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IBL WORKSHOPS ZOOS MUSEUMS PARKS SCHOOLS

 Water Quality Assessments Using Ceriodaphnia Dubia

An inquiry-based lesson plan designed to promote critical thinking by

integrating content with traditional and inquiry-based learning.

All Hands-On Learning is NOT Inquiry-Based Learning

Inquiry based learning is process-oriented and does not focus on a single correct answer, but
rather emphasizes the process of gathering information and forming a conclusion. Traditional
hands-on learning tends to be product-oriented and has students follow a pre-planned procedure
to come to a single, specified answer.

1
 Key Terms
Process skills (PS)
Skills that students will engage in while thinking critically. These include observing,
questioning, predicting, planning, investigating, interpreting, and communicating. These
skills are found in each step of the inquiry process.
Investigating Surroundings
Observing the overall surroundings. What do I see? What is understood about the topic?
What still needs to be understood?
Narrowing Focus
Observing student needs and interests, as well as academic content. Find the balance
between natural curiosity and standards-based concepts. What area can be concentrated
on to best promote growth and learning?
Questioning
Forming questions about what is not fully understood. Comparative questions can be
investigated. They need to be able to lead into an action plan. What can be found out?
Uncover Prediction
Logically thinking to form a prediction about what could happen. What do I expect to
happen based on my experiences and knowledge?
Initiate Plan
Figure out the action plan. Design an experiment which will answer the comparative
question. What can I do to answer this question? How can I find this out?
Research and Collect Data
Investigating the elements of the experiment. Researching and collecting data that applies
to the action plan.
Examine Results
Interpreting the data collected. What does this data mean? What else do I want to find
out?
Communicating
Communicating the information that was found to someone else. The way the data is
presented. What will the audience want to know? What will the audience be able to
understand about this?

2
 Table of Contents
1. Overview and Practical Application 4
2. Lesson Plan 5
a. Objectives
b. Educational Standards
c. Materials
d. Vocabulary
3. Introduction 7
4. Instructional Overview 8
5. Post-Assessment 8
6. Journaling 9
7. Background Science 10
8. About Inquiry-Based Learning as it Applies to this Lesson Plan 12
9. Pre-Experiment Preparation 14
10. Experiment: Group Procedures 16
11. Appendix 1 – Data Sheet 17
12. Appendix 2 – Vocabulary Defined 18
13. Appendix 3 – Student Feedback Form 19
14. Appendix 4 – Teacher Feedback Form 20
15. Appendix 5 – Experiment Calendar 21
16. Appendix 6 – Photos 22

3
 Overview
This is an inquiry-based learning lesson in which students learn about ​C. dubia​ and their habitat.
Students will be engaged in hands-on activities to culture ​C. dubia​ and set-up an experiment to
answer a comparative question regarding reproduction under various environmental conditions.
Finally, the project will link IBL with the biology involved in evolution and how species adapt.
More questions will emerge to continue scientific investigations and further the learning process.

Practical Application
This experiment allows students to understand toxicity levels in their own water samples, using
techniques that would be used in a real lab setting. Each student will have practice with the
scientific process, including interpreting and documenting data.

4
 Lesson Plan
Grade Level: 7-9

Class:

Teacher:

Time Required/Duration: Day one (Friday)- 40 minutes/ full class period

Each day- take 10 minutes to feed and count the water fleas
Final day (Next Friday)- 40 minutes/ full class period

Objectives
1. Incorporate the process of inquiry-based learning into a traditional direct instruction
classroom setting.
2. Assess the toxicity of various surface waters from different environmental settings.
3. Students will be able to identify the life cycle, morphology, and habitat of ​C. dubia​.
4. Students will recognize ways that species, specifically ​C. dubia​., adapt to their
environment.
5. Differences in adaptation will lead to a comparative question that the teacher will provide
to the class.
6. Students will work together in groups to maintain stock cultures of ​C. dubia​ and
complete an action plan to explore various adaptations to different environments.
7. Students will collect, tabulate and present scientific data as well as journal their process.
8. Students will uncover additional questions and think critically about the data found.

Educational Standards
Scientific inquiry​ – Students develop scientific habits of mind as they use the processes of
scientific inquiry to ask valid questions, and to gather and analyze information. They understand
how to develop hypotheses and make predictions. They can reflect on scientific practices as they
develop plans of action to create and evaluate a variety of conclusions. Students are also able to
demonstrate the ability to communicate their findings to others.

