Brain-Eating Ameba
Developed by
Jewel B. Moses, PhD Arcot M. Saibaba, PhD
Southeast Middle School Newton High School
Hopkins, South Carolina Covington, Georgia
This lesson plan was developed by teachers attending the Science
Ambassador Workshop. The Science Ambassador Workshop is a career
workforce training for math and science teachers. The workshop is a Career
Paths to Public Health activity in the Division of
Scientific Education and
Professional Development, Center for Surveillance, Epidemiology,
and Laboratory Services, Office of Public Health Scientific Services,
Centers for Disease Control and Prevention.
CDC Science Ambassador Workshop
2013 Lesson Plan
Acknowledgements
This lesson plan was developed in consultation with subject matter experts from the Division of
Foodborne, Waterborne, and Environmental Diseases, National Center for Emerging and Zoonotic
Infectious Diseases, Centers for Disease Control and Prevention.
Jennifer Cope, MD, MPH
Medical Epidemiologist
Jonathan Yoder, MSW, MPH
Epidemiologist
Scientific and editorial review was provided by Ralph Cordell, PhD and Kelly Cordeira, MPH from
Career Paths to Public Health, Division of Scientific Education and Professional Development, Center
for Surveillance, Epidemiology, and Laboratory Services, Office of Public Health Scientific Services,
Centers for Disease Control and Prevention.
Suggested citation
Centers for Disease Control and Prevention (CDC). Science Ambassador Workshop—Brain-eating
Ameba. Atlanta, GA: US Department of Health and Human Services, CDC; 2013. Available at
http://www.cdc.gov/scienceambassador/lesson-plans/index.html.
Contact Information
Please send questions and comments to scienceambassador@cdc.gov.
Disclaimers
This lesson plan is in the public domain and may be used without restriction.
Citation as to source, however, is appreciated.
Links to nonfederal organizations are provided solely as a service to our users. These links do
not constitute an endorsement of these organizations nor their programs by the Centers for
Disease Control and Prevention (CDC) or the federal government, and none should be
inferred. CDC is not responsible for the content contained at these sites. URL addresses listed
were current as of the date of publication.
Use of trade names and commercial sources is for identification only and does not imply
endorsement by the Division of Scientific Education and Professional Development, Center
for Surveillance, Epidemiology, and Laboratory Services, CDC, the Public Health Service, or
the U.S. Department of Health and Human Services.
The findings and conclusions in this Science Ambassador Workshop lesson plan are those of
the authors and do not necessarily represent the official position of the Centers for Disease
Control and Prevention (CDC).
Contents
Summary ................................................................................................................................................. 1
Learning Outcomes ................................................................................................................................. 2
Duration .................................................................................................................................................. 2
Procedures ............................................................................................................................................... 3
Day 1: Introduction to Parasites, 45 minutes ........................................................................................ 3
Preparation ......................................................................................................................................... 3
Materials ............................................................................................................................................. 3
Online Resources ............................................................................................................................... 3
Activity ............................................................................................................................................... 4
Day 2: Parasitic Environments, 45 minutes ......................................................................................... 5
Preparation ......................................................................................................................................... 5
Materials ............................................................................................................................................. 5
Online Resources ............................................................................................................................... 5
Activity ............................................................................................................................................... 6
Conclusion .............................................................................................................................................. 7
Assessments ............................................................................................................................................ 7
Educational Standards ............................................................................................................................. 8
List of Appendices ................................................................................................................................. 10
Appendix 1A: Waterborne Disease Case Study, Scenario Worksheet ................................................ 11
Appendix 1B: Waterborne Disease Case Study, Scenario Worksheet Answer Key .......................... 12
Appendix 2A: Waterborne Disease Case Study, Graphic Organizer .................................................. 13
Appendix 2B: Waterborne Disease Case Study, Graphic Organizer Answer Key .............................16
Appendix 3A: Waterborne Disease Case Study, Laboratory Exploration ......................................... 19
Appendix 3B: Waterborne Disease Case Study, Laboratory Exploration Answer Key .....................22
Appendix 4: Waterborne Disease Case Study, Public Awareness Campaign ..................................... 26
Appendix 5A: Brain-Eating Ameba Quiz .......................................................................................... 27
Appendix 5A: Brain-Eating Ameba Quiz, Answer Key .................................................................... 28
1
Brain-Eating Ameba
Summary
Swimming is a fun summer activity, but it can
increase your risk for contracting certain waterborne
diseases. Naegleria fowleri (N. fowleri) is among
many organisms that live in recreational waters, such
as ponds and lakes, and is commonly referred to as
the brain-eating ameba. It is a single-celled,
bacteria-eating organism that can be found in warm
fresh water around the world. It is a free-living
ameba that lives in the environment and does not
need a human host to complete its life cycle.
Fortunately, infections caused by N. fowleri are rare
and the risk for diseases is very low. During the
summer of 2013, only four cases were reported in
the United States and during the 10 years from 2005
to 2014, a total of 35 cases were reported, despite
millions of recreational water exposures each year.
