Biology Lab report
Jimmonika
lab_report_instruction.docxWHAT’S IN YOUR WATER?
We take for granted the water we use to brush our teeth, wash our clothes and keep our lawns green. However, water is an extremely limited resource on the planet and we need to conserve and protect it from pollution. Today you will learn how the levels of chemicals (natural and added) are measured in different sources of water and learn whether they are a danger to your health.
LEARNING OBJECTIVES
In this laboratory you will learn:
· To identify what factors can contribute to water quality
· To use negative and positive controls to validate experimental results
· To use laboratory techniques to test for the presence of chemicals/molecules in our water
· To find other resources of information on water quality
ACTIVITIES
What you will do in lab today-
· Each pair of students will choose a sample of water to test
· Each pair of students will perform assays on their water sample to determine whether it contains iron, chlorine, ammonia, calcium or magnesium, nitrates and oxygen
· Each pair will report their results on their water sample
ASSESSMENTS
· Lab report (75): This is your first lab report of the semester – make sure you read the resources on writing a lab report and use the rubric – you can link to those here .
INTRODUCTION
Water is a chemical (H2O) that all life depends on. It makes up about 60% of the human body and our cells and organ systems are dependent on its unique characteristics. It is an important solvent, which means that many things dissolve into it (see below for a short list). For example, table salt easily dissolves in water. Also, water naturally has a pH of 7, but when other chemicals are present the pH can change. When pH decreases, we call that acidic.
As human population grow, so do their needs for, including their need for water. Many researchers believe that in the next 20 years over half the world’s population will have a shortage of potable or drinkable water. Despite the fact that 75% of the surface of earth is covered in water, 97% is salt water in the oceans and another 2% is frozen in glaciers, leaving us with less than 1% to use. Scientists hope to develop methods to increase potable water using desalination techniques, but in the meantime we have to take care not to contaminate or pollute our current drinkable water sources.
Toxins in the water are absorbed or consumed by primary producers and primary consumers – those “low”on the food chain – like bacteria, algae, protists and invertebrates. As toxins are transferred “up the food chain” or from one trophic level to the next, they are concentrated and become even more toxic to the organism. This phenomenon is called bioaccumulation or biologic magnification (Figure 1). This is one reason that humans need to be concerned about pollution in natural rivers and lakes in addition to our public drinking water.
Figure 1. Bioaccumulation or biomagnification. The accumulation of a chemical (in this case the + signs) accumulates as the trophic level increases from I to IV.
There can be tremendous variation from one water sample to the next. Here are some chemicals that can dissolve in water and how they affect its quality. Keep in mind that just because a chemical or ion is present, does not mean the water is dirty or non-potable.
1. Iron (Fe): This element can sometimes be found at high levels in well water because the iron in rocks dissolves into rainwater as it seeps deep into the earth. Iron is not unhealthy, but high levels can discolor the water making it appear reddish-brown – high iron can discolor food that is cooked in this water.
2. Chlorine (Cl): This is a powerful oxidizing chemical used to kill microorganisms in our drinking water, making it safer to drink. However, this means that significant levels of chlorine are found in most waterways.
3. Ammonia (NH4): Ammonia is used in fertilizer, animal feed production, and certain kinds of manufacturing. This chemical is very toxic and the human body will immediately convert it to urea for excretion. Simpler animals such as fish just release ammonia into the water where it becomes diluted. But in fish tanks with unchanged water, it can accumulate and kill the fish. High levels can turn up in our water supplies. It can also be an indication of fecal contamination.
4. Calcium and Magnesium (Ca and Mg): These presence of these minerals are what determine the “hardness” or “softness” of water. Low concentrations of calcium or magnesium are soft – high concentrations make the water hard. Soft water requires less soap to produce lather and generally has a lower pH, which can become corrosive if too acidic. Water around DC is hard and this can cause a build-up of calcium deposits in pipes and in bathtubs.
5. pH: This is the measure of how acidic or basic water can be. pH 7 is the neutral pH that is optimal for most living organisms. Low pH indicates acidic conditions. You may have heard about acid rain in the news. Plants and animals are sensitive to acid rain, as well as water that is very basic and has a high pH.