Life Science (Topic: Species and Reproduction)

8.LS.1​ Diversity of species, a result of variation of traits, occurs through the process of
evolution and extinction over many generations. The fossil records provide evidence that
changes have occurred in number and types of species.
8.LS.2 ​ Every organism alive today comes from a long line of ancestors who reproduced
successfully every generation.
8.LS.3​ The characteristics of an organism are a result of inherited traits received from
parent(s).

5
 Materials
Needed​:
1. Stereoscope (magnifying glass will work, but it won't be as clear)
2. Four different sources of naturally occurring water (i.e., river, pond, lake,
stream, ditch, etc.)​1

IBL Institute Provided at No Charge (Shipping and handling fees will apply)​:
1. C. dubia​ stock cultures in moderately hard synthetic rearing water
2. Laboratory supplied culture water to transfer organisms
3. Eye Droppers
4. Sampling cups with lids
5. Disposable 30 ml cups
6. C. dubia​ food supply: YCT (yeast, Cerophyll and trout chow) and
unicellular green algae (​Selenastrum capricornutum)​ .
7. Cheesecloth to filter samples

Nice to Have Items (Grant funding may be available)​:

1. Grow light bulb (no fixture) - estimated cost $15 to $20 2​
2. Digital or mechanical timer – estimated cost $12 to $30
3. Lighted magnifying glass – estimated cost $20-$50

To order the IBL Institute provided supplies, please contact Jessica Begonia at 419-223-1362.
Supplies will be scheduled for delivery two days before the experiment starts.

Vocabulary
(See Appendix 4 for Vocabulary Defined)

C. dubia/ Daphnia
Asexual reproduction
Sexual reproduction
Brood
Neonates

1
​There is an opportunity here to allow the students to inquire even more about the environment around them. If
possible, instruct your students to bring the water samples from home. When the results vary, it will make the
student question the environment they live in which is the essence of inquiry learning.
2
​There are other grow light bulbs with fixtures that can be used as a substitute. If not accessible, setting cultures
near a window will work as well.

6
 Introduction
The teacher will:
1. Show the photo of the daphnia on the board and briefly discuss the morphology of
C.dubia.​ (See Appendix 6, page 22)
2. Lead a discussion of the ecology, life history, and life cycle of ​C. dubia.​
(​INVESTIGATE SURROUNDINGS)
3. Explain that students will be taking home a sample cup and bringing back a water
sample to use in the experiment. Approve of each student’s location for the water
sample, to ensure differences in the samples. ​(NARROW FOCUS)
a. Sample sources could include ponds, ditches, streams, aquariums, culture
water from the sampling kits, etc.
4. Provide information to bridge the scientific background with the hands-on part of
the lesson.
5. Review the difference between sexual and asexual reproduction.
6. Explain that the lesson will use asexually reproducing organisms that are
genetically identical (clones).
7. Discuss how species can adapt to their environment and evolve.
8. Let students know they will be working in groups of 4 to 5 dependent on class size.
9. As a class, come up with a comparative question. ​(ASK COMPARATIVE
QUESTIONS)
a. For example, will average brood sizes as measured by the total number of
young in three broods be larger in more nutrient rich waters (like agricultural
runoff)?
b. Which student thinks water from around their house will produce the largest
broods? Why/Why not?

7
 Instructional Overview
1. Introduce the students to the materials provided, discuss the logistics and dynamics of the
hands-on learning and make assignments.​3
2. Students will observe the daphnia (both in the water through a microscope and through
photography) and sketch one. 4​
3. They will proceed with the actual experiment.
4. Students will collect and record data.
5. Provide the detailed background information to the students with a lecture.
6. The teacher may play the video provided regarding C.dubia and acute toxicity if time
allows.

Post-Assessment 5​
1. After the experiment is over and the data has been recorded, post the comparative
question again for students to read.
2. Ask students to think about their answer to this question and the processes that lead them
to get to that answer.
3. Students will be given a partner from a different group​6​. They will write to that partner:
a. The steps they took to find the answer.
b. Why those steps led them to the conclusion they have.
c. What is left unanswered, or any new questions they have after the experiment.
4. After writing, have them share their paragraph with their partner to read.
5. While reading their partner’s work, students should think about:
a. What is different about their partner’s work?
b. What caused it to be different from their own work?
c. Does the way their partner thought about the process change the way they
personally think about it?
6. Have the partners discuss their thinking with one another. ​(COMMUNICATE
FINDINGS)
7. If students have changed their thinking in any way from what they had written, they can
make notes on what has changed and what made them change their mind.
8. Students will turn in their writing to the teacher. Check to make sure:
a. They have stated their conclusion to the question.
b. Their conclusion was made logically using the data found in the experiment

3
T​his is what is covered in the anticipatory set. This part could take up to two class periods.
4
​Students can infer what part of the daphnia is which and what purpose each part serves.
5
​This can be done using pencil and paper or using the preferred technology of the classroom.
6
If groups get different answers, try to pair students from groups with different answers to the question. This will
create more points of discussion and can expand their thinking on their own answer​.