By comparison, during the 10 years from 2001 to
2010, >34,000 drowning deaths occurred in the
United States. N. fowleri usually infects people when
contaminated water enters the body through the
nose. You cannot become infected from swallowing
water contaminated with N. fowleri.
This lesson plan demonstrates how microorganisms
normally found in environments, such as the bottom
of warm freshwater ponds and lakes can cause illness
when they enter the human body. Students engaged in
this lesson plan will learn about N. fowleri (the
scientific name of the brain-eating ameba), where it
lives, how it can cause infection, and how persons can
protect themselves from this infection. Students will
also have the opportunity to identify other organisms
living in local freshwater reservoirs, such as ponds
and lakes. At the end of the lesson, students should
have an enhanced understanding of the environment’s
role in disease transmission and ways to reduce the risk for contracting waterborne infections.
This material is suitable for use in high school biology or environmental science classes and can be
included as part of lessons on aquatic ecosystems. Before studying this lesson, students should have a
basic understanding of the following six kingdom classifications: Archaebacteria, Eubacteria, Protista,
Fungi, Plantae, and Animalia. Student prior knowledge of anatomy of the human nose and brain would
also be helpful.
Figure 1. Naegleria fowleri has 3 stages in its life cycle:
cysts, ameboid trophozoites, and flagellates. The
only infective stage of the ameba is the ameboid trophozoite.
Trophozoites are 1035 µm long with a granular appearance
and a single nucleus. The trophozoites replicate by binary
division during which the nuclear membrane remains intact (a
process called promitosis) Trophozoites infect humans or
animals by penetrating the nasal tissue and migrating to
the brain via the olfactory nerves causing primary amebic
meningoencephalitis (PAM).
Source:
http://www.cdc.gov/parasites/naegleria/pathogen.html
2
Learning Outcomes
After completing this lesson, students should be able to
explain how freshwater organisms, such as the brain-eating ameba (N. fowleri), are affected by
environmental variations (e.g., temperature, nutrient availability, geography, and life-cycle stages);
describe limitations of epidemiologic and laboratory evidence; and
use critical thinking to construct an evidence-based explanation as to why an N. fowleri infection
might occur.
Duration
This lesson can be conducted as one 90-minute lesson or divided into two 45-minute lessons.
3
Procedures
Day 1: Introduction to Parasites, 45 minutes
Preparation
Before Day 1,
Review the Centers for Disease Control and Prevention (CDC) Internet site regarding N. fowleri. See
the Online Resources section that follows.
Schedule computer laboratory or library time or reserve student laptop cart.
For the Waterborne Disease Case Study, make one copy per student of Appendix 1A, Scenario
Worksheet; and Appendix 2A, Graphic Organizer.
Collect water samples from a local untreated water source (preferably a lake or a pond) or prepared
slides of organisms that would be in lake or pond water, including protista (flagellates, amebae
heliozoans, and ciliates), bacteria, algae, rotifers, hydra, worms, and arthropods. Students will use
these water samples on Day 2 to prepare wet mounts or use prepared slides for microscopic analysis.
Note: Surface water often has a relatively low concentration of organisms; because you likely will
require a higher concentration to be able to see much with a light microscope, try to collect the
sample from a greater depth, preferably near the bottom. Alternatively, collect prepared slides of
organisms that would be in lake or pond water, including protista (flagellates, amebae, heliozoans,
and ciliates), bacteria, algae, rotifers, hydra, worms, and arthropods.
Materials
Appendix 1A, Scenario Worksheet
Description: This case study engages student curiosity as they investigate the cause of infection in a
male aged 14 years.
Appendix 2A, Graphic Organizer
Description: Students use this worksheet to organize their notes.
Computers with Internet access.
Online Resources
CDC Internet site http://www.cdc.gov
Description: To determine the correct pathogen responsible for the victim’s death in the case study,
students research the three ameba options (Dientamoeba fragilis, Acanthamoeba, and N. fowleri) by
entering the terms into CDC’s search function.
CDC Internet site on N. fowleri: http://www.cdc.gov/parasites/naegleria/index.html
Description: Students will use this website to complete their graphic organizer.
Matthews S, Ginzl D, Walsh D, et al. Primary amebic meningoencephalitis—Arizona, Florida, and
Texas, 2007. MMWR Morbidity and Mortality Weekly Report 2008;57:573–7.
URL: http://www.cdc.gov/mmwr/preview/mmwrhtml/mm5721a1.htm?s_cid=mm5721a1_eURL.
Description: The original case study of an Arizona male aged 14 years infected with N. fowleri.
