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UNIT PROJECT WORKSHEET

Case Study: Malformed Frogs
Follow a real-life mystery in this frog case study. Work with a partner or a small group to read and
discuss the facts of this unsolved case. At the end of the case study, you will conduct your own
research based on what you have learned. Use your Evidence Notebook to record your ideas as the
case develops. Maybe you can find the clue that solves this case!

LESSON 1: DNA STRUCTURE AND REPLICATION

It all started with a class field trip. In August 1995, students from the Minnesota New Country School
in Le Sueur went on a field trip to study the wetlands. On a farm in Henderson near the Minnesota
River, the students saw frogs in Ney Pond. But something was wrong.

“The students from the Minnesota New Country School looked at 22 frogs,” said teacher Cindy
Reinitz, “half of which were deformed.” The northern leopard frogs in Ney Pond had malformed
limbs. Some frogs had contorted limbs. Some frogs had shortened limbs. Some frogs had extra legs
growing on their legs while others had missing limbs. This was not a stage in the life cycle of a frog
that the students expected to see.

Judy Helgen, a biologist at the Minnesota Pollution Control Agency (MPCA), was called to the
scene. “It was terrible. The kids and the parents, everybody looked really worried,” said Helgen.
“The kids had already collected some frogs, and then we went around and did some surveying,
and it was horrible.”

Address the following in your Evidence Notebook:

1. Predict Use Internet resources to find images of the mutated frogs the students found at
Ney Pond in Henderson, Minnesota. After viewing the images, answer the following
questions: How do you think DNA structure and function might be related to the development
of malformed limbs in these frogs? Do you think the frogs inherited these mutations from their
parents, or are they the result of other factors?

2. Describe factors you would want to investigate if you were a scientist in Minnesota in August
1995. Write a hypothesis predicting how one of these factors might affect frog malformations.
Then, write a list of steps you could take to test your hypothesis.

3. Research the life cycle of frogs. Draw and label a diagram illustrating the life cycle of a frog.
Then, answer this question: At which stages of a frog’s development do you think it is most
vulnerable to the factors that might have caused these malformations? Explain your answer.

LESSON 2: PROTEIN SYNTHESIS

What was causing these frog malformations? The only way to find out was to investigate. The
students from Minnesota New Country School who made the alarming discovery continued to collect
frogs for the researchers to study. “Within a month they had collected roughly 400 deformed frogs
from the Ney Pond,” said teacher Cindy Reinitz.

News of the students’ deformed frogs discovery was reported in the media around the world. The
Minnesota state legislature set up a citizens frogwatch patrol and hotline. “We had a call from this
teacher in Litchfield,” said biologist Judy Helgen. “She had a kid bring in a bucket of deformed frogs
to school.” But the calls didn't stop there. “It became sort of chilling,” Helgen said, “because we
started getting phone calls, and not just from Minnesota, but from people in other states.”

Unit Project Worksheet 1 The Structure and Function of DNA

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In 1996, the MPCA collected data from across the state. Frogs with missing or extra legs were
found on farms and in neighborhoods at almost 150 ponds and lakes. “Worldwide, nobody has ever
reported deformities on this level. Things have just gone crazy. They’ve just exploded, and it’s the
suddenness of it,” said Ralph Pribble, the Public Information Offier at the Minnesota Pollution
Control Agency.

The MPCA asked a professor at the University of Minnesota for help. Geneticist and biology teacher
Bob McKinnell had studied nothern leopard frogs and cancer for many years. “I had collected
extensively in South Dakota, North Dakota, Minnesota, and in years previous to that in Vermont,”
said McKinnell. “And I simply had never, never, never seen the abundance of abnormalities such as
Judy and those students were seeing in Henderson and subsequently throughout Minnesota and
even throughout the world.”

The MPCA was the first government agency on the case, but they were’t the only ones investigating
possible causes for the frog malformations. As more malformed frogs were found, more scientists
began to study the problem and write about their findings. At one point, scientists from a number of
universities and federal agencies (National Institute of Environmental Health Sciences, U.S. Fish and
Wildlife Service, U.S. Geological Survey, and U.S. Department of Agriculture) were also working on
the problem.

The scientists’ research was summarized in a U.S. Geological Survey field guide in 2000. The
students in Minnesota (and the public) wanted to know what went wrong. How could this happen?
What caused malformations? In the 2000 Field Guide to Malformations of Frogs and Toads With
Radiographic Interpretations, Carol U. Meteyer explained, “Malformations represent primary errors in
development, errors in chemical communication or translation of genetic information.”

Address the following in your Evidence Notebook:

1. Use Internet resources to locate the Minnesota Pollution Control Agency Malformed Frogs
Database, hosted by the U.S. Geological Survey. Print out or take screenshots of maps and
data tables that you find, and use them to answer these questions: What patterns do you
notice in the data? What do these trends tell you about the factors that might have caused
the malformations in these frogs?

