Bronxville High School Page 1 of 8 Environmental Science Blackworm Toxicity Lab Mrs. Bastone/Mr.

Ippolito
Name: Claire McGonigle
INTRODUCTION
In this experiment, we will determine the behavior of California black worms after being exposed to different concentrations of alcohol. The black worms natural behavior consists of the worms clumping together in ball. If this does not happen after being exposed to the alcohol, this would be an indicator that the alcohol had an effect on the worms. These worms live in muddy sediments, especially in shallow water along the edges of marshes and ponds throughout the United States and many other parts of the world. They do not have a skeleton but the fluid inside them gives them support and form. When touched, the worms will attempt to escape in a circular motion or by reversing its body. When the alcohol is added with the worms, it will enter the worms system through ingestion and will attack the worms digestive track and blood stream. The alcohol could also enter the worm through absorption through the skin and also be carried in to the blood stream and other parts of the body.

HYPOTHESIS: HYPOTHESIS:
When alcohol is added into the water that the California black worms are in, the worms will be affected by absorption and ingestion and will not behave as they normally do.

PROCEDURE
First, we will obtain 8 weigh boats and label them Control, WControl, Low, Medium, High, Water1, Water2, and Water3. Then we will fill the weigh boats labeled WControl, Water1, Water2, Water3 with 40 mL of distilled water and fill a 100 mL beaker with 60 mL of the alcohol solution. With the 50 mL of distilled water and pipettes we will make Solution #1(low alcohol concentration) by adding 1 mL of the alcohol and 39 mL of distilled water and pour it into the weigh boat labeled low. To make solution #2 we will mix 10 mL of alcohol and 30 mL of distilled water and pour it into the weigh boat labeled medium. For solution #3 we will pour 40 mL of alcohol in to the weigh boat labeled high. By using a pipette or spoon we will transfer 40-50 worms into the remaining distilled water in the 500 mL beaker. Then we will transfer about 10 of those worms to the WControl, 10 to the Water1, 10 to the Water2 and 10 to Water3. After this, we will observe the worms behavior and will monitor how they would normally act. Then we will transfer the worms to the weigh boats labeled low, medium and high and start timing and record behavior observations after 3, 6, and 10 minutes. After the 10 minutes are up, we will transfer the worms back into the weigh boats labeled WControl, Water1, Water2, and Water3, this will be the recovery time for the worms. Again, we will observe the behavior at 3, 6, and 10 minutes. After this is done, we will record our data and begin to clean up. The picture below demonstrates how the lab will look with the weigh boats and 500 mL with worms.

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Bronxville High School Page 2 of 8 Environmental Science Blackworm Toxicity Lab Mrs. Bastone/Mr. Ippolito
DATA/OBSERVATIONS DATA/OBSERVATIONS TA/

Swimming Behavior 0 minutes Notes: 3 minutes Notes: Control 2 Moving normally 3 Swimming around a lot 3 Still swimming around 3

Alcohol Toxicity Data Low Conc 1 Little movement 1 Little movement 1 Still very little movement 1 Medium Conc 0 No movement 0 All died High Conc 0 No movement 0 All started bleeding and died 0 Dead

6 minutes Notes:

0 Dead

Still active Barely moving Dead Dead Rate Activity as 0 (Not Active) 1 2(normally active) 3 4 (very active)

10 minutes Notes: Swimming Behavior

Caffeine Toxicity Data Control Low Conc 1 Barely moving 1 Still barely moving 2 Started to behave normally 2 2 Swimming normally 2 Swimming normally 2 Swimming normally 2
Continued to swim normally

0 minutes Notes: 3 minutes Notes:

6 minutes Notes:

Medium Conc 1 Barely moving 1 Still barely moving 1 Barely moving 2

High Conc 1 Barely moving 0 No movement

Continued Began to swim Still little normally normally movement behavior Rate Activity as 0 (Not Active) 1 2(normally active) 3 4 (very active)

10 minutes Notes:

1 Started little movement 1

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Bronxville High School Page 3 of 8 Environmental Science Blackworm Toxicity Lab Mrs. Bastone/Mr. Ippolito
Swimming Behavior 3 minutes Notes: Control R Swimming around normally R Fully recovered R Fully recovered Alcohol Toxicity Recovery Low Conc P Little to no movement P Little movement P Still not swimming as normal Medium Conc N;dead High Conc N;dead

