UWM Libraries Research and Course Guides: WInSTEP Environmental Health Science Program: Zebrafish

Student Resources

Click on the following links for more handy resources:

Zebrafish as Models: Studying the Effects of Environmental Agents on Human Health

Module: Zebrafish as Models

The goal of this module is for students to conduct scientific investigations using the zebrafish to understand how environmental agents affect human development. By applying the results of the zebrafish embryo experiments to human embryo development, students draw conclusions regarding personal health, environmental hazards, and the risks and benefits of personal and social decisions in relation to these hazards.

Introduction to Zebrafish
This video was created by Dr. Michael Carvan from the UW-Milwaukee School of Freshwater Sciences and the UWM WInSTEP SEPA Program to explain the important role of zebrafish in research.
Zebrafish Module Teacher Workshop 1
This is the first of two training workshop videos for teachers for the zebrafish module.
Zebrafish Module Teacher Workshop 2
This is the second of two teacher training workshop videos for the zebrafish module.
Serial Dilutions

Zebrafish Embryo Diagrams - Control
Parent/Guardian Letter for the Zebrafish Module
This letter provides a description of the zebrafish module and the ethical use of animals in the project.
Safety Data Sheet - Caffeine
Safety Data Sheet - Ethanol
Safety Data Sheet - Nicotine

Zebrafish - Background Information

Key literature resources and website links about zebrafish in general:

All You Need to Know About Acclimation
This article discusses the benefits of acclimation and the most commonly-used methods used to acclimatize fish and other inhabitants. The article then provides a detailed step-by-step set of instructions for the Drip Method, the method that is generally accepted among aquarists to be the best-practice acclimation method.
AZ Animals: Zebrafish
Provides background information about zebrafish including general facts, classification, etc.
Dissection of Organs from the Adult Zebrafish
A Golden Fish Reveals Pigmentation Loss in Europeans
Human skin color, determined by the pigment melanin, is a highly variable trait. The reason for these variations has been an enduring mystery of biology. Although sunlight exposure contributes to skin pigmentation, much of the variation we see is likely genetic. To better understand the genetic origin of skin color variation in humans, the authors turned to the zebrafish (Danio rerio), a popular model organism, which—like humans—also displays variations in skin color.
Sex Determination of Zebrafish
Your Inner Fish
Our arms, legs, necks and lungs were bequeathed to us by a fish that lumbered onto land some 375 million years ago. The genetic legacy of this creature can be seen today in our own DNA, including the genes used to build our hands and limbs.
Zebrafish in the Classroom
Zebrafish in the Classroom is designed to be a resource for teachers and students who are using zebrafish in the classroom. The site includes common techniques for spawning and raising zebrafish, experiments and virtual experiments, games, tutorials, and a glossary.
Zebrafish Journal
Zebrafish is the only peer-reviewed journal that focuses on solely on zebrafish. This link is to the special issue: Zebrafish in Education, and includes information about zebrafish programs, student authored articles, curricula, and abstracts.
Zebrafish - A Key to Understanding Human Development
This NIH video shows how the zebrafish, Danio rerio, provides a valuable resource for scientists trying to understand the intricate process by which a fertilized egg develops into a fully formed individual—and also the numerous diseases and conditions that can result when even a tiny part of that process goes wrong. The video includes interviews with NIH researchers and footage of the NIH zebrafish facility. Thought to be the largest in the world, the facility has enough space for 19,000 tanks to accommodate 100,000 fish.
Zebrafish 10 Facts

Go Fish! Developmental Stages in Zebrafish (Poster)

Zebrafish Embryo Development

Key literature resources and website links related to zebrafish embryo development:

