ap® investigation #12 - ap biology...

AP® InvestIgAtIon #12 Ecology: BEhavior – TEachEr’s guidE

©2012, Ward’s Natural Science

All Rights Reserved, Printed in the U.S.A.

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250-7460 v.5/12

Kit # 3674-12

aBsTracT 1

gEnEral ovErviEw 1

rEcording daTa 2

MaTErial rEquirEMEnTs/chEcklisT 4

naTional sciEncE EducaTion conTEnT sTandards 5

corrElaTion To aP conTEnT sTandards 5

TiME rEquirEMEnTs 5

lEarning oBjEcTivEs 6

safETy PrEcauTions 7

PrE-laB PrEParaTions 8

Background 11

ParT 1: cEll sizE & diffusion 13

ParT 2: ModEling osMosis in living cElls 17

ParT 3: osMosis in living PlanT cElls 21

assEssMEnT quEsTions/addiTional quEsTions (oPTional) 24

furThEr inquiry invEsTigaTions 23

TEachEr’s answEr kEy 24

table of Contents

**AP® and the Advanced Placement Program are registered trademarks of the College Entrance Examination Board. The labs and materials in this kit were developed and prepared by WARD’S Natural Science Establishment, which bears sole responsibility for their contents..

WILL FIX ALL PAGE #’S ONCE EVERYTHING ELSE IS FINALIZED.



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Ecology: BEhavior – TEachEr’s guidE Kit # 3674-12

AbstrACtOrganisms orient to stimuli that are important to their survival. Movement toward or away from important stimuli (taxis) depends upon both the sensory and motor abilities of the organism. This lab explores the chemotactic behaviors that fruit flies and/or pill bugs exhibit when exposed to the controlled environment of a choice chamber. Students identify patterns in the behaviors and make inferences based on the composition of the tested materials and the organisms’ responses. Students then determine what materials and experimental paradigms will be tested further.



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generAl overvIewThe College Board has revised the AP Biology curriculum to begin implementation in the fall of 2012. Advanced Placement (AP) is a registered trademark of the College Entrance Examination Board. The revisions were designed to reduce the range of topics covered, to allow more depth of study and increased conceptual understanding for students. There is a shift in laboratory emphasis from instructor-designed demonstrations to student-designed investigations. While students may be introduced to concepts and methods as before, it is expected that they will develop more independent inquiry skills. Lab investigations now incorporate more student-questioning and experimental design. To accomplish this, the College Board has decreased the minimum number of required labs from 12 to 8 while keeping the same time requirement (25% of instruction time devoted to laboratory study). The College Board has defined seven science practices that students must learn to apply over the course of laboratory study. In brief, students must:

1. Use models

2. Use mathematics (quantitative skills)

3. Formulate questions

4. Plan and execute data collection strategies

5. Analyze and evaluate data

6. Explain results

7. Generalize data across domains

The College Board published 13 recommended laboratories in the spring of 2011. They can be found at: http://advancesinap.collegeboard.org/science/biology/lab

Many of these laboratories are extensions of those described in the 12 classic labs that the College Board has used in the past. The materials provided in this lab have been prepared by Ward’s to adapt to the specifications outlined in AP Biology Investigative Labs: An Inquiry-Based Approach (2012, The College Board). Ward’s has provided instructions and materials in the AP Biology Investigation series that complement the descriptions in this College Board publication. We recommend that all teachers review the College Board material as well as the instructions here to get the best understanding of what the learning goals are. Ward’s has structured each new AP investigation to have at least three parts: Structured, Guided, and Open Inquiry. Depending on a teacher’s syllabus, they may choose to do all or only parts of the investigations in scheduled lab periods.

The College Board requires that a syllabus describe how students communicate their experimental designs and results. It is up to the teacher to define how this requirement will be met. Having students keep a laboratory notebook is one straightforward way to do this.



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reCordIng dAtA In A lAborAtory notebook

All of the Ward’s Investigations assume that students will keep a laboratory notebook for student-directed investigations. A brief outline of recommended practices to set up a notebook, and one possible format, are provided below.

1. A composition book with bound pages is highly recommended. These can be found in most stationary stores. Ward’s offers several options with pre-numbered pages (for instance, item numbers 32-8040 and 15-8332.. This prevents pages from being lost or mixed up over the course of an experiment.

2. The title page should contain, at the minimum, the student’s name. Pages should be numbered in succession.

3. After the title page, two to six pages should be reserved for a table of contents to be updated as experiments are done so they are easily found.

4. All entries should be made in permanent ink. Mistakes should be crossed out with a single line and should be initialed and dated. This clearly documents the actual sequence of events and methods of calculation. When in doubt, over-explain. In research labs, clear documentation may be required to audit and repeat results or obtain a patent.

5. It is good practice to permanently adhere a laboratory safety contract to the front cover of the notebook as a constant reminder to be safe.

6. It is professional lab practice to sign and date the bottom of every page. The instructor or lab partner can also sign and date as a witness to the veracity of the recording.

7. Any photos, data print-outs, or other type of documentation should be firmly adhered in the notebook. It is professional practice to draw a line from the notebook page over the inserted material to indicate that there has been no tampering with the records.

For student-directed investigations, it is expected that the student will provide an experimental plan for the teacher to approve before beginning any experiment. The general plan format follows that of writing a grant to fund a research project.

1. Define the question or testable hypothesis.

2. Describe the background information. Include previous experiments.

3. Describe the experimental design with controls, variables, and observations.

4. Describe the possible results and how they would be interpreted.

5. List the materials and methods to be used.

6. Note potential safety issues.

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reCordIng dAtA In A lAborAtory notebook (continued)

After the plan is approved: 7. The step-by-step procedure should be documented in the lab notebook. This includes recording

the calculations of concentrations, etc., as well as the weights and volumes used.

8. The results should be recorded (including drawings, photos, data print outs, etc.).

9. The analysis of results should be recorded.

10. Draw conclusions based on how the results compared to the predictions.

11. Limitations of the conclusions should be discussed, including thoughts about improving the experimental design, statistical significance, and uncontrolled variables.