4
Activity
1. Read the Appendix 1A scenario aloud. Challenge students to determine the ameba (Dientamoeba
fragilis, Acanthamoeba, or N. fowleri) responsible for the infection described in the case study by
using http://www.cdc.gov and other search functions. After students discover that N. fowleri is
responsible for the infection, pose the question, “What are some dangers of swimming in untreated
water? Talking points for this discussion can include
a. N. fowleri infections can be serious, but they are rare; although there is no need to be scared
of swimming in local lakes and ponds to avoid N. fowleri, it is best to practice simple
prevention measures, such as keeping head above water or using nose clips.
b. More common dangers related to swimming in fresh water include
infections, such as leptospirosis, swimmer’s itch (cercarial dermatitis), giardiasis,
cryptosporidiosis, cholera, shigellosis, and norovirus and infections of Escherichia
coli, including E. coli O157:H7;
wound infections;
diving hazards and other water-related injuries; and
drowning.
c. The majority of infections and dangers related to swimming in fresh water can be prevented.
2. Assign Appendix 2A, Graphic Organizer. Direct students to research N. fowleri environment,
nutrients, portal of entry into the human host, and patient symptoms using the CDC Internet site on
N. fowleri at http://www.cdc.gov/parasites/naegleria/index.html.
5
Day 2: Parasitic Environments, 45 minutes
Preparation
Before Day 2,
Review Internet resources and microscope procedures and safety.
Make copies of Appendix 3A, Laboratory Exploration; Appendix 4, Public Awareness Campaign;
and Appendix 5A, Brain-Eating Ameba Quiz.
Schedule computer laboratory or library time or reserve student laptop cart.
Retrieve water samples collected on Day 1 and prepare microscopes for the laboratory.
Materials
Appendix 5A, Brain-Eating Ameba Quiz
Description: This test gauges student knowledge of the brain-eating ameba.
Water samples from a local untreated water source (preferably a lake or a pond) or prepared slides of
organisms that would be in lake or pond water, including protista (flagellates, amebae, heliozoans,
and ciliates), bacteria, algae, rotifers, hydra, worms, and arthropods
Description: Students use these water samples to prepare wet mounts or will use prepared slides for
microscopic analysis.
Compound microscopes
Laboratory gloves
Computers with Internet access
Appendix 3A, Laboratory Exploration
Description: This worksheet is used for the laboratory activity.
Appendix 4, Public Awareness Campaign
Description: This worksheet is used for the laboratory activity.
Online Resources
Pond Life Identification Kit
URLs:
http://www.microscopy-uk.org.uk/index.html?http://www.microscopy-uk.org.uk/pond/index.html
and
http://www.microscopy-uk.org.uk/index.html?http://www.microscopy-
uk.org.uk/pond/protozoa.html.
Description: These Internet sites provide images and descriptions of key features of certain common
freshwater organisms, including protista.
Recreational Water Illnesses and Prevention Tips
URL: http://www.cdc.gov/healthywater/swimming/rwi/illnesses/index.html.
Description: This Internet site contains information regarding recreational water illnesses and how to
avoid them.
6
Activity
1. Assign students Appendix 5A, Brain-Eating Ameba Quiz. Students should complete the short quiz
on their own and then review the answers as a class.
2. Review laboratory and microscope procedures, including microscope parts, magnification, and
focusing techniques; safe handling of the water samples; and wet-mount preparation.
3. Divide the class into pairs or groups of 3 for the microscope activity. Have each group analyze the
water sample assigned to them and identify organisms living in the sample. Have students draw each
organism on Appendix 3C, Laboratory Exploration. Using the Pond Life Identification Kit, have
students determine the scientific name of each organism.
4. Ask students to state the most common organism group found in their sample and to estimate the
number of amebae found in their sample. As a class, discuss potential reasons why the number of
amebae differs from group to group. Use this example to discuss why no method exists that
accurately and reproducibly measures the number of amebae in the water.
5. Prompt students to answer questions 35 in their group. Discuss the answers.
6. For homework, instruct students to design a public health awareness campaign using a most-wanted-
type poster or a brochure that illustrates how to reduce the risk for acquiring their assigned
waterborne disease. See Appendix 4.
7
Conclusion
Students use a public health scenario to learn about limnology, microbiology, and public health. By
examining the life cycle of N. fowleri, students examine how environmental variation in the lake, such
as temperature, nutrient availability, and geography, can affect the life cycle of an organism. Then,
students learn how life-cycle changes can increase the risk for human infection leading to the disease
called primary amebic meningoencephalitis. However, since no data exist to accurately estimate the true
risk, students use critical thinking to formulate hypotheses that account for limitations in epidemiology
and the laboratory.
Assessments
Appendix 1A, Scenario Worksheet
Learning Outcomes Assessed:
- Explain how freshwater organisms, such as the brain-eating ameba (N. fowleri), are affected by
environmental variations (e.g., temperature, nutrient availability, geography, life-cycle stages).
- Describe limitations of epidemiological and laboratory evidence.
- Use critical thinking to construct an evidence-based explanation as to why a case of brain-eating
infection may occur.
Description: This case study engages student curiosity as they investigate the cause of infection in a
male aged 14 years.
Appendix 5, Brain-Eating Ameba Quiz
Learning Outcome Assessed:
- Explain how freshwater organisms, such as the brain-eating ameba (N. fowleri), are affected by
environmental variations (e.g., temperature, nutrient availability, geography, and life-cycle
stages).
