative predictions and can serve as a test of whether a
hypothesized process is even possible
 Observational Study

• Observe wild animals doing behavior in an

unmanipulated state
• Sometimes with your eyes, other times with camera
traps, special recording equipment, night vision, etc.
• Observational studies can test hypotheses, e.g.,
compare observations across different individuals,
sexes, ages, seasons, times, environments, etc., or
against some null hypothesis
 Manipulative Study
• Aka: Experiment
• Typically manipulate a variable hypothesized to be related to behavior
• Compare different treatments and/or treatment vs. control
• Additional controls for aspects of the manipulation not related to the
hypothesis
• Examples
• play conspecific song vs. silence vs. random noise
• Provide animal with different food choices
• Ideally, controls eliminate other potential processes that could explain the
results
 Phylogenetic
Comparative Method
• Observe a behavior
• ask an “ultimate” question about it
• Hypothesize
• that selection in certain environments
lead to the evolution of the behavior
• Predict
• species experiencing similar
environments will also share the
behavior
• species with different environmental
factors will have different behaviors
• Test
• whether there is a correlation between
the environmental factor and the
behavior across a number of species
Example of the Phylogenetic Comparative
Method

The skulls of non-burrowing fish evolve twice as fast

as burrowing fish!
Dr. Kory Evans
 Specificity
• Interested in whether some manipulation or
stimulus influences one specific behavior (or other
process), or many different behaviors
• Examples:
• Temperature
• Predator suddenly appearing
• Caffeine – might affect more than one behavior
• Consider measuring a behavior that is the focus of
your hypothesis and some other behavior (e.g.,
general activity)
• Identifying behaviors is similar to ordering coffee
• There are many types of behaviors and coffee, so be
specific, but not overly complicated (avoid what
Leah did)
Key Differences Between Observational
and Manipulative Studies
• With observational studies you sit and watch what is
happening
• With manipulative studies variables are altered
Consequences of Differences between
Observational and Manipulative Studies

• With observational studies, it is harder to link a

specific cause (internal to the organism, or in the
environment) to a behavior
• But, with manipulative studies, you may miss a lot
of the complexities that shape the behavior in the
wild
• Potential solution: do both (for the same species
and behavior)
• Also: you can use manipulative experiment to test
some of these complexities
 Identifying Interactions
• Interaction: when the effect of something depends on
something else
• Maybe:
• Birds learn songs they hear, but only if the birds
are young?
• Dogs obey commands if they are hungry, but
ignore commands when they are not hungry?
• The sound of a predator makes younger birds less
active, but older birds more active?

• If qualify a thought (i.e., hypothesis) with:

• “but only if…”
• “but only for [a subset of the population]…”
• “actually the trend is reversed if…”
• This describes an interaction between two causal
factors
 Interaction Contributors
• Interactions can be between individuals
• Interactions can be between causes of behavior
 Types of Ecological Interactions
• Interactions can be between individuals
• Interactions can be between causes of behavior
• Can be influenced by: age, sex, genotype, microbial composition,
hormones, experiences, etc.
 Effects of Interactions
• Expect interactions between causal factors
• There are different types of interactions
 Manipulative Studies Revisited
• Vary factors in a controlled way
• Test for a causal relationship between independent (treatment)
and dependent (response) variables
• Experimental design
• Change only one factor at a time
or
• Use a factorial design to test for interactions between factors
• Perform experiments where combinations of variables
are manipulated
 Factorial Design Example
Fly covered in mold
• Some fungi are delicious and nutritious, others
are lethal
• Flies may avoid locations where they might be
infected with deadly mold
• Females feed more, so male and female
avoidance may differ

Have:
• Mold possible treatments:
• mold
• no mold
• Sex possible treatments:
• female
Fungus enters through
• male weak points in cuticle (e.g.,
segmentation, hair sockets)
 Factorial Design Example
• For a factorial design, need all possible
combinations of two or more variables

Sex
males females

yes N flies N flies

Mold N = sample size

no
N flies N flies
Flies and Fungal Spore Exposure
• Do male or female flies
avoid fungal spores (aka
conidiospores)?

Dr. C. Nick Keiser

More info: https://www.keiserlab.com/
Keiser et al 2019. Sex differences in disease avoidance
behavior vary across modes of pathogen exposure. Ethology
126:5
Flies and Fungal Corpses
• Do male or female flies
Fly corpse killed by avoid dead fly corpses
the covered in fungus?
entomopathogenic
Fly fungus
Metarhizium
robertsii

Dr. C. Nick Keiser

More info: https://www.keiserlab.com/
Keiser et al 2019. Sex differences in disease avoidance
behavior vary across modes of pathogen exposure. Ethology
126:5
Determining What Question to Ask

• The question is important and can

lead to different understandings of
organism behavior
 Harnessing Animal Behavior for
Potential Applications
• Aphids avoid leaf surfaces with bacteria Pseudomonas syringia
• The efficacy of infection may differ with bacterial strain
• This behavior can be harnessed for future potential applications of
Psuedomonas to reduce aphid pest pressures in agricultural crops

Hendry et al. 2018. Current Biology 28.; Smee et al. 2021. Appl Environ Microbiol 87(11).
 Summary

• Measuring behavior requires considerations

inherent to conducting experiments
• Plus an understanding of observer effects and
appropriate sampling techniques
• The “correct” way to do an experiment depends on
the question being asked
• Experiments do not “prove” anything, they provide
support for one or more hypotheses
 Next Class

• Hormones!

Assignments
• Reading:
• Principles of Animal Behavior (PAB) chapter 3,
• Intro and Hormone section
• Safran et al. 2008. Sexual signal exaggeration affects
physiological state in male barn swallows. Current Biology
18(11).
• Article on canvas