Experimental Studies

Ing. Pavel Jakubec, Ph.D.

Objectives
• Research
• Causality
• Problems of causality.
• How to study causality?
• Experimental design.
• Examples!
Research
• “Search for answers then search
again”
• Gather evidence and interpret it
 certain framework and
philosophies are applied
 use tested, valid and reliable
methods
 should be unbiased and
objective
Research - characteristics/requirements
• Controlled – link between cause/effect
• Rigorous – procedures relevant, appropriate and
justified
• Systematic – procedures follow logical sequence
• Valid and Verifiable – our results should be repeatable
• Empirical – conclusions based upon hard evidence
• Critical – every step of the process should be able to
withstand critical scrutiny.
Types of Research
• Application
 Pure research
 Applied research
• Objectives
 Descriptive
 Exploratory
 Correlational
 Explanatory
Causality
• Causal relationship:
• Independent
variable X (the
cause)
• Dependent variable
Y (the effect)

•X Y
[X is causing change in Y]
Causality
• Conditions:
• Association (X and Y related)
• Timer order (X precedes Y)
• Logical connection (plausible theory)
• Non-spurious relation (no competing or plausible
explanations)
Causality
• Correlation does not imply causation!!!
Ice cream example
Association
• X and Y should be somehow related!
Time order
• Cause is preceding the effect
Logical connection
• You have some plausible theory for the causal
connection.
Non-spurious relationship
• You have to rule out all competing theories that
could explaining the causal relationship. That is
the tough one.
Examples
• RQ: What is the effect of soil type on abundance
of sexton beetles?
• Independent variable?
• Dependent variable?
• Soil type Sexton beetles
Examples
• RQ: What is the effect of geomagnetic field on
behavior of dogs?
• Independent variable?
• Dependent variable?
• Geomagnetic field Dogs
Problems
• Intervening variables

• soil beetles

plants, moisture, ...

Problems
• Intervening variables (mediating variables)

• Income longevity

healthcare, food, ...

Problems
• Extraneous (Exogenous) variables

soil beetles

genetic factors, sex, ...

Problems
• Extraneous (exogenous) variables
• Causal relationship cannot be completely
described by variables under study
(endogenous variables)
• food longevity

genetic factors, sex, ...

Problems
• Spurious relationship

• ice cream # drowning

temperature (confounding
factor)
Reverse causation
•X Y
•X Y ?
• Not only the (in)famous
chicken and egg question!
Reverse causation
• Breastfeeding decrease the
growth rate of babies
• Supported by several
observational studies.
• Corrected by Marquis et al.
1997?
• Smaller babies were much
more vigorously fed despite
their genetic or other
predispositions to be small.
Example
• Q: Is a body alignment of
a dog, while he is
defecating, caused by
electromagnetic field of
Earth?
• M: Behavior was
observed and measured
in the field conditions.
• Can you prove that elm. Hart et al. Frontiers in Zoology 2013, 10:80
field is the cause?
Example
• Conditions:
• Association (X and Y related)
• Timer order (X precedes Y)
• Logical connection (plausible theory)
• Non-spurious relation (no competing or
plausible explanations)
Dogs
• Too many environmental variables are in play and there is always
something you did not measured or even think of measuring.
• Possible explanations:
• Wind direction
• Sun
.
.
.

Czech Republic Prague

Experiment
• Purpose: To investigate a causal relationship
between two (or more) variables.
• Experimental (manipulative) study (at
least two measurement moments)
• Lab experiments
• Field experiments
• Experimental (correlative) study (single
measurement moment)
How it’s done?
• Manipulation of the
independent variable by the
researcher
• Random assignment of the
research units to groups
• Control of all other variables
(source of many problems)
Conditions
• We have to be able to manipulate the
independent variable X (constrains: physics,
biology, funding,...)
• Time-lag between change of X (independent
var.) and reaction of dependent variable Y is
reasonable short (feedback).
• Ethical objections?
Unethical?
• Milgrim Study at Yale (1974)
• “Teachers” (in one room) asked
questions under the supervision of the
researcher
• “Learners” (in different room) had to
answer
• Teacher punished mistakes by electric
shocks (after each mistake they
increased the voltage)
• Only 14 out of 40 teachers stopped the
experiment before giving 450 volt shock
event though they could hear the
screams and pleads of the learner
• No one stopped before 300 volt mark.
• True purpose of the study: Test of
obedience to authority
Validity of an experiment
• Internal validity:
• Is a change of the independent variable (Y) really
caused by the change of the dependent variable
(X)?
• External validity:
• Can the conclusions of the research be
generalized outside of the experimental
situation?
Basic Experimental Design
• Pre-test (t1) to establish state before
manipulation.
t – time
Time t1 t2 O – observation
X – manipulation of
Group 1 O1 X O2 independent variable