12. Further study direction should be considered.

The College Board encourages peer review of student investigations through both formal and informal presentation with feedback and discussion. Assessment questions similar to those on the AP exam might resemble the following questions, which also might arise in peer review:

• Explain the purpose of a procedural step.

• Identify the independent variables and the dependent variables in an experiment.

• What results would you expect to see in the control group? The experimental group?

• How does XXXX concept account for YYYY findings?

• Describe a method to determine XXXX.



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MAterIAls ProvIded In kIt

Units per kit

DescriptionMAterIAls needed bUt not ProvIded

Clear plastic bottles (e.g., soda bottles) with caps

1 pH Paper, 1-14 range, Vial/100Household substances, condiments,

foods with heavy odors

1 pkg. Filter Paper, Medium Grade, Clear plastic packing tape

1 Ward’s Dual Magnifier, 3X & 6X Water

1 Disposable Petri Dish, Pkg/20 Masking tape

1 Vinegar, 473 mL, White Funnel

8 Animal Behavior TraysMorgue (beaker filled with

salad oil or alcohol)

1 pkg./300 Cotton Balls Light

1 pkg./100 Pipets Alka Seltzer tablets

1 Instructions (this booklet) oPtIonAl MAterIAls ( not ProvIded)

1

Redemption coupon for pill bugs and Drosophila*

Order organisms to be delivered a week in advance of lab

Fine paintbrushes

Cold packs or crushed ice

Aluminum foil

Other materials as determined by students’ experimental design

MAterIAls CheCklIst

Call “Us” at 1.800.962.2660 for

Technical Assistance Visit “Us” on-line at

www.wardsci.com

for U.S. Customers

www.wardsci.ca

for Canadian Customers

or

* - It is recommended that you redeem your coupon for live/

perishable materials as soon as possible and specify your preferred delivery date. Generally, for timely delivery, at least a week’s advance

notice is preferred.



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CUrrICUlUM AlIgnMentBig Ideas

Big Idea 4: Biological systems interact, and these interactions possess complex properties

Big Idea 2: Biological systems utilize energy and molecular building blocks to grow, to reproduce, and to maintain homeostasis.

Enduring Understandings

2E3: Timing and coordination of behavior are regulated by various mechanisms and are important in natural selection.

2D1: All biological systems from cells and organisms to populations, communities, and ecosystems are affected by complex biotic and abiotic interactions involving the exchange of

4A6: Interactions among living systems and with their environment result in the movement of matter and energy.

4B4: Interactions between and within populations influence patterns of species distribution and abundance.

Science Practices

1.3 The student can refine representations and models of natural or man-made phenomena and systems in the domain.

2.2 The student can apply mathematical routines to quantities that describe natural phenomena.

3.2 The student can refine scientific questions.

4.2 The student can design a plan for collecting data to answer a particular scientific question.

5.1 The student can analyze data to identify patterns or relationships.

6.1 The student can justify claims with evidence.

12.2 The student can connect concepts in and across domain(s) to generalize or extrapolate in and/or across enduring understandings and/or big ideas.

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Kit # 3674-12

This lab is aligned with the 2012 AP Biology Curriculum (registered trademark of the College Board). Listed below are the aligned Content Areas (Big Ideas and Enduring Understandings), the Science Practices, and the Learning Objectives of the lab as described in AP Biology Investigative Labs: An Inquiry Approach (2012.. This is a publication of the College Board that can be found at http://advancesinap.collegeboard.org/science/biology/lab.



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leArnIng objeCtIvesThe student is able to refine scientific models and questions about the effect of complex biotic and abiotic interactions on all biological systems from cells and organisms to populations, communities, and ecosystems (2D1 & SP 1.3, SP 3.2..

The student is able to design a plan for collecting data to show that all biological systems (cells, organisms, populations, communities, and ecosystems) are affected by complex biotic and abiotic interactions (2D1 & SP 4.2, SP 7.2..

The student is able to analyze data to identify possible patterns and relationships between a biotic or an abiotic factor and a biological system (cells, organisms, populations, communities, or ecosystems) (2D1 & SP 5.1..

The student is able to analyze data to support the claim that response to information and communication of information affect natural selection (2E3 & SP 5.1..

The student is able to justify claims, using evidence, to describe how timing and coordination of behavioral events in organisms are regulated by several mechanisms (2E3 & SP 6.1..

The student is able to connect concepts in and across domain(s) to predict how environmental factors affect response to information and change behavior (2E3 & SP 7.2..

The student is able to apply mathematical routines to quantities that describe interactions among living systems and their environment that result in the movement of matter and energy (4A6 & SP 2.2..

The student is able to use visual representations to analyze situations or solve problems qualitatively to illustrate how interactions among living systems and with their environments result in the movement of matter and energy (4A6 & SP 1.4. .

The student is able to predict the effects of a change of matter or energy availability on communities (4A6 & SP 6.4. .

The student is able to use data analysis to refine observations and measurements regarding the effect of population interactions on patterns of species distribution and development (4B4 & SP 5.2..

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tIMe reqUIreMents

Pre-Lab Prep: Redeem Live Materials Coupon

At least 1 week prior to lab

Part 1: Structured Inquiry – Chemotaxis

Total of 30 minutes*: 10 minutes set up 10 minutes observation and recording 10 minutes analysis

*Optional: Teacher may decide to break this into two observation periods – 30 min-utes for control (no stimulus in tray) and 30 minutes for experimental)

Part 2: Guided Inquiry – Test Variables of Student’s Choice

Total of 20 minutes: 5 minutes set up 10 minutes observation and recording 5 minutes analysis

Part 3: Open InquiryTotal depends on student/teacher scheduling and parameters of experiment



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CheMICAl sAfety Review all Material Safety Data Sheets (MSDSs) for all precautions, handling procedures, storage, and information.

Review local regulations or consult with local authorities before disposing of any chemicals in the trash or down the drain. Instruct your students on the proper disposal procedure for all leftover reagents and products from the laboratory activity.

When working in the laboratory, especially with chemicals, we recommend the use of personal protective equipment, including safety goggles (ANSI-approved), chemical resistant laboratory apron, and gloves.

