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11 Aug 2019

1. Dr. X wanted to see if fermenting grapes would produce more achohol if more sugar was added than he normally added (50 g). His experiment had 2 groups: group 1 had 50 g of sugar added to 2 1b. of grapes and 1 liter of water. Group 2 had 100 g of sugar added to 1 1b. of grapes and 1 liter of water.

A. what is the experimental group? -----
B. Explain your answer.
c. what is the control group?
d. explain your answer.

2. Brine shrimp or sea monkeys normally live at pH 8. you want to test the effect of pH on the ability of brine shrimp to survive at a lower pH (pH 4, 5 6 and 7).

a. what would be the experimental groups?--------'
b. explain your answer.
c. what would be the appropriate control group?
d. explain your answer.

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11 Aug 2019

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For Discussion (2-3 paragraphs) â¢ What was the concentration of your pre-diluted NPs? â¢ Determine the effect of NPs on brine shrimp lethality. â¢ Determine the effect of NPs on cell viability. â¢ What is (are) the control(s) in this experiment? What is its purpose? â¢ Based on your data in this lab, what is a safe concentration for brine shrimp Threshold Level of Toxicity (TTL)? â¢ Often indicator species are used to study the overall health of an ecosystem. If you were to study an ecosystem containing brine shrimp, would you use it as an indicator species? Why or Why not? Explain your reasoning. â¢ What possible sources of error were present in this experiment?

Introduction

LD50 is a commonly used term to describe acute toxicity: LD stands for Lethal Dose and 50 is used because the dose could kill 50% of the animals exposed to the chemical. For LD50 tests, the animals are given specific amounts of the chemical, orally, by injection or dermally. The value is then reported along with the method of administration and the type of animal used in the test. LD50 is typically reported in milligrams or grams per kilogram of body weight of the animal. The lower the LD50 is, the higher the toxicity of the chemical would be. This information is always provided in Safety Data Sheet (SDS) of chemicals in the âtoxicity dataâ category. A similar measure, the LC50 is usually used to describe the ecological hazards of chemicals. LC stands for lethal concentration that could kill 50% of the animals after 24-96 hours. This value is always provided in Safety Data Sheet (SDS) of chemicals in the âEcological Dataâ category. In this lab, you will be measuring LC50 or LD50 of iron oxide nanoparticles using brine shrimp. Iron oxide nanoparticles are magnetic. Application of magnetic nanoparticles ranges from magnetic storage media to medical industry where they can be used for targeted drug delivery or as contrast agents in MRI. Thus, measuring environmental and biological toxicity of these nanoparticles is crucial. To test toxicity of any such reagents on small organisms, Daphnia and brine shrimps (also known as Artemia) can be used. Artimia are found in salt lakes around the world. They are important live food for aquarium fish. In this lab, you will use brine shrimp to measure toxicity of the nanoparticles. Trypan blue is a stain used to assess cell viability. It is able to diffuse through cell membranes of dead cells but is unable to permeate live cells. This stain can be used to assess cell survival 1 Fall 2017 following exposure to chemicals, nanoparticles and experimental drugs. The ratio of live: dead cells can be used as a preliminary assessment of cytotoxicity and translated to tissue viability.

Methods Part A: Serial Dilution of NP Solution 1) Label 5 test tubes as 0, 1:1, 1:10, 1:100, 1:1000 2) Add 2mL of seawater to tube 0 3) Add 2mL of specific group stock solution to tube 1:1 and add 2mL of seawater to it. 4) Transfer 1mL of stock solution to tube 1:10 and add 9mL of seawater to this tube 5) Transfer 1mL of 1:10 solution to the tube labeled as 1:100 and add 9mL of seawater to â¨this tube 6) Transfer 1mL of 1:100 solution to the test tube labeled as 1:1000 and add 9mL of â¨seawater to this tube Part B: Preparation of test environments 1) Label 5 wells of 6-well plate as control (0), 1:1, 1:10, 1:100 and 1:1000 2) Label 3 petri dishes as: control (0), 1:1 and 1:1000 â¨NOTE: Be sure to include your initials on all plates 3) Transfer 2mL of each solution from the corresponding test tube to the respective wells on your six-well plate and each petri dish Part C: Preparing specimens for the Trypan blue cell viability test 1) Transfer 2 brine shrimp into each of the pre-labeled petri dishes containing the solution of NPs and seawater (0, 1:1 & 1:1000) and set a timer for 30 minutes. 2) Transfer a single brine shrimp into the unlabeled petri dish with a drop (~50ul) of seawater and use the dissecting microscope to make observations related to the behavior and morphology of untreated brine shrimp.â¨Make observations in lab notebook of all samples Part D: Preparation of specimens for the toxicity assay 1) 2) 3) 4) 5) Add 10 brine shrimp into each well in the 6 -well plate Make observations in lab notebook Using a dissecting microscope (if needed), count the number of brine shrimp in each well (alive shrimps).â¨Record your data in your notebook using the graphic below as a template. Close the plate and store it for next observation.â¨Observations must be done within 24h-96hâ¨Note: There will be times allocated and a sign up paper for observations Part E: Trypan blue viability test 2 Fall 2017 1) Label 3 glass slides to match the petri dishes prepared in Part C. 2) Transfer 1 brine shrimp from each petri dish onto the corresponding slide. â¨Make observations in your lab notebook 3) Add a drop (~60 Î¼l) of PBS on top of each specimen 4) Add 60 Î¼l of Trypan blue to your shrimp and incubate for 1-2 minutes at room â¨temperature. 5) Wick away excess liquid 6) Add 100 Î¼l PBS to sample and observe (while alive!), looking for blue cells, using the â¨dissecting scope â¨Make observations in your lab notebook 7) Label 3 microfuge tubes to align with each sample. 8) Transfer each brine shrimp from the slide into the corresponding microfuge tube 9) Add 50 Î¼l of PBS to the tube and use the mechanical disruption tool to homogenize the â¨sample for ~2 minutes until a cell suspension has been generated. 10) Load 30-50 Î¼l of your sample onto a new glass slide and add a cover slip. 11)Observe the sample on high power and use the scale below to qualitatively assess cytotoxicityâ 0-no blue 1-some blue 2-fairly blue 3-totally blue.