The Art of Breeding Laboratory Zebrafish
This presentation provides an in-depth look at our current understanding of zebrafish reproduction, as well as a comprehensive overview of some of the most critical factors impacting the breeding performance of this important laboratory animal.
Classroom Explorations: Zebrafish Development Videos and Images
Images of Danio rerio (zebrafish) Embryonic Development
Stages of Embryonic Development of the Zebrafish
Ultraviolet Irradiation Impairs Epiboly via Microtubules in Zebrafish Embryos
Zebrafish Atlas (Virtual Slide Library)
Zebrafish Development
Includes detailed images, movies, and details for each stage of development.
Zebrafish Developmental Movies: 2-cell stage to 16-somite stage
Zebrafish Embryo Development - 24 hours in 46 seconds
Zebrafish Embryo Development Flipbook
This series of images depicts the first 48 hours of zebrafish development that can be made into a flipbook.
Zebrafish Made to Grow Pre-hands instead of Fins
Killifish Can Adjust Their Egg-laying Habits in Response to Predators, Study Shows
Killifish can change their egg-laying habits in response to predators in the area in order to survive.

Toxicants and Zebrafish Embryo Development

The following images show what happens to the developing brain of a zebrafish embryo exposed to nicotine:

Adverse Effects in the Fish Embryo Acute Toxicity (FET) Test: A Catalogue of Unspecific Morphological Changes Versus More Specific Effects in Zebrafsh (Danio rerio) Embryos
Provides an overview of both common and selected more specific morphological effects seen in zebrafsh embryos after exposure to a wide variety of chemical substances.
Fish Reveal Caffeine and Other Common Drugs May Alter Human Facial Development
Impact of substances on zebrafish embryos shows how the prenatal development of human facial features might also be affected.
Movement Disorder and Neuromuscular Change in Zebrafish Embryos After Exposure to Caffeine
Ethanol Effects on the Developing Zebrafish: Neurobehavior and Skeletal Morphogenesis
The Effects of Ethanol on Zebrafish Development
Effects of Ethanol on Photoreceptors and Visual Function in Developing Zebrafish
Embryonic Nicotine Exposure Disrupts Adult Social Behavior and Craniofacial Development in Zebrafish
Nicotine Response Genetics in the Zebrafish
Glyphosate Induces Neurotoxicity in Zebrafish
Glyphosate based herbicides (GBH) like Roundup® are used extensively in agriculture as well as in urban and rural settings as a broad spectrum herbicide. Its mechanism of action was thought to be specific only to plants and thus considered safe and non-toxic. However, mounting evidence suggests that GBHs may not be as safe as once thought.
Control of Developmental Speed in Zebrafish Embryos Using Different Incubation Temperatures
The zebrafish is a valuable model organism that is widely used in studies of vertebrate development. In the laboratory, zebrafish embryonic development is normally carried out at 28.5°C. In this study, researchers sought to determine whether it was possible to modify the speed of embryonic development through the use of short- and long-term variations in incubation temperature.

Acute and Chronic Toxicity of Nitrate to Early Life Stages of Zebrafish-Setting Nitrate Safety Levels for Zebrafish Rearing
Recirculating aquaculture systems (RAS) have been widely used for zebrafish rearing, allowing holding of many thousands of fish at high densities. Water quality in RAS largely depends on biofilters that ultimately convert the extremely toxic ammonia excreted by fish into the much less toxic nitrate. However, when water renewal is minimal in RAS, nitrate can accumulate to high enough levels to negatively impact fish welfare and performance.
Zebrafish Models in Neural and Behavioral Toxicology across the Life Stages
The article discusses the value of using zebrafish in neural and behavioral toxicology research.

Zebrafish Microscopy Archive Presentation

Nicotine, Healthy Development, and Zebrafish

The following informational presentation was created and copyrighted by Dr. Kurt Svoboda, Associate Professor in the Laboratory for Developmental Neurobiology in the University of Wisconsin-Milwaukee Joseph J. Zilber School of Public Health.

Zebrafish Microscopy Archive Videos

The following informational videos were created and copyrighted by Dr. Kurt Svoboda, Associate Professor in the Laboratory for Developmental Neurobiology in the University of Wisconsin-Milwaukee Joseph J. Zilber School of Public Health. These videos correspond with the presentation, "Nicotine, Healthy Development, and Zebrafish" in the LibGuide box above.