Description: This quiz of five multiple-choice questions gauges student knowledge of the brain-
eating ameba and should take approximately 10 minutes to complete.
8
Educational Standards
In this lesson, the following CDC Epidemiology and Public Health Science (EPHS) Core Competencies
for High School Students
1
, Next Generation Science Standards
*
(NGSS) Science & Engineering
Practices
2
, and NGSS Cross-cutting Concepts
3
are addressed:
HS-EPHS1-2. Discuss how epidemiologic thinking and a public health approach is used to transform a
narrative into an evidence-based explanation.
NGSS Key Science & Engineering Practice
2
Obtaining, Evaluating and Communicating Information
Critically read scientific literature adapted for classroom use to determine the central ideas or
conclusions and/or to obtain scientific and/or technical information to summarize complex
evidence, concepts, processes, or information presented in a text by paraphrasing them in
simpler but still accurate terms.
NGSS Key Crosscutting Concept
3
Cause and Effect
Empirical evidence is required to differentiate between cause and correlation and make
claims about specific causes and effects.
HS-EPHS2-4. Use patterns in empirical evidence to formulate hypotheses.
NGSS Key Science & Engineering Practice
2
Asking Questions & Defining Problems
Ask questions that arise from careful observation of phenomena, or unexpected results, to
clarify and/or seek additional information, that arise from examining models or a theory, to
clarify and/or seek additional information and relationships, to determine relationships,
including quantitative relationships, between independent and dependent variables, and to
clarify and refine a model, an explanation, or an engineering problem.
NGSS Key Crosscutting Concept
3
Patterns
Empirical evidence is needed to identify patterns.
HS-EPHS3-5. Make a claim about causality with consideration of a mathematical analysis of empirical
data and Bradford Hill’s Criteria for Causality.
NGSS Key Science & Engineering Practice
2
Engaging in Argument from Evidence
Make and defend a claim based on evidence about the natural world or the effectiveness of a
design solution that reflects scientific knowledge and student-generated evidence.
NGSS Key Crosscutting Concept
3
Systems and System Models
Models can be used to predict the behavior of a system, but these predictions have limited
precision and reliability due to the assumptions and approximations inherent in models.
*
Next Generation Science Standards is a registered trademark of Achieve. Neither Achieve nor the lead states and partners
that developed the Next Generation Science Standards was involved in the production of, and does not endorse, this product.
9
1
Centers for Disease Control and Prevention (CDC). Science Ambassador WorkshopEpidemiology and Public Health
Science: Core Competencies for high school students. Atlanta, GA: US Department of Health and Human Services, CDC;
2015. Not currently available for public use.
2
NGSS Lead States. Next Generation Science Standards: For States, By States (Appendix FScience and Engineering
Practices). Achieve, Inc. on behalf of the twenty-six states and partners that collaborated on the NGSS. 2013. Available at:
http://www.nextgenscience.org/sites/ngss/files/Appendix%20F%20%20Science
%20and%20Engineering%20Practices%20in%20the%20NGSS%20-%20FINAL%20060513.pdf
3
NGSS Lead States. Next Generation Science Standards: For States, By States (Appendix GCrosscutting Concepts).
Achieve, Inc. on behalf of the twenty-six states and partners that collaborated on the NGSS. 2013. Available at:
http://www.nextgenscience.org/sites/ngss/files/Appendix%20G%20-
%20Crosscutting%20Concepts%20FINAL%20edited%204.10.13.pdf.
.
10
Appendices: Supplementary Documents
11
Appendix 1A
Waterborne Disease Case Study
Scenario Worksheet
Name: __________________________________ Date: ________________
Directions: Read the following case study on a waterborne disease. Next, use the CDC Internet site
(http://www.cdc.gov) to determine if the ameba responsible for the victim’s death might have been
Dientamoeba fragilis, Acanthamoeba, or Naegleria fowleri.
In Arizona, an adolescent male aged 14 years was enjoying the last of his summer days swimming in a
northeastern Arizona lake. He was observed diving and splashing in shallow water. The water
temperature on that day, September 8, was 86.3°F (30.2°C), and the air temperature was 108.0°F
(42.2°C). Approximately 1 week later, on September 14, he experienced a severe headache, stiff neck,
and fever. On September 16, he was hospitalized with possible meningitis (inflammation of the
meninges of the brain). From a spinal tap, doctors observed an ameba in his cerebrospinal fluid by using
a microscope. Note: The meninges are three layers of tissue that surround the brain and spinal cord to
protect the central nervous system.
1. What is the name of the ameba responsible for the victim’s death?
2. How might have infection occurred?
3. List at least three factors that led you to your answer in Question 1.
12
Appendix 1B
Waterborne Disease Case Study
Scenario Worksheet Answer Key
Name: ___________________________________ Date: ________________
Directions: Read the following case study on a waterborne disease. Next, use the CDC Internet site
(http://www.cdc.gov) to determine if the ameba responsible for the victim’s death might have been
Dientamoeba fragilis, Acanthamoeba, or Naegleria fowleri.