6. Nitrates and Nitrites: Living organisms require a source of nitrogen and they must have it in the form of nitrate. Nitrates are found in fertilizers and can accumulate, especially in water. Nitrates are a primary water pollutant because when it rains, the nitrates are washed out of yards, farms and fields into rivers, lakes, streams, and eventually the oceans. If there are high levels of nitrates in the water, algae will overgrow and use up all the oxygen, which depletes oxygen from fish and larger animals.
7. Oxygen (O): Living organisms including those that live in water require oxygen. Oxygen can dissolve in water at different concentrations depending on temperature and the presence of photosynthesizing organisms. There are currently large areas of the Chesapeake Bay that are oxygen-depleted as a result of pollution from nitrates and phosphates. Nothing can survive in these waters.
As you can see from the list above, water can vary dramatically from one source to the next. In addition to chemicals, living organisms can also be in drinking water. For example, bacteria are everywhere in our environment but most are not pathogenic (harmful). One class of bacteria that is considered a problem if it gets into our water systems is the Enterobacteria. This group includes E. coli, which can cause disease. Clean drinking water is treated and clear of these particular types of bacteria.
For more on the properties of water visit the corresponding chapter in the Concepts of Biology text:
For more information about water quality and testing visit this website.
· https://water.usgs.gov/edu/waterquality.html
To learn more about the Environmental Protection Agency and their policies on water quality in our area visit this website.
· https://www.epa.gov/standards-water-body-health/what-are-water-quality-standards
BASIC METHODS & EXPERIMENTS
The class will be divided into groups and each will analyze a water sample – groups will test the same water sample in a series of assays. First, make observations of the water sample, including its appearance and odor. Record these observations in your lab notebook! Based on these observations and what you know about the source of the water, make a hypothesis and prediction about the characteristics and/or quality of this water sample. Then each group will perform the assays on their water sample and record the results in their notebook.
Stations are set up for each test with positive and negative control solutions for:
1. iron
2. ammonia
3. magnesium/calcium
4. chlorine
Positive and negative controls are samples that are tested alongside your water sample – the scientist selects these because he/she knows that they contain or do not contain the substance being tested for. The positive control has the substance being tested and the negative control does not. You must compare the results for your sample with the positive and negative controls at each station to determine if that substance is present in your water sample. This means you will set up three tubes at each station: 1) your water sample, 2) a positive control and 3) a negative control. The reagents and directions for each test will be organized at stations around the room.
In addition to these qualitative assays that you will perform, you will:
1. examine the bacterial content of your water sample
2. determine the amount of dissolved oxygen in your water sample
3. determine the pH of your water sample
4. determine the level of nitrates in your water sample
5. determine whether D.C. water tests positive for lead
DETAILED PROTOCOLS & PROCEDURES
PROCEDURE
Start by adding 0.5 mls of your water sample to 6 microcentrifuge tubes. Label the tubes for each of the tests.
STATION TESTING FOR IRON
Positive Control = Ferrous Chloride; Negative Control = Distilled Water
1. Using a clean pipet, add 5 drops of sulfuric acid to each sample in the microcentrifuge tubes. Close the lid and shake well.
2. Using the scoop provided, place a very small scoop of ammonium thiocyanate crystals into the microcentrifuge tubes. Close the lid on the tube and shake well
3. The formation of an orange or dark purple solution is a positive indicator for the presence of iron.
4. Record your results for this test in the appropriate space in Table 1. writing either (+) to indicate the presence of iron or (-) to indicate the absence of iron.
Station Testing for Chlorine
Positive Control = 1% Chlorine Bleach; Negative Control = Distilled Water
1. Using a clean pipet, add 3 drops of O-Tolidine to the samples in the microcentrifuge tubes. Close the lid on the tube and shake well.
2. The formation of a reddish-brown color is a positive indicator for the presence of chloride. Light tan or yellow is negative.
3. Record your results for this test in the appropriate space in Table 1. writing either (+) to indicate the presence of chlorine or (-) to indicate the absence of chlorine.
Station Testing for Ammonia
Positive Control = Ammonium Chloride; Negative Control = Distilled Water
1. Pipette 0.5mL of your water sample into a microfuge tube.
2. Add 2 drops of Ammonia Test Solution #1.
3. Add 2 drops of Ammonia Solution #2.
4. Close the lid and shake the tube well for 5 seconds. Wait 5 minutes.
5. Determine the amount of Ammonia in your sample by comparing it to the color chart. Record the amount in your lab notebook.