In the event of a chemical spill or mishap, follow the procedures outlined in the appropriate MSDS for cleanup or corrective action. Contact any local authorities when necessary.

generAl sAfety PreCAUtIons The teacher should be familiar with safety practices and regulations in their school (district and state). Know what needs to be treated as hazardous waste and how to properly dispose of non-hazardous chemicals or biological material.

Consider establishing a safety contract that students and their parents must read and sign off on. This is a good way to identify students with allergies to things like latex so that you (and they) will be reminded of what particular things may be risks to individuals. A good practice is to include a copy of this contract in the student lab book (glued to the inside cover).

Students should know where all emergency equipment (safety shower, eyewash station, fire extinguisher, fire blanket, first aid kit etc.) is located.

Make sure students remove all dangling jewelry and tie back long hair before they begin.

Remind students to read all instructions, MSDSs, and live care sheets before starting the lab activities and to ask questions about safety and safe laboratory procedures. In most of these AP Biology labs, appropriate MSDSs and live care sheets can be found on the last pages of this booklet. Additionally, the most updated versions of these resources can be found at: www.wardsci.com , under Living Materials http://wardsci.com/article.asp?ai=1346.

In student directed investigations, make sure that collecting safety information (like MSDSs) is part of the experimental proposal.

At end of lab:

All laboratory bench tops should be wiped down with a 20% bleach solution or disinfectant to ensure cleanliness.

Remind students to wash their hands thoroughly with soap and water before leaving the laboratory.

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Pre-lAborAtory PrePArAtIonRedeem your coupon for all live materials to be delivered at least one week before you are ready to start this lab.

This kit contains redemption coupons for both Drosophila and pillbugs. Either organism can be used throughout the lab. It is recommended that you provide both for your students so they have expanded choices for constructing their own experiments. If you would like to assist students in making a “choice chamber” with two sections, instead of the behavior tray with five sections provided here, please see the instructions below. Students may find the two-chambered setup easier for performing geotactic experiments.

Have students read the experiment ahead of time and have them bring in an approved stimulus of their choice for the Guided inquiry part of the lab.

If using Drosophila : Place vial in refrigerator or in ice bucket at beginning of class so that the animals will be moving slowly enough to handle by the time students need to fill the behavior tray. Leaving the Drosophila in the cold for more than 30 minutes will decrease their viability.

Prepare an insect morgue to kill and dispose of insects: Typically, this is a beaker or flask containing either salad oil or 70% alcohol. Once the insects are dead, dispose of morgue contents as recommended by your school. Generally, contents can be flushed down a sink drain (if using oil- treat with dishwashing liquid/soap prior to pouring down the drain) with copious amounts of water.

OPTIONAL : Prepare solutions of household materials for students to use in guided inquiry in stock bottles. Only allow students access to dropper bottles or other small bottles of solutions. Clearly label all bottles. Lab solutions of HCl or NaCl should be no more concentrated than 0.1 M. Volatile choices might include alcohol (associated with fermentation), ammonia (nitrogen associated with decay), mercaptoethanol (sulphur associated with decay), soil with high humus content, wet yeast (associated with fermentation), apple cider vinegar, fruit juice, cedar chips.

OPTIONAL - Construct a 2 choice chamber as an alternative to the 4 choice chamber provided:

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PreP tIP

In this lab you will use live materials. Prior to starting this lab, submit your live/perishable material redemption coupon via mail, fax, or simply calling into customer service at 1-800-962-2660. It is recommended that you do this at least one week before the lab.

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Have students bring in clear, soft-plastic water bottles,12-16 oz in size. For eight lab groups, collect at least 16 bottles with caps. Pair the water bottles by size (not all bottles must be the same size, but each 2-chamber unit must have the same-sized chambers). Have students cut off the bottom of the bottles using scissors clean and dry the bottles thoroughly. Match the bottles end to end and tape them together using clear packaging tape. Label one side “A” and the other side “B.” When testing substances, cotton balls are soaked in the testing material, inserted into either chamber, and the caps are screwed on tightly. Be sure to clean the bottles, bottle caps and bottle necks between tests.

before ClAss

1. Make copies of the Student Guide (pages 1 - x) and additional laboratory sheets.

2. Prepare your materials for the class demonstration to support your lecture or laboratory introduction.

Pre-lAborAtory PrePArAtIon (ContInUed)oPtIonAl Pre-lAb deMonstrAtIon

Given the definition of “taxis,” what do students think “geotaxis” refers to? Demonstrate geotaxis by having students observe a vial of Drosophila. Note what direction the flies move in. After a minute, invert the vial. Did the flies’ behavior change? Why do students think this occurs?

Give an example of data and results, including analysis of error (overlapping error – not statistically significant, non-overlapping-significant). Discuss T-test and chi squared test.

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bACkgroUndOrganisms orient to stimuli that are important to their survival. Movement toward or away from important stimuli depends upon both the sensory and motor abilities of the organism. A stimulus might involve anything that can be sensed, like light, sound, touch, heat, or chemicals. For example, humans do not sense a magnetic field and cannot orient towards it (without instruments like a compass). Therefore, we may infer that magnetic fields have not been very important for human survival as a species. Once an organism with sensory/motor abilities perceives a stimulus, it can orient or move either towards or away from that stimulus depending upon the nature of the stimulus (opportunity or threat). Movement in response to a stimulus is classified as taxis, whereas random movement or movement irrespective of stimulus is classified as kinesis. Generally, the more critical a stimulus is to an organism’s survival, the stronger the response to that stimulus. Therefore, an organism that senses an optimal food source will usually orient strongly toward it. In an animal that senses primarily through smell, movement towards an appropriate food sourcewould be called positive chemotaxis. In the same vein, orienting with reference to light is called phototaxis, and orienting in response to gravity is called geotaxis, etc.

Behavior can be classified as innate or learned. Innate behavior is inherited and instinctive, and develops independently of the experience of an organism in its environment over time. On the other hand, learned behaviors are not inherited and can be changed as a result of the animal’s experience with its environment and other organisms.

In this laboratory, you will be investigating and observing the taxis and kinesis of model organisms- either fruit flies or pill bugs. As you make behavioral observations, think about how those behaviors contribute to making the species an evolutionary success in its natural environment. Since differential reproduction is a strong driver of evolution, you may want to take special note of any taxis related to reproductive behaviors.