Toxicants and Human Development

Key literature resources and website links related to toxicants and human embryo development:

Lethal Doses of Water, Caffeine and Alcohol
Describes the lethal doses for water, caffeine and alcohol.
Centers for Disease Control and Prevention: Fetal Alcohol Spectrum Disorders
Addressing Fetal Alcohol Spectrum Disorders (Substance Abuse and Mental Health Services Administration)
How Blood Alcohol Concentration Impairs Body and Behavior
Blood alcohol concentration (BAC) is a measurement of the percent of alcohol is in a person's bloodstream. This, rather than the exact amount of alcohol the person has consumed, will determine the effects the alcohol will have on the person's brain, body and behavior. There are many factors that affect BAC, including size, gender, physical condition; what has been eaten and when; how much sleep is logged; medications; and alcohol content of drinks.
Prenatal Exposure to Alcohol Disrupts Brain Circuitry: No Safe Level of Drinking During Pregnancy, Neuroscientist Says
Prenatal exposure to alcohol severely disrupts major features of brain development that potentially lead to increased anxiety and poor motor function, according to neuroscientists at the University of California, Riverside.
Vital Signs: Alcohol-Exposed Pregnancies, U.S. 2011-2013
Alcohol is a teratogen. Prenatal alcohol exposure is associated with a range of adverse reproductive outcomes and can cause fetal alcohol spectrum disorders (FASDs) characterized by lifelong physical, behavioral, and intellectual disabilities. FASDs are completely preventable if a woman does not drink alcohol while pregnant.
Air Pollution, Fetal and Infant Tobacco Smoke Exposure, and Wheezing in Preschool Children: A Population-based Prospective Birth Cohort
This study examines the associations of exposure to air pollution with the risk of wheezing in preschool children, and assessed whether these are affected by tobacco smoke exposure.
Early Data in E-Cigarette Study May Raise Safety Concerns
A laboratory study presented early this year reported that the nicotine-laced vapor generated by an electronic cigarette promoted the development of cancer in certain types of human cells much in the same way that tobacco smoke does.
Effects of Nicotine During Pregnancy: Human and Experimental Evidence
Evaluation of the Association between Maternal Smoking, Childhood Obesity, and Metabolic Disorders: A National Toxicology Program Workshop Review
This review looks at the association between childhood obesity and metabolic disorders as they relate to maternal smoking duirng pregnancy as part of a broader effort to develop a research agenda to better understand the role of environmental chemicals as potential risk factors.
Fetal and Neonatal Nicotine Exposure and Postnatal glucose homeostasis: Identifying Critical Windows of Exposure
Flavoring Chemicals in E-Cigarettes: Diacetyl, 2,3-Pentanedione, and Acetoin in a Sample of 51 Products, Including Fruit-, Candy-, and Cocktail-Flavored E-Cigarettes
A Hot Debate Over E-Cigarettes as a Path to Tobacco, or From It
This article in The New York Times debates the public health implications of e-cigarettes.
‘I Can’t Stop’: Schools Struggle With Vaping Explosion
E-cigarettes have been touted by their makers and some public health experts as devices to help adult smokers kick the habit. But school officials, struggling to control an explosion of vaping among high school and middle school students across the country, fear that the devices are creating a new generation of nicotine addicts.
Know the Risks: E-Cigaretts and Young People
E-cigarette use poses a significant – and avoidable – health risk to young people in the United States. Besides increasing the possibility of addiction and long-term harm to brain development and respiratory health, e-cigarette use is associated with the use of other tobacco products that can do even more damage to the body. Even breathing e-cigarette aerosol that someone else has exhaled poses potential health risks.
More Teens Now Try Vaping Than Smoking
Trying conventional cigarettes is no longer a teen rite of passage, but trying e-cigarettes might be becoming one, a new government survey suggests. Nearly half of high school students surveyed in 2015 tried vaping, while only a third smoked a tobacco cigarette, according to the nationwide survey of 15,000 students released by the federal Centers for Disease Control and Prevention.
Nicotine's Binding Mystery Solved