In Arizona, an adolescent male aged 14 years was enjoying the last of his summer days swimming in a
northeastern Arizona lake. He was observed diving and splashing in shallow water. The water
temperature on that day, September 8, was 86.3°F (30.2°C), and the air temperature was 108.0°F
(42.2°C). Approximately 1 week later, on September 14, he experienced a severe headache, stiff neck,
and fever. On September 16, he was hospitalized with possible meningitis (inflammation of the
meninges of the brain). From a spinal tap, doctors observed an ameba in his cerebrospinal fluid by using
a microscope. Note: The meninges are three layers of tissue that surround the brain and spinal cord to
protect the central nervous system.
1. What is the name of the ameba responsible for the victim’s death?
Answer: Naegleria fowleri. Note: Students should be able to deduce which ameba was responsible
by the characteristics given in the scenario and the information provided on the CDC Internet site:
http://www.cdc.gov/. For more details, see question 3.
2. How might have infection occurred?
Answer: Infection might have occurred as a result of swimming in the northeastern Arizona lake.
Students can also consider some of the other possible sources before coming to this conclusion,
including bodies of warm freshwater, such as lakes and rivers; geothermal (naturally hot) water, such
as hot springs; warm water discharge from industrial plants; geothermal (naturally hot) drinking
water sources; swimming pools that are poorly maintained, minimally chlorinated or unchlorinated;
water heaters; and soil.
3. List at least three factors that led you to your answer in Question 1.
Answer: Explanations might include the following: infection might have occurred as a result of
swimming in a lake in a U.S. state with a high number of case reports (5–9 cases); the optimal water
temperature range for the growth of the brain-eating ameba is 45°C–55ºC; average time of symptom
onset is typically 5 days (range, 1–7 days); average time from symptom onset to death is 5.3 days
(range, 1–12 days); early signs and symptoms include fever and headache; later signs and symptoms
include neck stiffness and seizures; amebae in the trophozoite morphology were observed in the
spinal fluid.
13
Appendix 2A
Waterborne Disease Case Study
Graphic Organizer
Name: __________________________________ Date: ________________
Directions: Complete the case study graphic organizer.
After Naegleria fowleri (N. fowleri) was found in trophozoite form in the spinal fluid, the boy was
diagnosed with primary amebic meningoencephalitis. To become more familiar with this waterborne
disease, use this worksheet to organize your notes into five sections: (1) scientific name and life-cycle
overview, (2) the cause-and-effect association between the environment and parasites, (3) diagnostics
and treatment strategies, (4) surveillance and epidemiology, and (5) risk communication challenges. Use
N. fowleri as an example in each section. Use the CDC Internet site on N. fowleri at
http://www.cdc.gov/parasites/naegleria/index.html to find information; take notes with a partner.
Scientific name and life-cycle overview
What is the common name for N. fowleri?
Describe or illustrate the life cycle of the brain-eating ameba.
14
Cause-and-effect association between the environment and parasites
How can the environment affect a parasite’s life cycle?
State examples of environmental factors that can affect a parasite’s life cycle:
What is the optimal temperature for growth of the brain-eating ameba?
How does the brain-eating ameba cause disease?
What is primary amebic meningoencephalitis (PAM)?
15
Diagnostics and treatment strategies
What are some diagnostic strategies for brain-eating amebic infection and PAM?
What are the treatment options for brain-eating ameba infection/PAM?
Surveillance and epidemiology
How many cases of PAM were reported during 1962–2014?
Who is most likely to be infected with this brain-eating ameba?
When is the most common time of the year to become infected with this brain-eating ameba?
Where is exposure to the brain-eating ameba most likely to occur?
Environmental prevention challenges
What are certain environmental concerns regarding N. fowleri infection?
16
Appendix 2B
Waterborne Disease Case Study
Graphic Organizer Answer Key
Name: __________________________________ Date: ________________
Directions: Complete the case study graphic organizer.
After Naegleria fowleri (N. fowleri) was found in trophozoite form in the spinal fluid, the boy was
diagnosed with primary amebic meningoencephalitis. To become more familiar with this waterborne
disease, use this worksheet to organize your notes into five sections: (1) scientific name and life-cycle
overview, (2) the cause-and-effect association between the environment and parasites, (3) diagnostics
and treatment strategies, (4) surveillance and epidemiology, and (5) risk communication challenges. Use
N. fowleri as an example in each section. Use the CDC Internet site on N. fowleri at
http://www.cdc.gov/parasites/naegleria/index.html to find information; take notes with a partner.
Scientific name and life-cycle overview
What is the common name for N. fowleri?
Answer: Brain-eating ameba
Describe or illustrate the life cycle of the brain-eating ameba.