Drosophila melanogaster represents a model organism with well analyzed genetics and many mutant strains available (see www.fruitfly.org or www.flybase.org as well as the care sheet in this booklet). Further, this organism has several distinct developmental stages that can be investigated separately (larval, pupae, and adult). Larval stages are advantageous to study since the organisms are slow moving and do not fly, however, they may be too slow to make relevant behavioral investigations in the supplied behavioral tray (students may want to

objeCtIves

Refine scientific models and questions about the effect of complex biotic and abiotic interactions on all biological systems from cells and organisms to populations, communities, and ecosystems.

Design a plan for collecting data to show that all biological systems (cells, organisms, populations, communities, and ecosystems) are affected by complex biotic and abiotic interactions.

Analyze data to identify possible patterns and relationships between a biotic or an abiotic factor and a biological system (cells, organisms, populations, communities, or ecosystems).

Analyze data to support the claim that response to information and communication of information affect natural selection.

Justify claims, using evidence, to describe how timing and coordination of behavioral events in organisms are regulated by several mechanisms.

Connect concepts in and across domain(s) to predict how environmental factors affect response to information and change behavior.

Apply mathematical routines to quantities that describe interactions among living systems and their environment that result in the movement of matter and energy.

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(continued on next page)(continued on next page)



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consider making modifications). Pupae do not display behavior, and adults are very fast moving, and since they fly it is challenging to get the specimens into and out of the supplied behavioral tray. Cooling the vial of flies for 10-20 minutes in a refrigerator or on ice or a cool pack will slow the adults down enough to be handled more easily. Fine paintbrushes are an excellent tool for handling adults that have been slowed. Adult male Drosophila are distinguishable from females in that they are smaller than females, and they have dark sexcombs on their first (most anterior) pair of legs. Drosophila will eat many different fruits and vegetables.

Isopods (pill bugs or sow bugs) do not have a bank of mutants available for behavioral experiments but they do provide the advantage of being easy to handle in the context of these behavioral experiments. They also represent organisms with a different evolutionary history, and they are adapted to environments that overlap with fruit fly environments. Isopods, however, are distinct and have evolved different developmental patterns as well as different sensory and motor responses to stimuli. To determine the sex of an isopod use a stereomicroscope or magnifying glass to observe the underside of the specimen near the posterior end. Males have two white, elongated appendages that serve as copulatory organs. These are modified “pleopods” and are absent in the females. Pill bugs and sowbugs eat decaying plant material.

objeCtIves (ContInUed)

Use visual representations to analyze situations or solve problems qualitatively to illustrate how interactions among living systems and with their environments result in the movement of matter and energy.

Predict the effects of a change of matter or energy availability on communities.

Use data analysis to refine observations and measurements regarding the effect of population interactions on patterns of species distribution and development.

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bACkgroUnd (ContInUed)



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sAfety PreCAUtIons As general safe laboratory practice, it is recommended that you wear proper protective equipment, such as gloves, safety goggles, and a lab apron.

As general lab practice, read the lab through completely before starting, including any Material Data Safety Sheets (MSDSs) and live materials care sheets at the end of this booklet as well as any appropriate MSDSs for any additional substances you would like to test. One of the best sources is the vendor for the material. For example, when purchased at Ward’s, searching for the chemical on the Ward’s website will direct you to a link for the MSDS.

At the end of the lab:

All laboratory bench tops should be wiped down with a 20% bleach solution or disinfectant to ensure cleanliness.

Wash your hands thoroughly with soap and water before leaving the laboratory.

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notes



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ProCedUre tIPs

When performing this lab, all data should be recorded in a lab notebook. You will need to construct your own data tables, where appropriate, in order to accurately capture the data from the investigation.

Record all data ImmEDIAtEly in your laboratory notebook.

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PArt 1 – CheMotAxIs (strUCtUred InqUIry)

ProCedUre –CheMotAxIsYour teacher will assign you an organism, either fruit flies or pill bugs, to observe and investigate its behaviors.

1. To become familiar with the organisms, sketch an illustration of your organism in your laboratory notebook. Label all anatomical structures that you recognize. Can you differentiate male from female?

2. Place one piece of masking tape on the outside of each chamber of the behavior tray; label them A, B, C, D, and E (central chamber).

3. Place a drop or two of vinegar on a cotton ball or on a small section of filter paper and place this in chamber A (not central chamber). Plain water on a similar paper in the opposite chamber can provide one control condition.

4. Place several (2 or fewer in each chamber for a total of 10. organisms in the behavior tray, cover the tray with clear cover, and carefully observe the specimens for at least 10 minutes. (NOTE: If you are using fruit flies, place them in the refrigerator for five minutes before they are needed. They will be easier to deposit into the behavior trays as the cooler temperature slows them down. Use funnel if necessary.) Document any behaviors you see in a list. Remember to record even seemingly unimportant behaviors.

5. Every minute for 10 minutes, count the number of organisms in each chamber and record observations in Table 1 (on next page).

Do not disturb the pill bugs or fruit flies; shaking or tipping the tray will introduce additional stimuli.

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6. Graph your results in the space below.

ProCedUre –PArt 1: CheMotAxIs (ContInUed)


table 1: Organism taxis

Time (min)

# Organisms in Chamber A vinegar

# Organisms in Chamber

B


CWater

(opposite A)


D


E central

AverageClass

Average

0 2 2 2 2 212345678910

Ave.Class avg.



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notes 7. Calculate the average number of organism in each chamber in the 10-minute period of time. Add this to Table 1.

8. Using the data from every group in the class, calculate the class average for number of organisms in each chamber in a 10-minute time period. Enter this data in Table 1. Calculate standard error and deviation. Draw a graph to discuss whether the results are significantly different between empty chambers and vinegar chambers. You may want to use data points only after 5 minutes for comparisons to allow for taxis to occur. If two types of organisms were used in class – compare whether there were significant differences between organisms.

At end of lab:

• Fruit flies and pill bugs are living organisms that should not be released to the environment. After all the investigations are complete, organisms should be tapped into a “morgue” through a funnel. The morgue typically is a 150-mL beaker that contains about 50 mL of salad oil or 70% alcohol.