Caltech chemists have solved the chemical mystery of why nicotine binds to acetylcholine receptors with high affinity in the brain but with low affinity in muscles. Such information will help researchers understand nicotine addiction and guide drug discovery efforts for diseases such as schizophrenia and Parkinson's disease.
Nicotine’s Effect on the Developing Embryo Studied in New Model
Selling a Poison by the Barrel: Liquid Nicotine for E-Cigarettes
This article discusses the dangers of liquid nicotine for e-cigarettes. The liquid nicotine, along with various flavorings, colorings, and other chemicals, are powerful neurotoxins and can cause vomiting, seizures, and can be lethal.
Smoking in Pregnancy Tied to Kids' Conduct Problems
New study shows that children of women who smoked are more likely to have behavioral problems that those whose mothers didn't smoke.
Teens Using Vaping Devices in Record Numbers
America’s teens report a dramatic increase in their use of vaping devices in just a single year, with 37.3 percent of 12th graders reporting “any vaping” in the past 12 months, compared to just 27.8 percent in 2017. These findings come from the 2018 Monitoring the Future (MTF) survey of a nationally representative sample of eighth, 10th and 12th graders in schools nationwide, funded by a government grant to the University of Michigan, Ann Arbor.
Tobacco Smoke Affects Early Human Embryonic Development
U.S. Chamber of Commerce Works Globally to Fight Antismoking Measures
Vaping 'Pumps' Cancer-Causing Substances into the Lungs
Many studies conducted over the past couple of years have found that, despite being advertised as a safer alternative to traditional cigarettes, electronic cigarettes (e-cigarettes) actually hide numerous threats to health.
Is Caffeine Consumption Safe During Pregnancy?
Caffeine Improves Memory in Alzheimer's Mice
This article highlights new research that shows for the first time that caffeine has a positive effect on tau deposits in Alzheimer's disease.
Caffeine in Pregnancy (March of Dimes)
Caffeine Shakes Up Growing Mouse Brains
When pregnant rodents consumed this stimulant, offspring had altered neurons and faulty memory.
No Coffee for Pregnant Moms?
A new study in mice suggests that at larger doses, caffeine can influence fetal brain development and increase the risk of having seizures.
Trends in Caffeine Intake Among US Children and Adolescents
EPA Issues Emergency Order to Stop Use of Pesticide Dacthal to Address Serious Health Risk
The U.S. EPA announced an emergency suspension of all use of the pesticide dimethyl tetrachloroterephthalate (DCPA or Dacthal). The EPA has taken this action because unborn babies whose pregnant mothers are exposed to DCPA could experience changes to fetal thyroid hormone levels, which has been linked to low birth weight, impaired brain development, decreased IQ, and impaired motor skills later in life, some of which may be irreversible.
Little Things Matter: The Impact of Toxins on PreTerm Birth
Babies who are born preterm -- which is 3 or more weeks before their due date -- are more likely to die in the first year of life or develop heart disease, diabetes or have trouble learning. Every year about 15 million babies are born too soon. Preterm birth, which is a key barometer for the health of a population, can be due to infection, poor nutrition, or having twins or triplets, but these only account for about one-third of babies born preterm. This video is about how widespread exposure of pregnant women to toxic chemicals can “tip the scales” and increase the number of babies born too soon.
Little Things Matter: The Impact of Toxins on the Developing Brain
UCSF Presentation Reveals Glyphosate Contamination in People Across America
Glyphosate, the most used herbicide in the world, has been found in the urine of 93% of the American public during a unique testing project that started in 2015. Glyphosate, labeled a ‘probable human carcinogen’ by the World Health Organization’s cancer agency IARC in 2015, has now been revealed to be ubiquitous in the first ever comprehensive and validated LC/MS/MS testing project to be carried out across America.
The Womb is No Protection From Toxic Chemicals
Until a few decades ago, the popular but falsely reassuring belief was that babies in the womb were perfectly protected by the placenta and that children were just “little adults,” requiring no special protections from environmental threats. We now know that a host of chemicals, pollutants and viruses readily travel across the placenta from mother to fetus, pre-polluting or pre-infecting a baby even before birth.