8
 Journaling
Students will document their data, thoughts, and questions every day for the duration of this
lesson. This can be done with technology such as Google Docs or Word, with pencil and paper in
a notebook, or any other method that best fits the class. Each class, the students will respond to
prompts provided by the teacher. For class periods covering the anticipatory set, students will
respond to prompts at the end of class discussing the material, as well as any questions they
have. For class periods spent on the experiment, while students are working they will record data
and respond to prompts discussing what they observe.

Introduction prompts​:
What made you excited about today’s lesson?
What is one thing that you learned today?
What is one thing you are curious about?

Experiment prompts​:
What differences did you observe?
- Size, color, movements
- Draw a picture of the differences, if possible

What have you noticed about the ​c. dubia​ today, and what do you think that means?
- If they died, why do you think that happened?
- What could you do now?
- What could you learn using the materials available to you?

Experiment data​:
How many ​C. dubia​ are in each container?
A
B
C
D

9
 Background Science
Life Cycle
● C. dubia​ are filter feeders and feed on algae and other particulates through their movement.
● During the normal part of their life cycle they reproduce asexually. Show the following
video:
○ https://www.youtube.com/watch?v=i9zj9V8OWRk
○ Diploid eggs develop into larvae that grow in a brood chamber.
○ Young are typically released after about three days.
● Throughout their life cycle ​C. dubia​ adapt to extreme environmental conditions including
over population, cold weather, lack of food, and toxic environments.

Reproduction
● Under extreme environmental conditions ​C. dubia​ reproduce sexually and fertilized eggs
can remain dormant for extended periods of time until environmental conditions improve.
○ This is important because sexual reproduction allows for evolution of the species
maintaining some sense of gene flow. ​Daphnia​ mutate to adapt to environmental
conditions and then pass genes on to future generations. This combination helps the
species to survive.
○ When the ​C. dubia​ recognizes changes in the environment, she will give birth to
males (instead of just females) so that sexual reproduction can occur.
○ An asexually reproducing female ​C. dubia​ can have three broods in as little as six
days.
○ Because of this short life cycle, mutations can occur quickly, and the species can
adapt easily.
○ Daphnia​ can easily provide insight into evolution because they reproduce quickly,
they are easily observed, and adaptations are often quick and in direct response to
their environment.
■ One adaptation to extreme environmental conditions is the ability to
reproduce sexually.

Adaptation
● A number of genetic traits in ​C. dubia​ are evident and observable. These traits include
mobility, size, age, morphology, behavior, predator response, and their immune system.
● Predators can also bring about traits that result in adaptations.
○ For example, when predatory fish are around, daphnia will respond with smaller
offspring; conversely in the presence of midges(small flies), they will often have
larger offspring.
○ Kairomones are the chemicals present in the predators that induce changes in the
Daphnia​ that result in a change in the size of offspring
● Daphnia​ have the ability to adapt to a broad range of environmental conditions; this is
known as ecological generalization. Narrow environmental tolerance is referred to as
ecological specialization. ​Daphnia​ can adapt to increasing levels of salinity.
○ Two challenges for an organism in a saline environment are dealing with osmosis
and toxicity from the sodium and chloride. Usually, generalization results in some
cost, but ​daphnia​ populations that have a broad range of tolerance do better with
increasing levels of salinity, than do populations with narrow environmental

10
 tolerance. There does not seem to be an obvious cost to the ​daphnia​ that lends
itself to a remarkable ability to survive and evolve.
○ C. dubia​ are one of the species used to assess toxicity of water. If we put some ​C.
dubia​ into pond water, and they die, that water is said to be acutely toxic. If they
live, but reproduce at lower than expected rate, the water is said to exhibit chronic
toxicity.

