SCIENTIFIC METHOD

SPECIFIC OBJECTIVES
1. Define the following terms: observation, hypothesis, prediction, test, theory, postulate, law, falsify, support.
2. Understand the reasoning process known as hypothetico-deductive scientific inquiry
3. Identify the subjective and objective parts of hypothetico-deductive inquiry.
4. Be able to explain how a hypothesis is tested and what the possible results are.
5. Explain why a hypothesis can be falsified but not proven.
6. Understand the value of falsifying a hypothesis.
7. Know the characteristics of a good hypothesis, a good prediction, and a good test.

BACKGROUND
"Science" is a search for understanding. Science is much more than the simple collection of facts. The hypothetico-
deductive approach to science is a reasoning process that challenges us to consider our observations, propose a
unifying explanation for these observations (a hypothesis), and then to test our putative explanation. On the basis of
such tests, the hypothesis may be falsified (shown to be untrue), or it may be supported. If the hypothesis is falsified,
then we must return to the observations and draw on our imagination and insight to devise another hypothesis. If the
hypothesis is supported, then we devise other ways to test the hypothesis. If a hypothesis survives repeated rigorous
testing, and is sufficiently general, it may come to be regarded as a theory. More commonly, theories are logical
constructs that meld several related hypotheses in a way that permits a family of predictions and explanations that
are broader than could be derived from any component hypothesis. In this context, hypotheses function as postulates.
Historically, some theories have become so highly regarded that have become expressed as laws (but this is usually
an artificial distinction and the term “law” will probably not be applied much in the future). Theories are the ultimate
product of science. Theories provide a framework of understanding about the universe, and serve as the basis for
new observations, new questions, and new hypotheses. Mature sciences are characterized by a body of general, well
substantiated, theories. Sciences grow through the addition, refinement, and rejection of theories.
It is fundamental to hypothetico-deductive science that hypotheses can be falsified, or convincingly supported, but
never proven. This is not to say that we know nothing with certainty, but rather to recognize to there is a spectrum of
certainty. The two laws of thermodynamics are examples of a theory in which we have great confidence. Predictions
of these “laws” have been shown to be true in test after test after test. At least for now, the laws of thermodynamics
seems so likely to be true that to deny them would seem absurd. “Fick’s Law of Diffusion” is an example of a
physiological theory that has survived considerable testing and in which we have great confidence. The optimum
allocation hypothesis as articulated by Bloom et al. 1985 to explain patterns in plant resource allocation is an
appealing but far less mature hypothesis. I am reasonably certain that important components of the optimal
allocation hypothesis are true, yet I would be surprised if it survives the next 10 years entirely intact.

"Science is the art of the soluble " - P. Medawar

The chief difference between science and nonscience is testability. Any
hypothesis could be interesting, and any explanation could be true, but a
hypothesis does not qualify as science unless it is testable. The
hypothesis "there is a God" is certainly interesting and could certainly be
true, but is nontestable. W e can devise no experiment where one possible
result would support the hypothesis and another would falsify the
hypothesis. Therefore, the existence of God is not a problem that can be
solved using the scientific method. It is an issue of philosophy, not
science. The world is full of hypotheses that are interesting but
nontestable; all such nontestable propositions are beyond the bounds of
science. The job of a scientist is to devise hypotheses that are both
interesting and testable.

Fig. 1. A recipe for scientific progress

is to tackle questions that are
challenging but tractable.
 T HE SCIENTIFIC METHOD ILLUSTRATED WITH AN EXAMPLE

Fig. 2 summarizes the scientific method in schematic form. Refer to this as you consider the following example
drawn from the work of Dr. Erkki Haukioja and his colleagues at the University of Turku in Finland. This research
elegantly illustrates the principles of good science and has been exceptionally influential in shaping our ideas about
plant-herbivore interactions.

Fig. 2. Schematic of scientific method.

OBSERVATIONS
Much of the northern Scandinavia is dominated by forests of mountain birch (Betula pubescens ), a species that is
related to the white birches of North America. In Scandinavia, these birch forests are subject to occasional
spectacular attacks by the larval stage (caterpillar, inchworm) of a particular moth (Epirrita autumnata;
Geometridae). These insect outbreaks, or population explosions, commonly cause the defoliation of thousands of
acres of birch forests, and sometimes kill considerable numbers of trees. If you have ever seen outbreaks of gypsy
moths, forest tent caterpillars, spruce budworm, bark beetles, or other insects, you have some appreciation for how
spectacular such an outbreak can be. In Scandinavia, where good records exist, large outbreaks have occurred about
12 times in the last 120 years, and it seems likely that such outbreaks have been occurring throughout the Holocene.

QUESTIONS
W hy do these insect populations fluctuate so dramatically? In particular, what causes the abrupt population decline
(crash) that nearly always terminates the outbreak? W hy do the insects not remain abundant until the forest has been
killed and no food remains? W hy are trees not killed more often than they are by these outbreaks? Predators and
diseases provide the most common answer to these questions, but Haukoja advanced an explanation that does not
require predators or diseases.