E-Cigarettes

Zebrafish Module E-Cigarette Extension: Aerosol Collection Demonstration
This video provides a demonstration of how to collect the aerosol for the e-cigarette experiment.
Case of Severe Lung Illness
This engaging module takes you through a video case study to witness the real-life experiences related to vaping. As you move through this case study, you will read through information, and analyze tables, videos and data to be able to answer the case study questions.
Early Data in E-Cigarette Study May Raise Safety Concerns
A laboratory study presented early this year reported that the nicotine-laced vapor generated by an electronic cigarette promoted the development of cancer in certain types of human cells much in the same way that tobacco smoke does.
E-Cigarettes and Lung Health-American Lung Association
Facts about health risks and e-cigarettes.
E-cigarettes, "Vapes" and JUULs: What Teens Should Know
Flavoring Chemicals in E-Cigarettes: Diacetyl, 2,3-Pentanedione, and Acetoin in a Sample of 51 Products, Including Fruit-, Candy-, and Cocktail-Flavored E-Cigarettes
There are > 7,000 e-cigarette flavors currently marketed. Flavoring chemicals gained notoriety in the early 2000s when inhalation exposure of the flavoring chemical diacetyl was found to be associated with a disease that became known as “popcorn lung.”
How Mitch McConnell Accidentally Created An Unregulated THC Market
‘I Can’t Stop’: Schools Struggle With Vaping Explosion
Johns Hopkins Researchers Find Thousands Of Unknown Chemicals In Electronic Cigarettes: Study Identifies Compounds Undisclosed by Popular Brands Including Industrial Chemicals, Caffeine
Vaping aerosols contain thousands of unknown chemicals and substances not disclosed by manufacturers, including industrial chemicals and caffeine, Johns Hopkins University researchers found.
Know the Risks: E-cigarettes and Young People
E-cigarette use poses a significant – and avoidable – health risk to young people in the United States. Besides increasing the possibility of addiction and long-term harm to brain development and respiratory health, e-cigarette use is associated with the use of other tobacco products that can do even more damage to the body. Even breathing e-cigarette aerosol that someone else has exhaled poses potential health risks.
Lung Damage From Vaping Resembles Chemical Burns, Report Says
Doctors at the Mayo Clinic examined samples of lung tissue from 17 patients, all of which looked as if the people had been exposed to toxic chemicals, the researchers said.
More Teens Now Try Vaping than Smoking
Teens Using Vaping Devices in Record Numbers
America’s teens report a dramatic increase in their use of vaping devices in just a single year, with 37.3 percent of 12th graders reporting “any vaping” in the past 12 months, compared to just 27.8 percent in 2017. These findings come from the 2018 Monitoring the Future (MTF) survey of a nationally representative sample of eighth, 10th and 12th graders in schools nationwide.
Teen Vaping is on the Rise, and Their Favorite Flavor is Flying Under the Radar
The number of teen e-cigarette users in the United States is still on the rise, according to a new demographic study—and most of what they're inhaling is mint-flavored.
Vaping 'Pumps' Cancer-causing Substances into the Lungs
Many studies conducted over the past couple of years have found that, despite being advertised as a safer alternative to traditional cigarettes, electronic cigarettes (e-cigarettes) actually hide numerous threats to health.
The Vape Talk - American Lung Association
Over 5,700 kids start vaping every day. Misconceptions, peer pressure and marketing tactics all contribute to the staggering rise. This conversation guide provides information for the discussion.
'We need to tell everyone': Teens, young adults who vape more at risk for COVID-19, study says
A new study has found that vaping is linked to an elevated risk of COVID-19 among teenagers and young adults, providing more evidence of the harmful effects of electronic cigarettes.

Electronic Cigarette Toxicology Research Using Zebrafish Embryos as a Developmental Model (Teacher's Guide)
Calculating the Amount of Vape Needed for the Experiment
This spreadsheet shows the calculations for making the stock solution from the vape, as well as each of the dilutions.
E-cigarette PowerPoint Presentation
This presentation provides background on the anatomy of an e-cigarette, the toxicity of the chemicals in e-cigarettes, and how the aerosol affects the lungs.
Pathology of Vaping-Associated Lung Injury

WInSTEP Environmental Health Science Program: Zebrafish