Station testing for Calcium and Magnesium (Hard and Soft Water Station)
Positive Control = Magnesium Chloride & Calcium Chloride; Negative Control = Distilled Water
1. Using a clean pipet, add 5 drops of ammonium hydroxide to the samples in the microcentrifuge tubes. Close the lid on the tube and shake well.
2. Using a clean pipet, add 2 drops of the indicator solution, Eriochrome Black T, to the samples in the microcentrifuge tubes. Close the lid and shake well. The solution should be pink at this point.
3. Using the pipet provided, add 5 drops of EDTA to the tube. Close the lid on the tube and shake the tube well.
4. If color changes to blue then the water is soft (low level of metal ions). If the water stays pink then it is hard and has high levels of metal ions.
5. Record your results for this test in the appropriate space in Table 1. writing either (+/pink) to indicate high levels of metal ions or (-/blue) to indicate low levels of metal ions.
Station testing pH and Presence of Nitrates
There are no positive or negative controls for this station.
Determining pH
1. Pipette 0.5mL of your water sample into a microfuge tube.
2. Add 2 drops of universal indicator to the sample.
3. Close the lid and shake the tube well.
4. Compare the pH of your sample by comparing it to the color chart. Record the pH in your lab notebook.
Determining levels of Nitrates
1. Pipette 0.5mL of your water sample into a microfuge tube.
2. Add 2 drops Nitrate Test Solution Bottle #1 of the sample in the microfuge tube.
3. Close the tube and invert several times to mix.
4. Vigorously shake Nitrate test tube solution #2 for 30 seconds
5. Add 2 drops Nitrate test solution bottle #2 to the sample microfuge tube.
6. Close the tube and vigorously shake for 1 minute. Wait 5 minutes.
7. Determine the amount of Ammonia in your sample by comparing it to the color chart. Record the amount in your lab notebook.
Station testing for Amount of Dissolved Oxygen
You will use the Winkler method and titration to determine the amount of dissolved oxygen in the water sample. Ultimately, the volume of thiosulfate you add is directly correlated to the amount of DO in your water sample and this value is converted to parts per million (ppm) oxygen.
The instructor will do steps 1-5 to preserve the dissolved oxygen. We will use 50 ml water samples.
1. Add 166 microliters of MnSO4 solution.
2. Add 166 microliters of alkali-iodide-azide solution.
3. Invert several times to mix, thereby allowing a brown precipitate to form.
4. Let the precipitate settle to at least half of the bottle/test tube volume.
5. Observe the brown precipitate!! The TA will micropipet 166 microliters sulfuric acid into the sample tube. Invert the tube to dissolve the brown precipitate.
6. Pour the 50 ml sample into an Erlenmeyer flask, set it on a stir plate and set the stir bar spinning.
7. Use a P1000 pipet to slowly add in 1ml of sodium thiosulfate.
8. Add 20 drops of starch solution and mix. The sample should turn to a darker brown color.
9. Continue adding sodium thiosulfate in 100µL (0.1ml) amounts. You must continue until the brown does not come back. Keep track of the amount added. Use the Table in the worksheet to record amounts.
10. Calculate the total amount of sodium thiosulfate added in steps 7 and 9. For the 50 mls sample we are working with, 1ml of added sodium thiosulfate is equal to 6 ppm or 6mg/L oxygen. So multiply the number of mls of sodium thiosulfate added by 6 to get the total ppm. Record the total ppm of oxygen in your lab notebook.
Coliscan is a commercial system that tests for the presence of Enterobacteriae in water samples. The patented media includes two color producing chemicals, one for the detection of the enzyme glucuronidase (produced by E. coli strains but not by general coliforms) and one for the detection of galactosidase (produced by all coliforms, including E. coli). Thus, if coliforms are present a pink pigment will color the colonies. If E. coli (a specific type of coliform) is present, there will be bluish-purple colored colonies. The Coliscan system was set up by the instructor prior to class. Observe the plate and note the numbers/colors of the resulting colonies in your lab notebook.