2. Use Internet resources to locate the Field Guide to Malformations of Frogs and Toads with
Radiographic Interpretations. After viewing the images there, answer the following questions:
How do you think protein synthesis might be related to the malformations in the frogs? Draw
or describe a model to explain how protein synthesis may contribute to the malformations in
the frogs’ limbs.

3. Predict How could you study whether the model you described in Question 2 is correct?
How do you think scientists study protein synthesis and other processes that occur at the
molecular level in an organism?

LESSON 3: GENE EXPRESSION AND REGULATION

“Frogs are very sensitive to small amounts of outside agents in their environment, so they are good
monitors of that environment,” said University of Minnesota geneticist Bob McKinnell. “Those
abnormalities are a red flag, but until we study it more, no one can say it’s a warning sign for
anything except for the frogs themselves.”

Solving a biological mystery is complicated. The malformed frogs that the students and the
researchers found were the survivors. No one knew how many frogs had already died. To find the
source of the problem, scientists needed to conduct experiments at the earliest stages of frog
development. They had to begin with frog embryos and tadpoles.

Unit Project Worksheet 2 The Structure and Function of DNA

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Deciding what to test was the next obstacle. “Minnesota Pollution Control Agency staff and other
researchers are testing the mud, water, and frog tissues for abnormal levels of metals such as
arsenic, mercury, selenium, and cadmium,” said Judy Helgen, a biologist at the MPCA. “They’re also
looking for chemicals like PCBs, herbicides, and pesticides, and also at parasites as potential
causes.” No one knew what was causing the malformations, so it was much too early to rule
anything out, even heredity. “Researchers speculate the deformities may be the result of genetic
mutations passed on from the parent frogs to their offspring,” said Helgen.

A CRACK IN THE CASE

In the spring of 1999, there was a crack in the case. The American Association for the Advancement
of Science sent out a press release stating, “[The] two studies in the 30 April issue of Science
provide the strongest evidence yet that at least some of the frogs’ bizarre deformities are the direct
result not of pesticides or ozone loss, but of a parasite that naturally infects the animals.”

One study, which Stanley Sessions began in 1987, showed that “small parasitic flatworms called
Riberoria trematodes” were the culprit. When trematodes burrowed into a tadpole to use it as a host,
the flatworms rearranged the tadpole’s hind legs. The frog host grew multiple legs. “It’s about as
close to using an egg beater on the limb bud cells as you can get,” said Sessions. He built his
1987–1999 study on an earlier study of vitamin A derivatives, called retinoids. In the earlier study,
scientists disovered that retinoids caused multiple limbs to develop. A result from one study led to
the idea for another.

The second study reported in the April 30, 1999, issue of Science confirmed Sessions’s trematode
studies. Pieter Johnson began studying tadpoles and trematodes for his honors graduate thesis at
Stanford University. Johnson and his collegues tested tadpoles in water with varying amounts of the
parasitic trematodes. “We found a high frequency of deformities even [under conditions with] low
parasite density,” said Johnson.

ONE PIECE OF THE PUZZLE

Was the case solved? Not quite. “We’ve chipped off the most mysterious-looking piece of the
deformed limb puzzle,” said Sessions. These two studies showed that trematode infections were the
reason that frogs grew two or three properly formed legs from one limb bid. But that was just one
piece of the puzzle. What was responsible for the other malformations? Why were some frogs
growing extra legs that weren't properly formed? What caused some frogs to develop without their
limbs or their eyes? No one knows, yet . . .

Address the following in your Evidence Notebook:

1. The scientists in this case study mentioned several external factors that can influence gene
expression in frogs, such as metals, pesticides, herbicides, and parasites. Choose one of
these factors to research further, and summarize your findings. What can you conclude about
how this factor affects gene expression in frogs?

2. Predict Make a model that illustrates how the factor you chose may affect gene expression
in frogs and how these changes might lead to limb malformations.

CONCLUSION
Apply what to have learned in the case study and conduct your own investigation using these steps.

1. Explain your prediction, and write a hypothesis.

2. Describe the procedure you followed as you conducted your research. Name each source
you researched, and explain what the source revealed. Include any answers the source
provided and any new questions that arose after consulting that source.

3. After you collect data from three to five sources, analyze the data and draw a conclusion. Did
your findings support your hypothesis?

Unit Project Worksheet 3 The Structure and Function of DNA

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EXTEND
Gather Evidence Investigate another animal affected by environmental toxins. Select an animal
that lives in your area, or study the effect of DDT on birds, such as the brown pelican and the bald
eagle. For a case study approach, begin with the book that began the environmental movement in
1962, Silent Spring by Rachel Carson. Select one of the animals chronicled in the book, and
research what has happened between then and now.

PRESENT
Make a poster or a computer presentation that answers these questions:

1. Why are proteins so important in the functioning of a body?

2. How are proteins related to DNA?

3. What can cause damage or a change in DNA, and how does that change the structure and
function of proteins?

Use evidence gathered over the course of your research and discussion to support your claims.
Photos and drawings may be useful evidence to include on your poster. Your poster or presentation
should also include your data tables and graphs.

Unit Project Worksheet 4 The Structure and Function of DNA

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