6 minutes Notes: 10 minutes Notes:

N;dead

N;dead

N;dead

N;dead

Rate Activity as R (complete recovery) P (partial recovery) N (no recovery)

Swimming Behavior Control 3 minutes Notes: R Fully recovered

Caffeine Toxicity Recovery Low Conc P Little movement, one dead after transfer N No movement N No movement Medium Conc P Little movement High Conc R Completely recovered

6 minutes Notes: 10 minutes Notes:

R Swimming normally R Continuing normal behavior

R Swimming normally R Continuing normal behavior

R Swimming normally R Continuing normal behavior

Rate Activity as R (complete recovery) P (partial recovery) N (no recovery)

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Bronxville High School Page 4 of 8 Environmental Science Blackworm Toxicity Lab Mrs. Bastone/Mr. Ippolito
Time 0 min 3 min 6 min 10 min 0 min 3 min 6 min 10 min 0 min 3 min 6 min 10 min 0 min 3 min 6 min 10 min Gr 1 Class Data For Alcohol Gr 2 Gr 3 Gr 4 2 2 4 3 2 3 3 2 3 3 3 3 1 2 3 1 1 2 1 1 2 1 0 2 0 1 1 0 0 1 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Class Data For Caffeine Gr 2 Gr 3 Gr 4 2 4 2 4 2 3 2 2 1 2 1 3 2 3 2 1 1 1 1 2 1 2 2 1 1 3 0 1 1 2 1 2 Gr 5 2 2 2 2 2 2 2 2 1 1 0 0 0 0 0 0 Gr 6 4 3 2 2 4 2 2 0 3 1 0 0 0 0 0 0 Ave 2.8 2.5 2.3 2.1 2.5 1.6 1.6 1.16 1.3 .6 .3 .16 .16 0 0 0

Control

Low

Medium

High

Control

Low

Medium

High

Time 0 min 3 min 6 min 10 min 0 min 3 min 6 min 10 min 0 min 3 min 6 min 10 min 0 min 3 min 6 min 10 min

Gr 1 2 2 2 1 2 3 3 3 3 3 4 4 1 3 1 2

Gr 5 4 4 4 4 4 3 3 3 4 3 2 2 4 1 0 1

Gr 6 3 3 2 2 3 2 2 2 0 1 1 1 1 0 0 0

Ave 3 2.8 2.5 2 2.5 2.5 2.5 2.2 2.4 2 2 2 1.8 1 .6 1

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Bronxville High School Page 5 of 8 Environmental Science Blackworm Toxicity Lab Mrs. Bastone/Mr. Ippolito
Exposure Bar Graph for Alcohol Activity Rating 4 3 2 1 0 minutes 0 3 6 10 0 3 6 10 0 3 6 10 0 3 6 10 Control Low Medium High Exposure Bar Graph for Caffeine Activity Rating 4 3 2 1 0 minutes 0 3 6 10 0 3 6 10 0 3 6 10 0 3 6 10 Control Low Medium High
RESULTS
Answer all of the questions from the handout here.

1. In this experiment, exposure frequency would refer to how many times the worms were exposed to the toxicant. Exposure duration would refer to the amount of time the worms stayed in the toxicant (10 minutes). Lastly, exposure concentration would refer to the concentration of the toxicants in the distilled water( low, medium, high).

2. Acute testing was done in this investigation because the worms were exposed to high concentrations of a toxicant over a short period of time(10 minutes) as opposed to chronic testing which would be low concentrations of exposure over a long period of time. The benefit of using acute testing would be that the exposure frequency would be a lot lower than during chronic testing. The benefit of using a chronic toxicity test would be that there would be a lower concentration of the toxicant than in an acute testing.

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Bronxville High School Page 6 of 8 Environmental Science Blackworm Toxicity Lab Mrs. Bastone/Mr. Ippolito
3. A chronic toxicity test that we might perform on the black worms would be to lower the concentrations that were used in this investigation and test/observe the black worms for about a year and check on their behavior every day for about an hour. My hypothesis would be that the black worms would eventually have the same affects to the toxicants as they did in this lab but it would just drag out over a longer period of time.