HYPOTHESIS
Birch trees have an inducible defense system to protect themselves from repeated insect attacks. ("Inducible" implies
that the trees are not always well defended. Rather, the defensive response is triggered by some environmental cue,
such as insect grazing. W hat would be the advantage of an inducible defense system versus one that was always in
effect?)
PREDICTION
Birch trees that have been damaged by insects in one season will, in the next season, produce leaves that are of low
food quality for the insects. Insects that are fed these leaves will survive less well, and attain a smaller adult size,
than insects fed leaves from previously undamaged control trees.

TEST
Seventeen experimental birch trees were selected. Ten of these trees (chosen at random) were artificially defoliated
by scientists simulating an insect attack with scissors. The other seven trees were left as controls. In the following
year, twenty young insects were enclosed in a mesh bag on each tree, where they were allowed to feed and grow for
the rest of the season (about 5 weeks). Then the surviving insects (now pupae) were collected, counted, and weighed.
Average mass and survival was calculated for each tree, and then for each treatment (control trees and defoliated
trees; Table 1).

Table 1: Survival and pupal mass of caterpillars that were grown on control
trees and trees that were defoliated the previous season (mean ±standard error).
444444444444444444444444444444444444444444444444444444
Survival (%) Pupal mass (mg)
))))))))))))) )))))))))))))

Control trees (n = 7) 30 ± 3 58 ± 2

Defoliated trees (n = 10) 20 ± 4 48 ± 2

))))))))))))))))))))))))))))))))))))))))))))))))))))))

The probability of obtaining these results by chance alone (if we assume that there was no real difference between
control and defoliated trees) was less than one in a hundred for survival, and less than one in a thousand for pupal
weight. Therefore, we conclude that defoliated trees in general are of lower food quality for the caterpillars than
control trees.

FATE OF THE HYPOTHESIS

The prediction of the hypothesis was satisfied, therefore the hypothesis was supported.

THE ISSUE OF SUPPORT VS PROOF

Note that the hypothesis was NOT proven - it was supported. This is an important distinction. Other hypotheses
could also be consistent with these results. In fact, an appealing alternative hypothesis has been advanced by J.P.
Bryant, F.S. Chapin, and D.R. Klein, who argued that defoliated trees are nutrient stressed due to the loss of nitrogen
and phosphorus when leaves are consumed. As a result, the trees have a surplus of carbohydrates relative to that
which can be invested in growth, and as result, produce higher concentrations of carbon based secondary
metabolites, such as phenolics and tannins. Also, the leaves have lower nitrogen content. This would be similar to
what happens when you do not fertilize your lettuce patch and end up with a lousy salad.
Both Haukioja's defensive hypothesis and Bryant's nutrient stress hypothesis neatly explain the existing data. Either
one could be correct given the information we have. Yet the two hypotheses are very different: one describes a
surprisingly clever tree that is actively responding to the presence of insects by defending itself, and therefore
reducing the damage that it sustains; the other hypothesis describes a tree whose physiology and metabolism have
been disrupted by insect damage, with the accidental result that foliage quality for herbivores is reduced the next
year.
To discriminate between these competing hypotheses requires new predictions and new tests. Researchers were
challenged to devise experiments where the expected results are different depending on which hypothesis you
believe. For example, if the trees are actively defending themselves against herbivores, we might expect that they
respond more to real insect damage than they do to scientists simulating herbivory with scissors. There is no reason
to expect any such difference based on the nutrient stress hypothesis. In fact, Haukioja conducted this experiment
and found that real insect damage induced a stronger response. This result strengthened the defense hypothesis,
while simultaneously falsifying the nutrient stress hypothesis. But this was far from the last word.
Bryant, working on several species of trees in Alaska and Africa, obtained experimental results that conform much
better to the nutrient stress hypothesis than they do to the defense hypothesis. It now appears that there is some truth
in both explanations. As is common, the interesting question has become one of relative frequency. How many tree
species or tree populations behave as predicted by the defense hypothesis? How many behave as predicted by the
nutrient stress hypothesis? How many behave in a way predicted by neither? These questions cannot be reliably
answered at this time, but our understanding of tree "behavior" is much greater than it was a few years ago when
Haukioja first proposed his then radical defense hypothesis. Researchers in labs across the world are presently
developing imaginative new hypotheses, and devising and testing new predictions of existing hypotheses. It is in this
way that scientists continue to accumulate understanding about the nature of plant-insect interactions.