An example of a Coliscan plate. Each colored dot represents a bacteria that was present in the water sample. Bluish/purplish colonies indicate E. coli.
RESULTS & CONCLUSIONS
Record the following in your lab notebook:
1. The source of your water sample
2. Your initial observations of your water sample and hypothesis and prediction for what it might contain.
3. Below is a table that you can copy into your lab notebook and use to record the data about your own water sample in addition to the samples from the other groups.
4. Take a picture (carefully, with your phone) of the agar plate showing bacterial load in your own water sample.
5. Record any deviations, mistakes or surprises about your methods and results. These notes will help you write a coherent and interesting lab report later.
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Chemical Tested For |
Positive Control (describe assay results) |
Negative Control (describe assay results) |
Sample 1______ (+ or -) |
Sample 2______ (+ or -) |
Sample 3______ (+ or -) |
Sample 4______ (+ or -) |
Sample 5______ (+ or -)
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Sample 6______ (+ or -)
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Iron |
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Chlorine |
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Ammonia |
N/A |
N/A |
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Calcium & Magnesium |
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pH |
N/A |
N/A |
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Nitrates (ppm) |
N/A |
N/A |
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ppm O2 |
N/A |
N/A |
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# E. coli(dark blue/purple colonies) |
N/A |
N/A |
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# Other coliforms (pink/red colonies) |
N/A |
N/A |
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# of other bacteria (green colonies) |
N/A |
N/A |
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Table 1: Water Sample Results
To keep track of dissolved oxygen, use a table similar to below:
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Total Number |
Total Volume (in mL) |
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# of 1 mL aliquots of Na Thiosulfate added |
___________ |
___________ |
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# of 100 uL aliquots of Na Thiosulfate added |
___________ |
___________ |
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Total volume of Na Thiosulfate in mL |
N/A |
___________ |
Each mL of Na Thiosulfate added equals 6 parts per million (ppm) or 6 mg/L of dissolved oxygen. Multiply the total volume of Na Thiosulfate in mL to calcualte the amount of dissolved oxygen.
ASSIGNMENT – LAB REPORT
WATER QUALITY LAB REPORT (75 POINTS)
Each of the bold parts below are used as section headings in your lab report. Your report should be in a 12-point font, double-spaced and no longer than five pages (including the tables). Consult the lab report rubric in addition to the details below.
Title (make it specific to the topic and your water sample)
Introduction: In 1 paragraph include the following:
· References that provide at least three pieces of background information on how the quality of water is tested.
· State the hypothesis and prediction about your particular water sample. Make sure your hypothesis has an explicit justification and is not just a guess or your opinion. Use sources to convince your reader!
· One or two sentences to connect your background information with the importance of your hypothesis and your prediction for the experiment.
Materials and Methods
State which assays you performed and the positive and negative controls for each. Make sure you use past tense and do not include a materials list or any reference to containers!
Results
Include a description of the water sample and the source of the water. Summarize the results of your water sample at every station, citing your data tables (1-3). Include the following data tables in your results section.
Table 1: Water Sample Results
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Chemical |
Positive Control |
Negative Control |
Sample 1_____ |
2 _____ |
3 _____ |
4 _____ |
5 _____ |
6 _____ |
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Iron |
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Chlorine |
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Ammonia |
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Calcium Magnesium |
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pH |
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Nitrites (ppm) |
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Nitrates (ppm) |
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ppm O2 |
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Table 2: Presence of Bacteria
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Sample: _______ |
# colonies/ plate |
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# E. coli (dark blue/ purple colonies) |
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# Other colifoms (pink/ red colonies) |
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# of other bacteria (green colonies) |
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Table 3: Dissolved Oxygen Calculations
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Sample: __________ |
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# 1mL aliquots of thiosulfate before starch |
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# 100µL aliquots of thiosulfate after starch |
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Total volume (in mL) thiosulfate ppm oxygen |
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Discussion
· Summarize ways in which the water can be affected by the environment. How did this influence your hypothesis and prediction?
· Describe how your results supported or refuted your hypothesis and your prediction. You may also need to explain why results were inconclusive. Make sure to include the outcome of positive and negative controls if results are inconclusive.
· Describe any problems os issues with observing and quantifying the assays, oxygen titration, and coliform bacteria colony growth.