Answer: N. fowleri has three stages in its life cycle: ameboid trophozoites, flagellates, and cysts. The
only infective stage of the ameba is the ameboid trophozoite. Trophozoites are 10–35 µm long, with a
granular appearance and a single nucleus. The trophozoites replicate by binary division during which
the nuclear membrane remains intact (a process called promitosis). Trophozoites infect humans or
animals by penetrating the nasal tissue and migrating to the brain through the olfactory nerves,
causing primary amebic meningoencephalitis (PAM).
Source: http://www.cdc.gov/parasites/naegleria/pathogen.html
17
Cause-and-effect association between the environment and parasites
How can the environment affect a parasite’s life cycle?
Answer: Trophozoites can turn into a temporary, nonfeeding, flagellated stage (10–16 µm long) when
stimulated by adverse environmental changes, such as a reduced food source. If the environment is
not conducive to continued feeding and growth (e.g., cold temperatures and scarce food), the ameba
or flagellate forms a cyst. The cyst form is spherical and 7–15 µm in diameter. It has a smooth, single-
layered wall with a single nucleus. Cysts remain resistant to environmental stresses to increase their
chance of survival until better conditions occur.
Source: http://www.cdc.gov/parasites/naegleria/pathogen.html
State examples of environmental factors that can affect a parasite’s life cycle:
Answer: Examples include cold temperatures or food scarcity.
What is the optimal temperature for growth of the brain-eating ameba?
Answer: N. fowleri grows best at higher temperatures up to 115°F (46°C). Although the ameba might
not be able to grow well, N. fowleri can still survive at even higher temperatures for limited periods.
The trophozoites and cysts can survive from minutes to hours at 122°F–149°F (50°C–65°C), with the
cysts being more resistant to these higher temperatures. Although trophozoites are killed rapidly by
refrigeration, cysts can survive for weeks to months at cold temperatures, but they appear to be
sensitive to freezing temperatures. Consequently, colder temperatures are likely to cause N. fowleri to
encyst in lake and river sediment, which offers more protection from cold and freezing water
temperatures.
Source: http://www.cdc.gov/parasites/naegleria/pathogen.html
How does the brain-eating ameba cause disease?
Answer: The ameba infects humans when contaminated water enters the body through the nose. It
then travels up the olfactory nerve to the brain, where it causes PAM. Contaminated water can come
from different types of untreated water sources.
What is primary amebic meningoencephalitis (PAM)?
Answer: Answers will vary. PAM is an acute disease that rapidly leads to death in approximately all
cases. Only 3 of 133 U.S. patients have survived the infection. On average, humans die <10 days after
exposure to the ameba. The disease presents much like bacterial meningitis, which can lead to delayed
diagnosis and treatment. In approximately 25% of cases, the trophozoites were observed in the
cerebrospinal fluid while the patient was still alive, but more often, the diagnosis is not made until
after the patient dies and an autopsy is performed. In those cases, the ameba is usually observable in
tissue sections of the brain.
18
Diagnostics and treatment strategies
What are some diagnostic strategies for brain-eating amebic infection and PAM?
Answer: Answers will vary. Diagnostic strategies include (1) direct visualization (motile trophozoites
observed in the cerebrospinal fluid), (2) immunohistochemistry (antibody staining of the amebae in
tissue), (3) polymerase chain reaction testing (detecting ameba DNA), and (4) culture (amebae grown
by using bacteria as a food source and incubating at high temperature).
What are the treatment options for brain-eating ameba infection/PAM?
Answer: Treatment includes a combination of antibiotics and an investigational drug (miltefosine).
The documented survival rate when using the combination or cocktail of antibiotics is 2% (3 of 133
cases). Miltefosine has been used in other ameba infections and appears to improve survival rates.
Surveillance and epidemiology
How many cases of PAM were reported during 1962–2014?
Answer: The number of cases of PAM reported during 1962–2014 was 133, with a range of 0–8
infections/year.
Who is most likely to be infected with this brain-eating ameba?
Answer: Male children. The median age of patients reported is 11 years, and 77% were male.
When is the most common time of the year to become infected with this brain-eating ameba?
Answer: The most common months of illness onset are July and August, with a range of April–
November.
Where is exposure to the brain-eating ameba most likely to occur?
Answer: The most probable water exposure sources are lakes, ponds, and reservoirs (untreated fresh
water). Texas and Florida have the highest number of reported cases in the United States.
Environmental prevention challenges
What are certain environmental concerns regarding N. fowleri infection?
Answer: A major concern is that as water temperatures increase worldwide, the geographic range of
brain-eating ameba is anticipated to shift northward. Another concern is moderate chlorine resistance,
which affects water treatment as a prevention measure.
19
Appendix 3A
Waterborne Disease Case Study
Laboratory Exploration
Name: __________________________________ Date: ________________
Directions: In groups of 3–4, complete the case study laboratory exploration.
Assume that the samples provided are from the northeastern Arizona lake where the boy with brain-
eating ameba was swimming. Pretend you are a team of microbiologists. Each team will receive a
sample to observe. With your team, create wet-mount slides of your water sample. Use the microscopes
to explore organisms in your sample. In the spaces below, draw the organisms you see. Be sure to
include the magnification level. Next, to identify the scientific name of the organism, use the Pond Life
Identification Kit at http://www.microscopy-uk.org.uk/index.html?http://www.microscopy-
uk.org.uk/pond/index.html and http://www.microscopy-uk.org.uk/index.html?http://www.microscopy-
uk.org.uk/pond/protozoa.html.