• Was the change caused by our manipulation or

by another event?
Control group design
• Group 2 is our Control group to filter out the
influence of external events.
t – time
Time t1 t2 O – observation
Group 1 O1 X O2 X – manipulation of
independent variable
Group 2 O3 O4
• Could be the change of dependent variable
caused by systematic difference between the
groups?
Randomized control group design
• Control group and complete randomization to
filter out differences between groups.
t – time
Time t1 t2 O – observation
Group 1 R O1 X O2 X – manipulation of
independent variable
Group 2 R O3 O4 R – randomization
• What is the effect of the pre-test on value of the
dependent variable. (learning)
Solomon Four-Group Design
• True Experiment

Time t1 t2 t – time
O – observation
Group 1 R O1 X O2 X – manipulation of
Group 2 R O3 O4 independent variable
R – randomization
Group 3 R X O5
Group 4 R O6
• We are able to identify or estimate most threats
to internal validity of the experiment.
Experimental artifacts
• Experimenter effect (wishful
thinking) – Different
interaction with experimental
and control group (one gets
extra attention)
• Placebo effect – Effect of
belief that participant is treated
• Contagion – Effect of
experimental and control group
communication

• Solution: Blind/ double blind

Problems of experiments
• Complete randomization
is not always the best
choice (impractical)
• Sample size matters
• Your experiment will be
most likely called artificial
More examples
• Eggs (and other instars) of invasive
ladybird H. axyridis contain
parasitic microsporidia.
• Interspecies predation and
cannibalism is common among
ladybirds.
• Injection of these microsporidia
resulted in higher mortality (100%)
(Vilcinskas et al. 2013).
More examples
• Q: What is the effect of ingestion of H. axyridis’s
eggs on the mortality of seven-spot ladybird (C.
septempunctata)?
• Independent variable?
• Dependent variable?
• Harmonia’s eggs mortality
More examples
• Experimental design

Time t1 t2 .... tn
Group 1 R O1 X O2 Oi
Group 2 R O3 O4 Oj
More examples
Vilcinskas et al.
(2013)
More examples
More examples
• Conclusion: Adults of C.
septempunctata are not
suffering with higher mortality
after consummation of (5) eggs
of H. axyridis.
• What is the cause of resistance?
=> another story!
• Was the dosage high enough?
More examples
• Q: Is smoking
responsible for higher
occurrence of lung
cancer?
• How to design
experiment, which
could answer this
question?
Summary
• Research\Science as way of thinking
• Causality
• Problems of causality (intervening and
external variables + reverse causation).
• Experiments
• Experimental design (randomized
control group design).
 Take home message

Correlation does not imply causation!

AND
Question everything!
References:
• Kumar, Ranjit (2011): Research Methodology (a step-by-step guide
for beginners). 3rd edition, Sage Publications Ltd, Los Angeles.
• Ruxton, Greame D., Colegrave, Nick (2010): Experimental Design
for the Life Sciences. 3rd edition. Oxford University Press, Oxford.
• Vilcinskas, A., Stoecker, K., Schmidtberg, H., Röhrich, C.R. & Vogel,
H. (2013) Invasive harlequin ladybird carries biological weapons
against native competitors. Science (New York, N.Y.), 340, 862–3.
• Track your dog defecation:
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• https://play.google.com/store/apps/details?id=ian.com.essen&hl=c
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