· If water is clear and smells fresh, can you consider it safe to drink? Give two reasons why or why not.
· Look up the Clean Water Act that congress established in 1972. Is it still important to current day?
· In a couple of sentences, write a conclusion about the importance of safe drinking water in terms of globalization and the biological significance to growing human populations.
References (You should use at least 3 outside references. Cite these in the text as you reference them!)
(rubric)
THE LAB REPORT AND SCIENTIFIC WRITING
Scientists report the findings of their experimental research in a very precise manner that reflects the scientific method. Usually a paper that is submitted to a science journal is the culmination of many many experiments. But whether it is a journal article or the lab report on the single experiment you have completed today, the overall format is the same. It is essentially the written story about an experiment/s.
The writing is broken up into five distinctly labeled sections: Introduction, Materials and Methods, Results, Discussion and References.
1. Introduction – This section introduces relevant background information helpful to understanding the experiment being described. It should include the purpose of the experiment, why your hypothesis and prediction are important, and how your hypothesis relates to the background information. This section is short, usually only one or two paragraphs. You will use in-text citations in this section!
2. Materials and Methods – The procedures and materials used are described in a few paragraphs. It is always in past tense. You need not include a materials list!
3. Results – Charts, graphs, tables, and drawings are the best way to show any data. Make sure each has a number, a title, and the parts are labeled. There should also be a description for each figure. Observations belong in this section. However, no interpretations or explanations of the data and observations are included, just the plain facts here.
4. Discussion – The Discussion summarizes and connects all the parts together. It is always important to describe how the information in the Results sections either supports or does not support what was predicted in the Introduction. The results are interpreted and related to both the hypothesis and background information. Any problems with the experiments are usually mentioned. Then an alternative hypothesis or new predictions can be stated and the importance of the findings is stressed. Finally, end with any conclusions you can draw from your data and the class data (not personal opinions). This section demonstrates how well you understood the lab.
5. References – Cite the sources of any background material, including your lab manual and web sites. Three is the minimum! Never use any quotes in a report or worksheet!!
TIPS ON WRITING A FANTASTIC LAB REPORT!
1. This is a link to a fantastic resource that will get your scientific writing off to a great start. The guidelines in this resource are consistent with the expectations of this course and our department.
2. It might also be useful to read some great lab reports and some not so great lab reports – you can find links to those here.
COMMON PROBLEMS WITH LAB REPORTS (READ THESE!)
This is a link to a fantastic resource that will get your scientific writing off to a great start. The guidelines in this resource are consistent with the expectations of this course and our department.
General:
1. Don’t forget your name, TA name, section and date 2. Use standard borders and 12point fonts 3. Don’t use the title from the manual…it is not specific enough 4. Make the title concise and to the point
Introduction:
1. Reference sources for background information 2. Introduction is not a summary of how the experiment was done 3. Include information that is needed to understand the meaning of the next three sections 4. Don’t explain concepts that are already understood by scientists such as why controls are needed. 5. End your introduction with the hypothesis and If…then prediction (good way to transition into the next section). Include the information which led you to formulate this hypothesis and prediction.
Materials and Methods:
1. Don’t divide into two sections. And don’t provide a list of materials used! 2. Write brief summary of materials and methods and site the manual instead of writing all the details. 3. Make sure any changes or additions to manual instructions are included. 4. Be as concise as possible without sacrificing clarity. 5. Make sure to use the past tense.
Results:
1. State results in paragraph form but DON’T interpret them. 2. Describe trends in data, factually. (Positive correlation between protein concentration and % absorbance) 3. Explain the reasoning behind more complex calculations. 4. All tables and figures must be numbered. Tables are numbered sequentially in the order in which they are referred to in the Results section. Any data presented that is not in a table is considered to be a figure. Figures are also labeled sequentially in the order in which they are referred to in the Results section.
Tables and Graphs:
1. Data may be presented in table form. Each column and row should be appropriately labeled, and the table itself should have a title. Be sure to include units of measurement in the labels where appropriate. 2. Graphs are used primarily to reveal trends that would not be obvious in tabular form. Each X and Y axis must be labeled appropriately and the units should be given. When plotting data on a graph, indicate each data point clearly with a symbol such as a filled-in or open circle or square. If there are two or more plots on a single graph, use two different symbols to indicate the data points and include a clearly labeled legend showing which symbol represents which plot. 3. Other data, such as micrographs, pictures or drawings must also be labeled. They are considered to be figures, as are graphs, and must have a legend briefly describing the figure, including the organism, scale and any other appropriate information.