Distribution
● Daphnia​ are one of the most studied species. It is an important species because it is
widespread and important to the ecology. This zooplankton spans over much of the
northern hemisphere, and was thought to have spread as a result of glacier activity. The
genus is regularly found in some regions (including Japan) but not to others (like Russia)
and so it is difficult to pinpoint the exact area of speciation.

11
 About Inquiry-Based Learning As It Applies To This Lesson Plan

This is a project that works best when students work in small groups (4-5). Inquiry is
collaborative in nature. The process takes advantage of students’ strengths to contribute to the
project. Some are great communicators, some are problem-solvers, and some have
well-developed technical skills. In the workplace, we also work in groups. We work as part of a
team. The inquiry process develops skills necessary to solve complex problems in the world.

The students will benefit from background science of ​C. dubia.​ Make sure the section on the
background science is available to each group. Some in the group may have little interest in the
material provided whereas others will want to read it in depth, but the end result will be that
everyone in the group will know more about the test organisms. It’s important for them to know
that ​C. dubia​ are indigenous to most streams, rivers, ponds, and many other natural surface water
sources. Connect the relevance of the group working with this test species as it relates to the
comparative question. ​C. dubia​ are one of the species used to assess toxicity of water. If we put
some ​C. dubia​ into pond water and they die, that water is said to be acutely toxic. If they live but
reproduce at lower than expected rate, the water is said to exhibit chronic toxicity. Students will
either bring in water samples from a local pond, stormwater outfall, stream, standing water in the
yard, etc. or the samples would be provided by you. Have enough source water for each group to
work with two of the water sources. Four water sources would be fine for 5 or 6 groups.

Investigate your surroundings and narrow your focus:​ Encourage each group to observe the
test organisms. Have them get familiar with the light and magnifier to see how the ​C. dubia
move. Have them transfer some test organisms using the dropper to new water. It is important
that they transfer organisms carefully to avoid introducing air bubbles, which may damage the
organism. Have them discuss different sizes, what they see, how they move, and what techniques
help them see the organisms best. This is a good time for students to write down questions. We
suggest having them write individual questions on individual pieces of paper so the questions can
later be sorted. It’s not important to sort those now, but this can be revisited once the students
are more in tune with the inquiry process.

Ask comparative questions:​ At the heart of inquiry is the comparative question. Comparative
questions are ones that can be investigated. Some questions are very good questions, but they
are very difficult to investigate. For example: Why are butterflies attracted to my flower garden?
Good question, but difficult to investigate. However, we can take that question and change it to:
Are butterflies attracted more to red flowers or white flowers? Do you see where we are going
with this? You can now design an experiment to count how many butterflies visited each of the
colors and compare the results. What will come of this will be more questions. Do the findings
hold up for different kinds of butterflies? Another experiment can be designed. In the case of
the ​C. dubia,​ the comparative question, at least initially, is being provided by you. This makes
the lesson plan a guided inquiry. An open inquiry is one in which the students pick the topic,
create the questions, create the action plan, etc. The comparative question for this lesson plan is:
Does water from source A exhibit more or less toxicity than from source B? Later, we will
revisit the questions the students asked above and have them separate those questions that can be
investigated and those that cannot. ​Often, questions that would be difficult to investigate can be
made investigable by turning them into comparative questions​.

12
Uncover your prediction:​ We are not talking about group-think here. What do you
individually think? At this point, each group has two water samples from different sources and a
supply of C​. dubia,​ and soon they will have an experiment to follow. Each student will have a
prediction and they should record that prediction. Which water sample will exhibit more
toxicity? Will it be the water from the aquarium in the room, or will it be from a local pond?
Some observation is involved here. Are there differences in turbidity? Color? Debris? A
prediction is not the same thing as a hypothesis. A hypothesis might be: All swans are white. A
prediction would be: I think the next swan I see will be white. A prediction is based upon the
individual’s experiences, observations, opinions, knowledge, and instincts.

Initiate an action plan: ​ The experiment has been provided. The action plan was designed to
help students answer the comparative question. Neither you nor I know which water sources will
exhibit more or less toxicity. It’s not one of those “do the experiment and I will let you know the
right answer.” There is no right answer. Ask the students to record their data on the data sheet
provided. To make future lesson plans or repeats of this one more inquiry-based, simply ask
them to make a data sheet and record their findings. Perhaps have half the groups use the data
sheet provided and then let the other half come up with their own. Part of inquiry requires you to
give up some control to allow your students to figure it out.

