The Tell-tale Heart of Daphnia: Environmental Spices

WARM-UP

1. Turn in homework1. Turn in Lab Notebook at the end of the period

2. Put signed papers in PER 1-2 INBOX

2. Take out a scratch sheet of paper and solve the following

3. List out things you consider a toxin. What is not a toxin?

Paracelsus, 1493 - 1541

“The right dose differentiates a poison and a remedy”

Anything can be a toxin!

160 pounds

What is LD50?

LD 50 (Lethal Dose 50)is the dosage or amount of a chemical that causes 50% of the population react (usually die). For Example: An experiment testing tobacco gives

10 people ½ a teaspoon of tobacco. How many people should die if the LD50 is ½ a teaspoon?

How would you find the LD50 of a chemical?

Toxicologists find the LD50 by performing a test called dose-response assessments where they expose model organisms to varying concentrations of a chemical.

Another way to measure response to toxins…

Daphnia heat rate test

What kind of toxins can we find in the water?

http://www.colbertnation.com/the-colbert-report-videos/232640/july-01-2009/nicholas-kristof

Figure adapted from Edwards et al. 2006

Trends in wildlife reproductive health

Figure adapted from Edwards et al. 2006

Trends in wildlife reproductive health

Figure adapted from Edwards et al. 2006

Trends in wildlife reproductive health

Figure adapted from Edwards et al. 2006

Trends in wildlife reproductive health

Sharpe & Irvine 2004

Trends in human reproductive health

EDCsEndocrine disrupting compounds

Incidences of altered reproductive health grouped by geographic area, suggesting an environmental cause

Endocrine disrupting compounds (EDCs)- alter hormonal and homeostatic systems

Endocrine system

Diamanti-Kandarakis et al. 2009

System of communication between tissues via chemical signals (e.g. hormones) that regulates:

Development

Growth

Metabolism

Reproduction

Calcium regulation

Immune function

Glucose homeostasis

Digestion

Circadian rhythm

Formation of adipose tissue

EDCs- chemical intruders

How can chemicals alter reproductive health?

Endocrine system uses chemical signals (hormones) - how tissues “talk” to each other

Endocrine system critical for gonad development and reproductive function

Internal signal

brain gonad

How can chemicals alter reproductive health?

Endocrine system uses chemical signals (hormones) - how tissues “talk” to each other

Endocrine system critical for gonad development and reproductive function

EDCs- chemical intruders

External signal

Limited knowledge about EDCs

How can we identify which chemicals act like hormones?

What are the effects?

What happens when we’re exposed to low-dose mixtures?

How can we calculate risk?

Where are these compounds found in the environment?

1996 Congress passed legislation to create the EPA Endocrine Disruptor Screening Program to assess potential risk to human health and wildlife

endocrine disruptionliver lesions

cancer

endocrine disruptionincreased risk of disease

damaged nervous system

Health effects on fishes from urban areas

Human diseases linked to EDCs

Diamanti-Kandarakis et al. 2009

Development

Growth

Metabolism

Reproduction

Calcium regulation

Immune function

Glucose homeostasis

Digestion

Circadian rhythm

Formation of adipose tissue

Limited knowledge about EDCs

How can we identify which chemicals act like hormones?

What are the effects?

What happens when we’re exposed to low-dose mixtures?

How can we calculate risk?

Where are these compounds found in the environment?

Spices - nautral & human-made chemicals from foods we eat or plants, e.g. vanilla

Pharmaceuticals - human-made chemicals manufactured for use as medicinal drugs, e.g. ibuprofen

Plasticizers - human-made chemicals added to plastics to make them flexible, soft, etc., e.g. dibutyl phthalate (DI-BE-OO-TAHL THAL-ATE)

Industrial solvents - chemicals that are good at dissolving substances, natural or human-made, e.g. benzaldehyde (BENS-AL-DUH-HIDE)

Perfumes - a natural or human-made chemicals used to create a nice smell or mask odors, e.g. linalool (LIN-AH-LEW-AL)

Lilial

EDCs in common household items

Dibutyl phthalate

Bisphenol A

Ibuprofen

Spices - nautral & human-made chemicals from foods we eat or plants, e.g. vanilla

Pharmaceuticals - human-made chemicals manufactured for use as medicinal drugs, e.g. ibuprofen

Plasticizers - human-made chemicals added to plastics to make them flexible, soft, etc., e.g. dibutyl phthalate (DI-BE-OO-TAHL THAL-ATE)

Industrial solvents - chemicals that are good at dissolving substances, natural or human-made, e.g. benzaldehyde (BENS-AL-DUH-HIDE)

Perfumes - a natural or human-made chemicals used to create a nice smell or mask odors, e.g. linalool (LIN-AH-LEW-AL)

Cinnamon (Cinnamic Acid)

Spices Treated in Wastewater

Spices Treated in Wastewater

Veratraldehyde (waffle cone smell)

Spices Treated in Wastewater

Natural vanilla

Spices Treated in Wastewater

Artificial vanilla

A natural spice

Vanilla

$63.92/pint

Or a man made flavoring

$4.38/pint

Industrial Solvents

Your Mission

1. Choose a freshwater water source you are interested in and perform a basic water testing on the source you collected

2. Choose a toxin you are interested in experimenting with Types of toxins I have readily available

(http://www.pesticideinfo.org/List_AquireAll.jsp?Rec_Id=PC42993&Taxa_Group=Fish) Caffeine Theobromine (chocolate) Thymol (thyme) Cinnamic Acid Cinnamaldehyde Ethyl Vanillin Vanillin Ibuprofen Detergent and Oil

3. Test the toxin on Daphnia to find the LD50 and a heart rate response

4. Lab Report of Findings, Earth Day and Letter to State Representative

What to do

Get into groups of 2-3

Think and research about where you want to get water samples from and what toxin you are interested in testing and why.

.Research already known effects of toxin and save it on a google doc or any file. You will need it later. Keep a list of sources too!!

Tell Quanina when you have decided.

www.pesticideinfo.org

Toxins I Have

Caffeine Theobromine

(chocolate) Thymol (thyme) Cinnamic Acid Cinnamaldehyde Ethyl Vanillin Vanillin Ibuprofen Detergent and Oil

Before We Begin

Brushing up on metric conversions, dilutions, and pipetting

Unit Conversion Ladder Method BS Method Algebraic Method

Pipetting How to pipette A test of your pipetting skills

Concentrations and Dilutions ?

Writing your Methods

Needs to be in paragraph form

Can split up the 3 experiments Water testing LD 50 or dose-response assessment test Heart-rate test

Water testing and collecting, be sure to include the latitude and longitude of where you got the water. http://itouchmap.com/latlong. html How much water you got The depth Average temperature What you observed about the water For actual testing you can write see aqua pharmaceutical master fresh

water testing kit for methods on water testing

Methods must include…

LD 50 or Dose Response Assessment Test: How much water you will pour into each jar Your concentration of toxin in each jar How many daphnia you put in each jar and how you will transfer the

daphnia How you will record your quantitative (#) and qualitative (words) data Demo how to transfer Daphnia

Heart rate test How you will place the daphnia under the microscope The concentration of the toxin you will be using How many daphnia you will be using How you will record your quantitative (#) and qualitative (words) data Demo how to take heart rate test

EMAIL METHODS BY END OF PERIOD