• All laboratory bench tops should be wiped down with a 20% bleach solution or disinfectant to ensure cleanliness.

• Wash your hands thoroughly with soap and water before leaving the laboratory.

ProCedUre –PArt 1: CheMotAxIs (ContInUed)



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1. Graph vinegar chamber over time vs control chamber over time.

2. Graph bar graphs with error bars for each chamber.

3. Was there positive or negative chemotaxis?

Adult and larval Drosophila would be expected to have strong positive chemotaxis to vinegar since it is associated with decaying fruit- the natural food source. Isopods may have a somewhat weaker positive chemotactic response since their more common food is decaying wood that would not produce strong vinegar smell.

4. What uncontrolled sensory factors may have affected (artificially skewed) results? How might you control for those factors?

The direction of light or shadows of students making observations may have affected behavior in addition to the chemosensory stimulus. The experiment can be repeated with tray in different orientation or pooling classroom results where experiments were done in many orientations should statistically control for this variable.

5. Why would sensing and responding to vinegar be important to an organism?

Ifthesmellisstronglyassociatedwithapreferredfoodsource,thisabilitywouldinfluencesurvival of the adult, as well as offspring that would hatch in the middle of a good food source.Pillbugsmaybedifferentlyresponsivethanfruitfliesduetodifferentfoodpreferences.

6. How would natural selection affect an organism’s ability to sense and/respond to this stimulus?

Differential reproduction would strongly select for individuals that had the best ability to detectagreatfoodsourceandlayeggsinthatfoodsource.Individualsthatcouldnotfindthefood sources as well would not leave as many viable progeny and would therefore be selected against.

What other types of stimuli would be important for the organism to sense and respond to?

AssessMent - PArt 1: CheMotAxIs




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7. What other types of stimuli would be important for the organism to sense and respond to?

Stimuli associated with food, sex, predators, toxicity

Chemoattractants associated with mating - locating the opposite sex

Chemorepellents associated with toxicity or predators

Light levels – might indicate where the food is or the predators are not

Sounds associated with mating- healthy males often produce the sounds that are most attractive to females

Detection of air currents associated with a predator strike

AssessMent - PArt 1: CheMotAxIs (ContInUed)



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PArt 2 – gUIded InqUIry: envIronMentAl fACtor of stUdent’s ChoICe

ProCedUre1. Identify a stimulus you would like to test for chemotaxis on your

organism. Household items or foods that have a noticeable odor, and/or volatile chemicals associated with decay or fermentation might be good choices.

2. Put this stimulus in the chamber opposite the vinegar stimulus.

3. Introduce all 10 organisms into the central chamber and cover.

4. Observe every minute for 10 minutes and record observations and analyze as described in Part 1.

ProCedUre tIPs

When performing this lab, all data should be recorded in a lab notebook. You will need to construct your own data tables, where appropriate, in order to accurately capture the data from the investigation.

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PArt 2 – AssessMent: gUIded InqUIry1. What conclusions can you make about the new stimulus relative to the vinegar stimulus?

Stronger or weaker is more or less important – confounding factors would be relative concentration.

2. How might this relate to relative selective pressures on the organism?

Stronger taxis is more closely associated with a stronger pressure.

3 Are the responses documented here innate behaviors or learned behaviors? What evidence do you have for this? What evidence would definitely indicate this?

Innate, parents aren’t usually around young to teach, Maturation of young in isolation preservesbehaviororIfyoucouldfindamutant(notdirectlyassociatedwithsenseormotorfunction)thatdisruptedorintensifiedthebehavior.

4. Create a table for all the results you have obtained. Indicate whether the household substances tested are equally attractive or repellant to the pill bugs or fruit flies. Identify patterns in their behavior.

Answers will vary depending on the individual student investigations.



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PArt 3: oPen InqUIry: desIgn An exPerIMentWhat questions occurred to you as you observed the taxic behaviors of your organism? Now that you are familiar with the organism and how to test for taxis, design an experiment to investigate one of your questions. You may want to expand on your experiment from Part B by creating different concentrations of your material, or you may wish to begin a new investigation. Other factors you can test include pH, temperature, light intensity or wavelength, substrate composition. You may want to look at sensing and taxis as it is related to reproductive success or you may want to specifically compare organisms from overlapping environmental niches to test hypotheses about the relative differences in selective pressures.

Before starting your experiment, have your teacher check over, approve, and initial your experiment design. Once your design is approved, investigate your hypothesis. Be sure to record all observations and data in your laboratory sheet or notebook.

Use the following steps when designing your experiment.

1. Define the question or testable hypothesis.

2. Describe the background information. Include previous experiments.

3. Describe the experimental design with controls, variables, and observations.

4. Describe the possible results and how they would be interpreted.

5. List the materials and methods to be used.

6. Note potential safety issues.

After the plan is approved by your teacher:

7. The step by step procedure should be documented in the lab notebook. This includes recording the calculations of concentrations, etc. as well as the weights and volumes used.

8. The results should be recorded (including drawings, photos, data print outs).

9. The analysis of results should be recorded.

10. Draw conclusions based on how the results compared to the predictions.

11. Limitations of the conclusions should be discussed, including thoughts about improving the experimental design, statistical significance and uncontrolled variables.

12. Further study direction should be considered.

exPerIMent desIgn tIPs

The College Board encourages peer review of student investigations through both formal and informal presentation with feedback and discussion. Assessment questions similar to those on the AP exam might resemble the following ques-tions, which also might arise in peer review:

Explain the purpose of a procedural step.

Identify the independent variables and the dependent variables in an experiment.

What results would you expect to see in the control group? The experimental group?

How does XXXX concept account for YYYY findings?

Describe a method to determine XXXX.

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lIve MAterIAl CAre sheet

US: P.O. Box 92912 • Rochester, NY • 14692-9012 | 812A Fiero Lane • San Luis Obispo, CA 93401 • 800-962-2660Canada: 399 Vansickle Road • St. Catharines, ON L2S 3T4 • 800-387-7822www.wardsci.com

Conditions for Customer Ownership We hold permits allowing us to transport these organisms. To access permit conditions, click here.Never purchase living specimens without having a disposition strategy in place. There are currently no USDA permits required for this organism. However, Drosophila are a pest. While it is permitted to keep them forstudy and to raise them as a food source for other animals, they should never be released into the wild.