4. The exposure pathway for our toxicant would be through ingestion and absorption. When the alcohol was added with the worms, it entered the worms system through ingestion and attacked the worms digestive track and blood stream. The alcohol also entered the worm through absorption through the skin and carried in to the blood stream and other parts of the body.

5. Age would affect the results of our toxicants because the young worms and the old worms would be more affected than the middle-age worms due to weaker immune systems . Genetic difference would also affect the results because worms that were born with diseases would also be more sustainable to be affected more than a worm that was born healthy. Body size also affects the results because the smaller bodies can not take as high of a dosage of a toxicant as larger bodies can so they would be more affected than the bigger worms.

6. A sublethal concentration of the toxicant would be a concentration of a potentially lethal substance but the concentration is not high enough for it to be lethal. A lethal concentration is the concentration of a substance that is high enough to cause death.

7. The body systems in the worms that were involved in the detoxification process would be the blood stream and the digestive/immune system. At the low concentration of alcohol, the toxification was reversible. For both the medium and high concentrations of alcohol it was irreversible because all the worms were already dead. For caffeine, both the medium and high concentrations were reversible but the low concentration was not.

8. N/A

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Bronxville High School Page 7 of 8 Environmental Science Blackworm Toxicity Lab Mrs. Bastone/Mr. Ippolito
9. At each concentration, the worms showed different behavior, not all the worms acted the same way. This could be explained by the age, genetic differences, and body sizes of the worms. A young worm would act different from a middleaged worm. A genetically healthy worm would act differently from a non-healthy worm and a smaller worm would act differently than a larger worm.

10. The control in this experiment would be the pure water because it was not changed in any way. A control is necessary in an actual scientific experiment because the controls give a baseline against which to compare experimental results.

11. Pesticides, insecticides, and herbicides would all harm the environment and in a chain reaction, would affect the worms as well.

12. Lifestyles affect the health, metabolism, and limits of exposure to toxins of individuals. Risk increases if a person is in poor health. Limits of exposure to toxins can be affected by action such as frequency of a toxin. Ricks increases as the frequency increases.

13. No, not necessarly. Although humans and vertebrates contain an immune/digestive system and blood stream like the worms, there would be a different affect. For example, if this test was performed on humans, the concentration levels of the toxins would have to be a lot higher due to the fact that humans are a lot bigger than the black worms and we could handle a much larger concentration without it being lethal.

14. (See conclusion below)

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Bronxville High School Page 8 of 8 Environmental Science Blackworm Toxicity Lab Mrs. Bastone/Mr. Ippolito
CONCLUSION
In this experiment, we determined the behavior of California black worms in different concentrations (low, medium, high) of caffeine and alcohol. One of the main indicators that the black worms are behaving normally is when they clump in a ball. However, this did not occur in any of the concentrations. Once the worms were placed in the medium and high concentrations of the alcohol, they were barely swimming around and they all died in less than 3 minutes. The worms in the low concentration of alcohol survived although they were not moving around as much as they normally would. However, once placed back in the distilled water for recovery, they completely recovered after 10 minutes and were behaving normally again. The worms that were placed in the low, medium and high concentrations of caffeine for 10 minutes were barely moving. For a while we actually thought they were dead until we started touching them and they began to move around again. During the recovery period, the worms in the medium and high concentrations were completely detoxified but the detoxification of the low concentration of worms was irreversible. This being an acute toxicity test, and the exposure frequency and duration was low but had a high exposure concentration, I think it was too much for the worms and they were not able to behave normally due to to absorption and ingestion of the toxins. In the end, my hypothesis that when the toxins were added to the water and then the worms were added as well, the worms were affected by the toxins greatly through absorption and ingestion and were unable to pertain to their normal behavior.

REFLECTION
Overall, I think that this lab was successful. It was an enjoyable and interesting experiment. Also, this lab was very understandable. In this lab, we learned how the black worms would react when placed in toxins. When placed in caffeine, we learned that the black worms were still active but were not reacting as they normally would. We also discovered that some of the black worms were harmed when being transported from the control to the different concentrations. When placed in medium and high levels of alcohol, we learned that the black worms just died. When in low levels, the black worms barely survived. This was an informative lab and it was successful.

End of Lab Report Form