THE VALUE OF FALSIFYING A HYPOTHESIS

Consider the other possible result of Haukioja's defoliation experiment. Suppose that he had found no difference
between the control and defoliated trees. W ould this have been a useless result? W ould the experiment have been a
waste of time? No. In fact, if the hypothesis is interesting and realistic, if the prediction is carefully considered, and
the test is carefully conducted, then falsifying the hypothesis may be the most valuable result of all.
Note in Fig. 2 that experimental results supporting the hypothesis enters us into a potentially endless loop. Because
we cannot prove the hypothesis we must go back and test it again and again. For each test that supports the
hypothesis we become a bit more confident in it, but we will never obtain the single result that conclusively proves
it. Results that support a hypothesis are always somewhat open-ended. Contrast this with a result that does not
support the prediction. In this case we have a much more definite answer. The hypothesis is not correct. It has been
falsified, and we can proceed to formulate and test other hypotheses. W e have gained understanding by eliminating a
possible explanation. For example, if Haukioja's experiment had falsified the defense hypothesis, we would be quite
certain that inducible defense was not the explanation for how birch trees survive insect outbreaks, or for why insect
outbreaks are invariably followed by a crash. W e would now be more impressed with the importance of predators
and diseases in controlling insect populations, and we would be able to spend more time refining our understanding
of these remaining factors.
W hile a well conducted experiment adds to our understanding no matter what the result, it is easy to design
experiments that will only be useful with one of several possible results. This can be successful if you are lucky, but
all too often it will be a waste of time. Good scientists do not rely on being lucky. I briefly mention some of the
common pitfalls of ecological research and summarize the characteristics of a good experiment.

G OOD AND BAD SCIENTIFIC METHOD

THREE COMMON PITFALLS

! The experiment attempts to consider too many variables at once . Science proceeds in steps. It is always
tempting to answer everything at once, but confusion is the most likely result. In Haukioja's experiment, the
ONLY factor that differed between the two sets of trees was simulated insect damage. If, for example, the
defoliated trees had also been fertilized, then we would not know whether the difference in caterpillar
growth was due to defoliation or fertilization. W e would have learned less by trying to learn more.
! The hypothesis seems very improbable to everyone but you . This is a two-edged sword. If your hypothesis
turns out to be supported you may revolutionize thinking on the issue, but more likely the hypothesis will
be falsified, and no one will be the least bit surprised or benefited except you. It is not a good tactic to
depend upon obtaining any one particular result for your study to be interesting.
! The prediction does not necessarily follow from the hypothesis. Haukioja's experiment comparing real
insect damage with simulated insect damage falls into this category. Of course it turned out that Haukioja
was lucky, otherwise I would probably never have heard about the experiment. But suppose that he had
NOT found a difference between real and simulated damage. Birch trees could very well have been clever
enough to have an inducible defense system, yet not clever enough to tell the difference between insect
jaws and metal scissors. (They are, after all, just plants.) I doubt that Haukioja or anyone else would have
discarded the defense hypothesis based on a failure of that particular prediction.
CHARACTERISTICS OF A GOOD HYPOTHESIS
! Provides a reasonable explanation for the existing data
! Sound inductive reasoning
! Imaginative, unique
! Has interesting and important implications
! Is testable
! Not necessarily correct

Hypothesis formulation is a very subjective aspect of the scientific method. Besides being testable and logically
sound, good hypotheses are imaginative and unique. They provide a previously unexplored explanation. Haukioja
was willing to imagine a clever, active, birch tree, which seemed preposterous at the time. However, so much
support has since accumulated for highly organized, inducible defenses in plants, that it is now a standard
expectation for ecologists, farmers, plant-breeders, foresters, etc.

CHARACTERISTICS OF A GOOD PREDICTION

! Sound deductive reasoning
! Closely follows the hypothesis
! Not easily arrived at by other competing hypotheses

CHARACTERISTICS OF A GOOD TEST

! Repeatable
! Unambiguous
! Suitably powerful
The test itself may be the only component of the scientific method that is entirely objective. The outcome of a test
should not be influenced by the hopes of the researcher. A good test should be repeatable at other times and by other
scientists. The results should be clear in that the predictions are unambiguously supported or not. A good test should
contain a large enough sample size (10 trees and 7 trees in Haukioja's example) to reliably detect differences that are
of biological interest. If Haukioja had used only 2 trees in each treatment, but still obtained the same average values
for survival and pupal weight, he would have been unable to convince us that defoliated trees in general differ from
control trees. This is an issue that can be treated formally using statistics, but even without statistics training, you
should have a sense for the value of replication in experiments. Large sample sizes can however become too much of
a good thing. If Haukioja had used 200 trees instead of 17 then it would have taken him dozens of man-years to
actually conduct the experiment (defoliating trees, weighing pupae, etc.). This enormous sample size would have
allowed him to demonstrate that even tiny differences were statistically significant. But statistically significant
differences are not necessarily biologically meaningful. W ith a sample of 200 trees, 20% survival could differ
significantly from 21% survial, but it would still be unlikely to materially affect the population size of the insects.

REFERENCES
Bryant, J.P., F.S. Chapin, and D.R. Klein. 1983. Carbon/nutrient balance of boreal plants in relation to vertebrate
herbivory. Oikos 40: 357-368.
Haukioja, E., and S. Neuvonen. 1985. Induced long-term resistance of birch foliage against defoliators: tests of
defensive and non-defensive hypotheses. Ecology 66:1303-1308
Haukoja, E., J. Suomela, and S. Neuvonen. 1985. Long-term inducible resistance in birch foliage: triggering cues
and efficacy on a defoliator. Oecologia 65: 363-369.
Bloom AJ, Chapin FS, III, Mooney HA. 1985. Resource limitation in plants-an economic analogy. Annual Review
of Ecology and Systematics 16:363-392.