Primary Hazard ConsiderationsAlways wash your hands thoroughly before and after you handle your Drosophila, their food, or anything they have touched.

AvailabilityStandard strains of Drosophila are available year round as they are lab raised. Drosophila come in vials with a white piece of netting andDrosophila media for food, and a foam cap. We over-pack each order of Drosophila. It is normal to have some deceased Drosophila in thecontainer. You will receive at least the quantity of live Drosophila stated on the container. There should be between 30 and 50Drosophila in the container. Drosophila can survive in this container for about three weeks. Drosophila should be kept at room temper-ature (65–75°F).

Drosophila crosses should be ordered 10 days in advance to insure that enough virgin flies are available to make the crosses. Thereshould be 5–10 original parent flies in the vial. The F1 generation (the result of the cross) is in the media as eggs and larvae. When the larvae are visible, remove both surviving and dead parents to avoid counting them as part of the F1 generation.

Standard Drosophila crosses contain the following flies:

Species: melanogasterGenus: Drosophila Family: Drosophilidae Order: DipteraClass: InsectaPhylum: ArthropodaKingdom: Animalia

DrosophilaFruit Fly

Cross Female x MaleMonohybrid A Sepia x Wild

Dihybrid B Vestigial x SepiaSex-Linked C White x Wild




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Name Description/Characteristics/Notes

Drosophila melanogaster, Normal characteristics (red eyes and normal wing size).Wild type (+)

Drosophila mojavensis Long life cycle (approximately one month).Useful for comparative population studies and for preparing salivary gland chromosome squashes.

Drosophila virilis One of the largest Drosophila species. Useful for preparing salivary gland chromosome squashes.

Chromosome I Mutants (Sex-Linked)

Apricot (a) Apricot eyes, males’ eyes slightly larger than females’.

Bar (B) Bar eyes (narrow vertical bar), males’ eyes larger than females’.

Forked (f) Forked bristles.

Miniature (m) Miniature wings (slightly longer than abdomen, but with normal proportions); dark gray and less transparent than normal.

Ruby (b) Clear ruby eyes.

Vermilion (v) Vermilion (bright red) eyes.

White (w) White eyes.

White-Crossveinless-Forked White eyes, crossveinless wings, forked bristles.(w; cv; f)

White-Miniature-Forked White eyes, miniature wings, forked bristles.(w; m; f)

Yellow (y) Lightest body color.

Yellow-Forked-Attached, Females: normal eyes, yellow body with forked bristles, normal wings.and White (yf:=&w[1]) Males: white eyes, gray body, normal wings.

Note: Remove culture from stock when all females have died or when all flies have white eyes.

Yellow-White (y;w) Lightest body color, white eyes.

Yellow-White-Miniature Lightest body color, white eyes, miniature wings.(y;w; m)

Chromosome II Mutants

Apterous (ap) Wingless (lacking all wing blade structures).

Black (b) Black body (darker at cooler temperatures).

Black-Purple-Curved (b;pr;c) Black body, purple eyes (ruby upon emergence, darken to purple with age), curved wings.

Black-Vestigial (b;vg) Black body, vestigial wings.

Brown (bw) Brown eyes (light brown/wine upon emergence, darken to garnet with age). Produces white eyeswhen crossed with vermilion, cinnabar, or scarlet.

Cinnabar (cn) Cinnabar eyes (slightly brighter than wild, dull with age).

Dumpy (dp) Truncated wings (approximately half normal length).



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Chromosome II Mutants (continued)

Heldout (ho) Wings extended at right angles to body.

Lobe (L) Lobe-shaped eyes (slightly reduced size with a nick in anterior edge;lower half reduced more than upper half).

Star-Curly (S;Cy) Eyes rough, slight smaller and narrower than wild type. Wings curved upward.

Vestigial (vg) Vestigial wings (increase in size at 29°C or higher).

Vestigial-Brown (vg;bw) Vestigial wings, brown eyes (darken with age).

Chromosome III Mutants

Antennapedia (Antp) Legs grow in place of antennae.

Ebony (e) Shiny black body.

Scarlet (st) Scarlet (eyes bright vermilion; darken with age).

Sepia (se) Sepia eyes (brown at emergence, darken to sepia, become black with age).

Sepia-Ebony (se;e) Sepia eyes, shiny black body.

Chromosome IV Mutants

Eyeless (ey) Eyes half to one-quarter normal size.

Polished (pol) Eyes without facets (glasslike).

Multichromosomal Mutants

Curly/Plum (II); Curly wings, plum eyes, wings divergent, bristles short and thick. Dichaete/Stubble (III) Easily prone to contamination; subculture frequently.(Cy/Pm;D/sb)

Dumpy (II); Sepia (III) Truncated wings, sepia eyes.(dp;se)

Vestigial (II); Sepia (III) Vestigial wings, sepia eyes.(vg;se)

Vestigial (II); Ebony (III) Vestigial wings, shiny black body.(vg;e)

Yellow-Forked-Attached, Females: normal eyes, yellow body, apterous wings. Males: white eyes, gray body, apterous wings.and White (I); Apterous (II) Note: Remove culture from stock when all females have died or when all flies have white eyes.(yf:=&w[1];ap[2])



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© 2008 Ward’s Natural Science Establishment. All rights reserved. Rev. 4/10

Captive CareHabitat:Any escape-proof container with holes for oxygen exchange (we use plastic vials with foam tops). Drosophila should be kept at roomtemperature (65–75°F). Care:

• Food: Drosophila will eat many different fruits and vegetables. The simplest is just a slice of banana. Many different types of culture media have been developed. Each formulation has benefits and shortcomings. In order for a medium to be useful, it mustbe solid and dry enough so the adult flies do not stick to or drown in, yet moist enough to provide water and it must inhibit thegrowth of ubiquitous environmental mold. Subculture when Media 38 W 0592 is depleted, drying out, or has developed mold.Provide about an inch of dry media at the bottom of the Culture Vial 18 W 4956, and about 15 mL of water.