Table 1: Organisms in your sample
Picture:
Organism Group: _________________________
Magnification: _________________________
Picture:
Organism Group: _________________________
Magnification: _________________________
Picture:
Organism Group: _________________________
Magnification: _________________________
Picture:
Organism Group: _________________________
Magnification: _________________________
20
Picture:
Organism: _________________________
Magnification: _________________________
Picture:
Organism: _________________________
Magnification: _________________________
Picture:
Organism Group: _________________________
Magnification: _________________________
Picture:
Organism Group: _________________________
Magnification: _________________________
Picture:
Organism Group: _________________________
Magnification: _________________________
Picture:
Organism Group: _________________________
Magnification: _________________________
21
Questions
1. What was the most common organism found?
2. Approximately, how many amebae were in your sample?
3. If N. fowleri was not found in the lake samples in the case of the Arizona male, it might still have
been possible for infection to be caused by N. fowleri.
a. Explain why the male was diagnosed with primary amebic meningoencephalitis likely caused by
N. fowleri.
b. Why was swimming in the lake the likely cause of N. fowleri infection?
4. If N. fowleri were present in the lake samples in the case of the Arizona boy, explain why more
people were not infected. Hint: consider the organism’s life cycle and influencing environmental
factors (e.g., climate, water pH, nutrient availability, and spatiality).
5. Explain why CDC does not recommend testing rivers and lakes for N. fowleri.
6. N. fowleri infections are rare. During 2005–2014, a total of 35 infections were reported in the United
States. Of those cases, 31 persons were infected by contaminated recreational water. What are some
more likely dangers related to swimming in recreational water, such as a lake?
22
Appendix 3B
Waterborne Disease Case Study
Laboratory Exploration Answer Key
Name: __________________________________ Date: ________________
Directions: In groups of 34, complete the case study laboratory exploration.
Assume that the samples provided are from the northeastern Arizona lake where the boy with brain-
eating ameba was swimming. Pretend you are a team of microbiologists. Each team will receive a
sample to observe. With your team, create wet-mount slides of your water sample. Use the microscopes
to explore organisms in your sample. In the spaces below, draw the organisms you see. Be sure to
include the magnification level. Next, to identify the scientific name of the organism, use the Pond Life
Identification Kit at http://www.microscopy-uk.org.uk/index.html?http://www.microscopy-
uk.org.uk/pond/index.html and http://www.microscopy-uk.org.uk/index.html?http://www.microscopy-
uk.org.uk/pond/protozoa.html.
Table 1: Organisms in your sample
Picture:
Organism Group: _________________________
Magnification: _________________________
Picture:
Organism Group: _________________________
Magnification: _________________________
Picture:
Organism Group: _________________________
Magnification: _________________________
Picture:
Organism Group: _________________________
Magnification: _________________________
23
Picture:
Organism: _________________________
Magnification: _________________________
Picture:
Organism: _________________________
Magnification: _________________________
Picture:
Organism Group: _________________________
Magnification: _________________________
Picture:
Organism Group: _________________________
Magnification: _________________________
Picture:
Organism Group: _________________________
Magnification: _________________________
Picture:
Organism Group: _________________________
Magnification: _________________________
24
Questions
1. What was the most common organism found?
Answer: Answers will vary.
2. Approximately, how many amebae were in your sample?
Answer: Answers will vary.
3. If N. fowleri was not found in the lake samples in the case of the Arizona male, it might still have
been possible for infection to be caused by N. fowleri.
a. Explain why the male was diagnosed with primary amebic meningoencephalitis likely caused by
N. fowleri.
Answer: The laboratory test results from the spinal fluid confirmed that the male was infected with
N. fowleri. He presented with early symptoms of primary amebic meningoencephalitis, such as
fever and headache, within the average time of symptom onset. He later presented with neck
stiffness.
b. Why was swimming in the lake the likely cause of N. fowleri infection?
Answer: Answers can include: Swimming in warm freshwater lakes is the most common type of
exposure that leads to the brain-eating ameba infection. However, the water temperature on that
day, September 8, 2007, was 86.3°F (30.2°C), which was not within the optimal water temperature
range for the growth of N. fowleri (45ºC–55ºC).
Note: Consider discussing what epidemiologists might do at this stage. For example,
epidemiologists would systematically identify other cases in the area and look at the distribution
(e.g., person, place, and time) and possible determinants of infection (i.e., other sources of
infection, including bodies of warm freshwater, such as lakes and rivers; geothermal water,
including hot springs; warm water discharge from industrial plants; geothermal drinking water
sources; swimming pools that are poorly maintained, minimally chlorinated or unchlorinated; water
heaters; and soil) across cases.