Discussion:
1. Begin with brief summary of background information and purpose of the experiments. 2. Include a brief (1-2 sentences) summary of results and hypothesis. 3. Did data support hypothesis? Explain why or why not. 4. Be thorough when explaining sources of error (human and equipment). How did they affect the experiments and how can they be fixed or minimized? 5. Interpret meaning and importance of results. How results could be used by others or in future experiments (what have they contributed to science). Explain how experiments apply to “real world” issues, problems, or concerns
References:
1. Cite as shown in introduction. 2. Don’t use direct quotes. 3. Don’t footnote. Include a references section at the end of the lab report or worksheet.
HOW TO CITE THIS LAB MANUAL IN YOUR LAB REPORT
Below is the proper citation for the current version of your Biology 100 lab manual. Please use it in all your assignments that cite the manual.
Pedersen-Shear, A., M. Bentley, S. Frances-Knight, N. Zeller and K. Walters-Conte. (2016). BIO-100: Great Experiments in Biology, Lab Manual. Retrieved from http://bio100labmanual.openbooks.wpengine.com/
RUBRIC FOR YOUR LAB REPORTS
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Introduction ( /15 points) |
(~0 points) |
(~1 point) |
(~2 points) |
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Criteria |
Weak |
Satisfactory |
Strong |
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Purpose/goal of experiments is clearly stated |
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Motivation and/or importance of the study clearly stated |
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Background information appropriate to the study is included |
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Resources are appropriately cited |
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A logical and clear hypothesis and prediction are stated |
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Information flows logically from general to specific |
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The approach to carrying out the study is briefly stated |
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Results Text ( /15 points) |
(~1 points) |
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(~3 points) |
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Criteria |
Weak |
Satisfactory |
Strong |
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Reader can understand what particular experiments show |
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Data trends and patterns noted and described accurately |
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Appropriate figures/tables referenced in the text |
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Reader can envision data in figures and tables from reading the results text alone |
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Results presented in sequence that allows the reader to understand how the experiments relate to one another |
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Materials and Methods ( /5 points) |
(~0 points) |
(~0.5 points) |
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Criteria |
Weak |
Satisfactory |
Strong |
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Presented in logical order |
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Final working concentrations of chemicals/solutions stated |
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Procedures accurately and succinctly described |
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Chemical and scientific names accurately and consistently used |
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In past tense with no reference to labeling or containers |
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Could a reader, generally familiar with scientific procedures and process, understand the basis for the experiments performed and data produced? |
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Results Figures and Tables ( /15 points) |
(~0 points) |
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Criteria |
Weak |
Satisfactory |
Strong |
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Appropriate format/graph type for data chosen |
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Figures and tables follow a consistent format |
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Legend/caption present and in consistent location |
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Legend information complete, accurate and specific |
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All axes, columns, etc. are labeled properly |
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Figure/table understood without referring to results text section |
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Proper numbering sequence of data figures and tables |
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Figures and tables are aesthetically acceptable (reasonable size, not separated by page breaks, text alignment consistent) |
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References ( /5 points) |
(~0 points) |
(~0.5 points) |
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Criteria |
Weak |
Satisfactory |
Strong |
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Format of in‐text references is correct |
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References are papers that relate to the topic of this study |
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The information used from each reference cited adds value |
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Format of references at the end of the text is correct |
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Number of references appropriate and sufficient |
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General Paper Mechanics ( /10 points) |
(~0 points) |
(~0.5 points) |
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Criteria |
Weak |
Satisfactory |
Strong |
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Spelling |
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Grammar |
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Information flows logically and smoothly through paragraphs |
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Paragraphs adhere to a topic |
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Sentence structure is concise and clear with good transitions |
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Terminology/jargon is used consistently and correctly |
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Personal opinion is avoided; not cited as fact when used |
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Excessive detail/brevity is avoided |
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Includes a specific title and students name |
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