Information• Method of Reproduction: Sexual. Male inseminates female for internal fertilization and the female then lays eggs, about 100

per day. The sperm received by a female fly during mating is retained, serving to fertilize a number of eggs. Therefore, in anexperimental cross between two different strains, virgin females must be used to insure that the expected genetic cross producesthe resultant eggs. NOTE: Since females do not become sexually mature for about 10 hours, virgin females can be collected within10 hours of removing all adult flies from a culture with pupae. Virgin flies may lay eggs, but they will not hatch.

Life CycleDevelopment time is influenced by both temperature and strain. At room temperature, it takes about 24 hours for an egg to hatch intoa larva. The larva feeds as it burrows through the medium. As the larva grows, it undergoes two molts so that the larval period con-sists of three stages (instars), the first instar being the newly hatched larva. The first two instars last about 24 hours each. The finallarval stage or third instar lasts about 48 hrs and the larva may attain a length of 4.5 mm. Towards the end of the third instar stage,the larva will crawl up the sides of the culture jar, attach itself to a dry surface (the jar, filter paper, etc.) and form the pupa. Afterabout four days in the pupal stage, an adult fly emerges. Females become sexually mature 8–10 hours after emerging from the pupae.Adults will live for about 30 days.

Wild HabitatMany different species of Drosophila are present throughout the world. Drosophila melanogaster is primarily used for genetic studiesfor a variety of reasons. Its entire genome has now been sequenced. In the wild, Drosophila melanogaster is thought to have originatedin tropical regions of the eastern hemisphere. They can now be found throughout the world with the exception of the arctic.Drosophila are pests that infest rotted fruits and fruits that are beginning to rot. They are also attracted to vinegar and wine. Drosophilacan destroy harvested crops in storage. They are food for hummingbirds, frogs and reptiles.

Disposition• We do not recommend releasing any laboratory animal into the wild, and especially not insects that are considered to be pests or

not native to the environment. • Adoption is the preferred disposition for any living animal. • If the insects must be euthanized at the end of study, follow one of these procedures:

• Put them into a container or bag and freeze for 48 hours. • Place the organism in 70% isopropyl alcohol for 24 hours.• Autoclave the organism @ 121°C for 15 minutes.

• A deceased specimen should be disposed of as soon as possible. Consult your school’s recommended procedures for disposal. In general, dead insects should be handled as little as possible or with gloves, wrapped in an opaque plastic bag that is sealed(tied tightly) before being placed in a general garbage container away from students.



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Conditions for Customer Ownership We hold permits allowing us to transport these organisms. To access permit conditions, click here.Never purchase living specimens without having a disposition strategy in place. There are currently no USDA permits required for this organism. In order to protect our environment, never release a live laboratoryorganism into the wild.

Primary Hazard ConsiderationsAlways wash your hands thoroughly after you handle your organism.

AvailabilityLand isopods supplied are either Sowbugs 87 W 5520 (Oniscus or Porcellio), package of 45 or Pillbugs 87 W 5525 (Armadillium), package of 45 and are generally available year-round, but they are wild-collected so shortages may occur. Substitutions of one speciesfor the other may occur during shortages. Land isopods are shipped in plastic food containers with damp sphagnum moss or papertowel and a piece of potato or carrot. We over-pack each order of isopods. It is normal to have some deceased isopods in the container.You will receive at least the quantity of live isopods stated on the container. Average size of the land isopod is 1.0 centimeters. Theisopods can survive up to one week in the shipping container.

Captive CareHabitat:

• Use a small aquarium or plastic storage box to house your land isopods. The container should have a perforated lid to permit airexchange, but perforations should be small enough to prevent escape. Cover the floor of the container with five centimeters ofrich soil. The soil in the habitat should be kept moist but not wet. Check daily and mist if necessary. Add stones, pieces of bark, or crumpled paper to the habitat to provide cover for the isopods.

• Maintain the habitat at room temperature. Soil should be replaced once or twice a year and any dead isopods should be removedfrom the container.

Care:• Add a slice or two of potato per 50 pillbugs, which will provide the isopods with both food and moisture. Replace the

potato weekly.

Information• Method of reproduction: Sexual• Determining Sex: On the underside of the female isopod, leaf-like growths can be observed at the base of some of the legs. These

are known as brood pouches. On male isopods, the first two appendages on the abdomen are modified as elongated copulatoryorgans.

• Pillbugs have the ability to curl up into a ball when they feel threatened. Sowbugs cannot roll up into a ball.

Genus: Oniscus, Porcellio or ArmadilliumFamily: OniscoideaOrder: IsopodaClass: MalacostracaSubphylum: CrustaceaPhylum: ArthropodaKingdom: Animalia

Land IsopodsPillbugs and Sowbugs


Determining Sex: Using a stereomicroscope or magnifying glass, observe the underside of the isopod near the posterior end. Males have two white, elongated appendages that serve as copulatory organs. These are modified “pleopods” and are absent in the females.



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© 2008 Ward’s Natural Science Establishment. All rights reserved. Rev. 9/08, 11/09

Life Cycle• Unlike other crustaceans which live on land, land isopods do not need to lay their eggs in an aquatic environment. Instead, the

eggs of land isopods are brooded in a fluid-filled pouch on the underside of the female. After approximately three weeks, up to200 young isopods, which are similar in appearance to the adults, emerge from the marsupium. They molt several times to grow.The entire life cycle takes two to three months.

Wild HabitatLand isopods generally inhabit damp, dark environments such as gardens and woodlands where they hide under stones and logs. Landisopods are nocturnal, coming out at night to feed on plant matter and decayed wood.

Disposition• We do not recommend releasing any laboratory animal into the wild, and especially not invertebrates that are not native to the

environment.• Adoption is the preferred disposition for any living animal. • If the isopods must be euthanized at the end of study, follow one of these procedures:

• Put them into a container or bag and freeze for 48 hours. • Place the organism in 70% isopropyl alcohol for 24 hours.

• A deceased specimen should be disposed of as soon as possible. Consult your school’s recommended procedures for disposal. In general, dead insects should be handled as little as possible or with gloves, wrapped in an opaque plastic bag that is sealed(tied tightly) before being placed in a general garbage container away from students.