4. If N. fowleri were present in the lake samples in the case of the Arizona boy, explain why more
people were not infected. Hint: consider the organism’s life cycle and influencing environmental
factors (e.g., climate, water pH, nutrient availability, and spatiality).
Answer: Multiple factors are needed that must come together at the same time to result in infection.
N. fowleri infects person when water containing the ameba enters the body through the nose, and N.
fowleri must be in an infective life-cycle stage. Life-cycle stages are influenced by environmental
factors that affect an organism’s ability to feed, grow, and survive. Humans will not be infected if
the water ingested does not contain the ameba, if life cycle conditions are not ideal for infection, or if
the water is ingested other than through the nose.
5. Explain why CDC does not recommend testing rivers and lakes for N. fowleri.
Answer: The ameba is naturally occurring and no established relationship between detection or
concentration of N. fowleri and risk for infection is known.
25
6. N. fowleri infections are rare. During 2005–2014, a total of 35 infections were reported in the United
States. Of those cases, 31 persons were infected by contaminated recreational water. What are some
more likely dangers related to swimming in recreational water, such as a lake?
Answer: Drowning; infection (e.g., leptospirosis, swimmers itch [cercarial dermatitis], giardiasis,
cryptosporidiosis, cholera, shigellosis, norovirus, and Escherichia coli infections, including E. coli
O157:H7 infections); wound infections (e.g., the flesh-eating bacteria Aeromonas); diving hazards
and other water-related injuries. (Answers will vary.)
26
Appendix 4
Waterborne Disease Case Study
Public Awareness Campaign
Name: __________________________________ Date: ________________
Directions: Create a public awareness campaign through a most-wantedtype poster or a brochure that
illustrates how to reduce the risk for acquiring a particular waterborne disease.
1. Choose and circle a recreational water illness from the following
Crypto” (Cryptosporidium)
Giardia
Hot tub rash
Legionella
Swimmer's ear (otitis externa)
Head lice
Methicillin-resistant Staphylococcus aureus
Pinworm
2. Complete the graphic organizer below by using the CDC Internet site concerning Recreational Water
Illnesses and Prevention Tips: http://www.cdc.gov/healthywater/swimming/rwi/illnesses/index.html.
What is it?
Why should you be
concerned about it?
How is it spread in
aquatic facilities?
How do you protect
yourself and your
family?
3. Use your graphic organizer to create a public awareness campaign through a most-wantedtype
poster or a brochure that illustrates how to reduce the risk for acquiring a particular waterborne
disease.
27
Appendix 5A
Brain-Eating Ameba Quiz
Name: __________________________________ Date: ________________
Directions: Circle the best answer for each question.
1. The scientific name of the brain-eating ameba is . . .
a. Haemophilus influenzae.
b. Escherichia coli.
c. Naegleria fowleri.
d. Salmonella typhi.
2. From the case study, this pathogen is detected in . . .
a. skin.
b. hair.
c. bones.
d. cerebrospinal fluid.
3. Which of the following is NOT a morphologic stage of this pathogen’s life cycle?
a. Cyst
b. Polyp (Answer)
c. Trophozoite
d. Flagellate
4. The optimal temperature range for the growth of this pathogen is . . .
a. 45ºC –55ºC.
b. 10ºC –20ºC.
c. 25ºC –45ºC.
d. 75ºC –90ºC.
5. Which of the following can reduce your risk for becoming infected with PAM?
a. Avoid swimming in lakes and ponds during periods of high water temperature.
b. Hold your nose shut, use nose clips, or keep your head above water when taking part in water-
related activities in bodies of warm fresh water.
c. Avoid disrupting the sediment (digging or stirring) while taking part in water-related activities in
shallow, warm freshwater areas.
d. All of the above.
28
Appendix 5B
Brain-Eating Ameba Quiz Answer Key
Name: __________________________________ Date: ________________
Directions: Circle the best answer for each question.
1. The scientific name of the brain-eating ameba is . . .
e. Haemophilus influenzae.
f. Escherichia coli.
g. Naegleria fowleri. (Answer)
h. Salmonella typhi.
2. From the case study, this pathogen is detected in . . .
e. skin.
f. hair.
g. bones.
h. cerebrospinal fluid. (Answer)
3. Which of the following is NOT a morphologic stage of this pathogen’s life cycle?
e. Cyst
f. Polyp (Answer)
g. Trophozoite
h. Flagellate
4. The optimal temperature range for the growth of this pathogen is . . .
e. 45ºC –55ºC. (Answer)
f. 10ºC –20ºC.
g. 25ºC –45ºC.
h. 75ºC –90ºC.
5. Which of the following can reduce your risk for becoming infected with PAM?
e. Avoid swimming in lakes and ponds during periods of high water temperature.
f. Hold your nose shut, use nose clips, or keep your head above water when taking part in water-
related activities in bodies of warm fresh water.
g. Avoid disrupting the sediment (digging or stirring) while taking part in water-related activities in
shallow, warm freshwater areas.
h. All of the above. (Answer)