Examine results and communicate findings:​ Each group will present their findings to the
class. Typically, each group would prepare a graph, data table, chart, pictures or whatever they
want to communicate their findings to the class. A poster or section of white board is helpful.
The group goes to the front and each member usually participates. As a group, they share which
water samples they used and any noteworthy observations. They share their individual
predictions. They share the mortality of the test species, the reproduction rate, and their findings
regarding toxicity. How do they answer the comparative question? What did they learn from the
experience? Classmates then have an opportunity to ask questions to the presenting group.

Student/Teacher Roles for Each Step:

Investigate your surroundings: The students are doing this

Narrow your focus: The teacher and students are doing this
Ask comparative question: The teacher is doing this
Uncover your prediction: Each student is doing this
Initiate an action plan: The teacher is doing this
Research and data collection: The students are doing this*
Examine results and communicate findings: The students are doing this
*The teacher or students may generate the data sheet

13
 Pre-Experiment Preparation
Below are the steps to be taken by the teacher/students in preparation for the start of the
experiment.

Wednesday​: In your kit, you will find the YCT and unicellular green algae, along with the
daphnia stock culture and eye dropper. Use the eyedropper to place six or seven drops of each
food source in the provided mass culture.

Now that the organisms are fed, you need to provide them with a day and night cycle. In the lab,
we use a grow light on a timer to maintain a constant 16 hours of daylight and 8 hours of night.
You can do the same, or you can simply put your cultures near a window.

Before the start of the experiment, you will need to have your students divided into 4 groups of
4-5 students. Either you or your students will need to collect 4 samples of naturally occurring
water. The samples can be collected from a pond, a stream, or a ditch. Daphnia particularly like
aquarium water, so if you have an aquarium in your class or at home, you can also use that. You
can also use the culture water provided in your sampling kit. We have provided sample bottles to
use for collection. It is good technique to document collection information on the bottle and
subsequent data sheets used. Note the date and time, who collected the sample, where it is from,
and any other pertinent information.

Thursday: ​Take the provided laboratory water and gently shake to aerate it. It is a good idea to
gently shake the culture water and all future samples prior to transferring the organisms to
provide some air for the daphnia. Never use chlorinated water as this will kill the daphnia.
Pour about 60 mL of the fresh culture water provided in your lesson plan kit into a separate,
small bowl.

Next, add about 6-7 drops of the food sources to the fresh culture water. Finally, you will need to
capture some of the larger daphnia and transfer them to the fresh culture water. The daphnia can
be viewed best with some light from below and a magnifying glass. Identify the adults and
transfer approximately 10 of them using the provided eye dropper. Please note, this eye dropper
is specially prepared with a little wider hole to ensure the test organisms are not damaged while
handling them. The adults separated today will produce the neonates that we will draw upon for
the start of the test on Friday.

Be sure to feed both the new culture and the mass culture 6-7 drops of the food supplied and
return them to their place near a window or timed grow light.

14
Friday Morning: ​Separate out an adult and 4-5 neonates for each group using the culture started
Thursday. If you have four groups, you will need 16 total neonates. To distribute the samples to
your student groups, you can either opt to have each group have one water source to test, or you
can have each group test all 4 water sources by sharing the 4 source samples between the groups.

If possible, put a drop of food into each cup Saturday and/or Sunday.

15
 Experiment: Group Procedures
Friday: ​Take one of the cups with the adult daphnia and her brood. The mom will be the large
one and the babies will be very small. Observe under bright light and magnification and sketch
the mom on the bottom of the data sheet. 7​

Write down the comparative question that your teacher provided. What do you (not the group,
but you) think the answer to the comparative question will be? ​(UNCOVER PREDICTION)

Take 4 of the small plastic cups and label them with your group number and “A”, “B” “C” and
“D.” Gently shake the sample of your various water sources and add the appropriate sample to
each cup, filling about ¾ of the way. Using an eyedropper, add one drop of food. Next, using
the eyedropper, bright light, and a magnifying glass if needed, transfer a neonate to each cup.

Place your 4 cups under the fluorescent light with the timer set to 16 hours on (daylight) and 8
hours off (nighttime), or near the window​8​. We will be counting the total number of
young/neonates produced over the next week from these babies (soon to become moms). Predict
how many will be in the cup on Monday, when you return. 9​

Note: We are now finished with the original cup containing the mom and her brood.