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MAterIAl sAfety dAtA sheetMaterial Safety Data Sheet Page 1 of 2

© 2009, Scholar Chemistry. All Rights Reserved. 1/20/2012

Vinegar MSDS # 786.50

Section 1: Product and Company Identification Vinegar

Synonyms/General Names: Acetic acid, Ethanoic acid. Product Use: For educational use only. Not for human consumption.Manufacturer: Various

24 Hour Emergency Information Telephone Numbers CHEMTREC (USA): 800-424-9300 CANUTEC (Canada): 613-424-6666

ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com

Section 2: Hazards Identification Clear, colorless solution with a strong vinegar odor. HMIS (0 to 4)

CAUTION! Body tissue irritant and slightly toxic by ingestion. Not for human consumption Target organs: Respiratory system, eyes, skin, teeth.

This material is considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200).

Section 3: Composition / Information on Ingredients Acetic Acid (64-19-7), 4-6%. Water (7732-18-5), 94-6%.

Section 4: First Aid Measures Always seek professional medical attention after first aid measures are provided.

Eyes: Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.Skin: Immediately flush skin with excess water for 15 minutes while removing contaminated clothing. Ingestion: Call Poison Control immediately. Do not induce vomiting. Rinse mouth with cold water. Give victim 1-2 cups of

water or milk to drink. Inhalation: Remove to fresh air. If not breathing, give artificial respiration.

Section 5: Fire Fighting Measures When heated to decomposition, emits acrid fumes of carbon oxides. 0 Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire. 0 0

Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool container with water spray. Material is not sensitive to mechanical impact or static discharge.

Section 6: Accidental Release Measures Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected personnel. Remove all ignition sources and ventilate area. Contain spill with sand or absorbent material and place material in a sealed bag or container for disposal. Wash spill area after pickup is complete. See Section 13 for disposal information.

Section 7: Handling and Storage White Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash

hands thoroughly after handling. Storage: Store in Corrosive Area [White Storage] with other corrosive items. Store in a dedicated corrosive cabinet. Store in a

cool, dry, well-ventilated, locked store room away from incompatible materials.

Section 8: Exposure Controls / Personal ProtectionUse ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash hands thoroughly after handling material and before eating or drinking. Use NIOSH-approved respirator with an acid/organic cartridge. Exposure guidelines: Acetic Acid: OSHA PEL: 25 mg/m3 and ACGIH: 10 ppm TLV, 15 ppm as STEL.

Health 1 Fire Hazard 0 Reactivity 0




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MAterIAl sAfety dAtA sheet

Material Safety Data Sheet Page 2 of 2

© 2009, Scholar Chemistry. All Rights Reserved. 1/20/2012

MSDS # 786.50 Vinegar Scholar Chemistry

Section 9: Physical and Chemical Properties Molecular formula CH3COOH. Appearance Clear, colorless liquid.Molecular weight 60.05. Odor vinegar.Specific Gravity 1.00 g/mL @ 20°C. Odor Threshold 0.48 ppm. Vapor Density (air=1) N/A. Solubility Completely soluble in water. Melting Point N/A. Evaporation rate N/A (Butyl acetate = 1). Boiling Point/Range N/A. Partition Coefficient N/A (log POW). Vapor Pressure (20°C) N/A. pH 5, acidic. Flash Point: N/A. UEL N/A. Autoignition Temp.: N/A. LEL N/A.

N/A = Not available or applicable

Section 10: Stability and Reactivity Stability: Stable under normal conditions of use and storage. Avoid heat and ignition sources. Incompatibility: Oxidizing agents, metals, soluble carbonates and phosphates, hydroxides, amines, and alcohols Shelf life: Indefinite if stored properly.

Section 11: Toxicology Information Acute Symptoms/Signs of exposure: Eyes: Redness, tearing, itching, burning, damage to cornea, conjunctivitis, loss of vision. Skin: Redness, blistering, burning, itching, tissue destruction with slow healing. Ingestion: Nausea, vomiting, burning, diarrhea, ulceration, convulsions, shock. Inhalation: Coughing, wheezing, shortness of breath, headache, spasm, inflammation and edema of bronchi, pneumonitis. Chronic Effects: Repeated/prolonged skin contact may cause thickening, blackening or cracking. Repeated eye exposure may cause corneal erosion or loss of vision. Sensitization: none expected Acetic acid: LD50 [oral, rat]; 3310 mg/kg; LC50 [rat]; >16000 (4 hour); LD50 Dermal [rabbit]; 1120 mg/kg Material has not been found to be a carcinogen nor produce genetic, reproductive, or developmental effects.

Section 12: Ecological Information Ecotoxicity (aquatic and terrestrial): Not available

Section 13: Disposal Considerations Check with all applicable local, regional, and national laws and regulations. Local regulations may be more stringent than regional or national regulations. Small amounts of this material may be suitable for sanitary sewer disposal after being neutralized to pH 7.

Section 14: Transport Information DOT Shipping Name: Not regulated by DOT. Canada TDG: Not regulated by TDG. DOT Hazard Class: Hazard Class: Identification Number: UN Number:

Section 15: Regulatory Information EINECS: Listed (200-580-7). WHMIS Canada: Not WHMIS controlled. TSCA: All components are listed or are exempt. California Proposition 65: Not listed.The product has been classified in accordance with the hazard criteria of the Controlled Products Regulations and the MSDS contains all the information required by the Controlled Products Regulations.

Section 16: Other Information Current Issue Date: January 20, 2012 Disclaimer: Scholar Chemistry and Columbus Chemical Industries, Inc., (“S&C”) believes that the information herein is factual but is not intended to be all inclusive. The information relates only to the specific material designated and does not relate to its use in combination with other materials or its use as to any particular process. Because safety standards and regulations are subject to change and because S&C has no continuing control over the material, those handling, storing or using the material should satisfy themselves that they have current information regarding the particular way the material is handled, stored or used and that the same is done in accordance with federal, state and local law. S&C makes no warranty, expressed or implied, including (without limitation) warranties with respect to the completeness or continuing accuracy of the information contained herein or with respect to fitness for any particular use.

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