Monday: ​By Monday or Tuesday, the​ ​neonates should have matured and produced broods of
their own. Prepare four more sample cups by gently shaking the samples and pouring 20 mL of
sample into each cup, and adding one drop of each food source to the new sample cups. Transfer
the adult daphnia from each sample cup to the corresponding fresh sample leaving the neonates
behind. ​(RESEARCHING AND COLLECTING DATA)

Count the neonates and record the data for each cup using the data sheet provided. Place your 4
cups under the fluorescent light with the timer set to 16 hours on (daylight) and 8 hours off (night
time), or near the window. Rinse out the four cups from which you transferred daphnia using
distilled water so you can reuse the provided cups on alternating days.

Tuesday: ​Add one drop of food to each cup.

Wednesday: ​Repeat the procedures from Monday.

Thursday: ​Add one drop of food to each cup.

Friday: ​Repeat the procedures from Monday and Wednesday. The original daphnia that started
as neonates at the beginning of the experiment will have had 3 broods. Tabulate your data from
the week and plan on how you will present your findings. ​(EXAMINE RESULTS AND
COMMUNICATE FINDINGS)

7
Ask if the mother and her brood are identical. How do the students know this? The mom is reproducing asexually.
The babies are all “clones.” That is, they are genetically identical.
8
Be sure to place it close enough to the window that it receives light, but not too close that it gets cold overnight.
9
Have the students predict how many babies will be in the cup tomorrow and ask them what information leads to
this prediction.

16
 Appendix 1 – Data Sheet

Group # ____
Student Names:__________________________________
__________________________________
__________________________________
__________________________________

Sample water “A” : __________________________________

Sample water “B” : __________________________________
Sample water “C” : __________________________________
Sample water “D” : __________________________________

Number of “babies” in cup

A B C D

Mon ( / ) _______ _______ _______ _______

Wed ( / ) _______ _______ _______ _______

Fri ( / ) _______ _______ _______ _______

TOTAL: _______ _______ _______ _______

17
 Appendix 2 – Vocabulary Defined

C. dubia/ Daphnia:​ Commonly known as “water fleas”, these small creatures are filter feeders
who reproduce asexually, but under certain conditions they can reproduce sexually.

Asexual reproduction:​ Reproduction that only requires one female individual. All of the
offspring will be genetically identical to the mother, or “clones” of the mother.

Sexual reproduction:​ Reproduction that requires two individuals, a male and a female, who
combine their genetic material to create offspring. The offspring will be genetically different
from their parents and siblings.

Brood:​ The group of young produced at one time (siblings)

Neonates:​ Newborn offspring

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 Appendix 3 – Student Feedback

1. Did the initial ​daphnia​ in all 4 cups with different waters have the same
number of babies?

2. If you repeated the test using babies from Monday, would you expect
similar results, or might some adapt to the environment and do better?

3. What did you learn from this?

4. What additional questions come to mind after having done the experiment?

5. Was it fun and/or interesting?

19
 Appendix 4 – Teacher Feedback

1. What evidence suggests students grasped the major themes of the experiment
(i.e., life cycle of ​C. dubia,​ culturing techniques, reproduction of ​C. dubia,​
IBL, evolution, etc.)?

2. Do you anticipate other guided or open inquiry projects arising from this
project? What questions did the students ask that suggest understanding
and interest in the subject?

3. To what extent did this project fit into your curriculum and teaching
agenda?

4. Would you consider doing this again?

5. What would improve the experience?

Please return completed surveys to ​[email protected]

20
 Appendix 5 – Experiment Calendar

S​UNDAY M​ONDAY T​UESDAY W​EDNESDAY T​HURSDAY F​RIDAY S​ATURDAY

1 2

3 4 5 6 7 8 9

TRANSFER START TEST and Optional: FEED

ADULTS INTO FEED
CULTURE CUPS
and FEED
10 11 12 13 14 15 16
Optional: FEED TRANSFER FEED TRANSFER FEED COUNT YOUNG
ADULT TO NEW ADULT TO NEW and END TEST
TEST WATER TEST WATER
Optional: Optional:
and FEED. and FEED.
TRANSFER TRANSFER
COUNT YOUNG. COUNT YOUNG.
ADULT TO NEW ADULT TO NEW
TEST WATER. TEST WATER.
COUNT YOUNG. COUNT YOUNG.

17 18 19 20 21 22 23

24 25 26 27 28 29 30

31

21
Appendix 6 – ​Ceriodaphnia​ d​ubia​ Photos

22
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