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Experimental Review

Below are five hypothetical experimental designs. Your job as a reviewer is to pick one of the experiments and write a detailed summary of what is wrong with the proposed study.

Your review should use vocabulary words that have been discussed in class and address the following questions.

Is the study feasible? Will it actually be possible to conduct the proposed study in a controlled manner that produces meaningful data? If there is more than one variable changing simultaneously, no controls, or no replication, then the study is unlikely to produce meaningful data. If there is no way to monitor an experiment in the environment, or no way to conduct the experiment in a controlled environment, like a laboratory, then the experiment is unlikely feasible.

Can you tell from the text if the experiment is designed to be conducted by performing treatments, or by analyzing observational data about a species and its habitat?

Is there a testable hypothesis? Is the study set up so that it will be possible to measure the effects in a quantifiable way? What will be measured? Is there a control group and a treatment group? Or, an independent variable like temperature, and a dependent variable like germination time?

Could you graph the results of this experiment? If so, how would you do it? A column chart to show treatments and controls? A scatter plot or a line graph to look at how changes in the independent variable affect the dependent variable?

Create a post on the discussion board by clicking on "Click to Launch" above.

1. Experiments to Review

1. Climate Change Effects on Marine Iguana Mortality

Hypothesis: If the water temperature in the Galapagos Islands changed from being hot (around 77 degrees) to cold (under 70 degrees), then the mortality rate of Marine Iguanas will increase.

Variables: The independent variable is the temperature of the water that the Marine Iguanas live in. The dependent variable is the mortality rate of the Marine Iguanas. The mortality rate of the Marine Iguanas at the average water temperatures of their habitat.

Experimental Treatments: Iguana Group 1 will be water temperatures and marine iguana mortality rates before 1998. Iguana Group 2 will be water temperatures marine iguana mortality rates after 1998

Prediction:By comparting the average water temperatures with the mortality rates every year before the sudden drop in the iguana population and after a trend will be able to be seen. If the water temperature is truly to blame for the mortality rate increase the water temperature will decrease as the population of Iguanas decreases. The control group is intended to show the average number of Iguanas that die due to other causes.

2. Volcanic Ash Impacts Marine Iguana Longevity

Hypothesis: There are several volcanoes between both the Isabela, and Femandina islands which make this the obvious culprit for the extreme rise in mortality in Marine Iguanas.

Variables: The independent variable is the number of volcanic eruptions per year. The dependent variables are the change in pH of the water in the marine iguanasâ habitat, and the mortality rate of the iguanas.

Experimental Treatments: Iguana Group 1 will live on an island with multiple volcanic eruptions each year. Iguana Group 2 will live on an island with no volcanic eruptions.

Prediction: The group of iguanas living on the island with no eruptions will have longer life spans because the pH of the water will not be altered by the large number of volcanic eruptions.

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You and your hypothetical lab partner each develop an experimental plan to test this hypothesis. The two experimental plans are presented below. Evaluate each plan for its pros and cons in the experimental design using the bullet points below. Both experimental plans have some good elements and areas that could be improved.

Experiment A) Mature 2 inch sections (sprigs) of Elodea will be used to measure oxygen production under different light conditions. 5 sprigs will be tested. One will be kept in the dark (no light). The others will be exposed to a light filtered with cellophane. One sprigâs light will be filtered through clear cellophane to receive all wavelengths of light, and the other sprigsâ light will be filtered through red, blue, and green cellophane, respectively. The no-light plant will be kept in a dark cabinet, while the other plants will be together on a counter. Each plant sprig will be placed in equal sized upside-down test tubes in 100ml of tap water so that oxygen bubbles produced by the plant will be trapped at the top of the test tube. A ruler will be used to measure the length of the oxygen pocket collected at the top of the test tube. The experiment will continue until one sprig produces 5 cm of oxygen in the tube.

Experiment B) 3 cm sections (sprigs) of Elodea of various ages will be used to measure oxygen production under different light conditions. 15 sprigs will be divided into 3 groups of 5 sprigs each. One group will receive natural sunlight, one group will be exposed to a blue light bulb under a box to block natural light, and the final group will be exposed to a red light bulb under a similar box to block natural light. Each group of plant sprigs will be trapped under a clear plastic funnel that is covered by an upside-down test tube to collect oxygen bubbles produced by all the sprigs in the group. All groups will be in 100ml of water of the same temperature and pH. The experiment will continue for 24 hours, and then the volume of oxygen gas produced will be measured by multiplying the length of the oxygen pocket by the circle described by the test tube, ?r². The average volume produced by all groups will be graphed and compared to determine which condition produced the most total volume of oxygen.

Consider the following questions to evaluate experimental plans A and B (DO NOT ANSWER THESE QESTIONS)

What is the independent variable suggested by the hypothesis? Do both plans test the independent variable equally well?

What is the dependent variable in each experiment? Which experimentâs dependent variable best matches the prediction in the hypothesis?

What are the experimental and control groups for each experiment? Which experiment gives the best opportunity to assess the effect of the independent variable?

How does each experiment account for standardized variables? Which experiment do you think is the better controlled experiment?

How is sample size addressed in each experiment? Which would you find to be more convincing?

Does either experiment describe any statistical analyses to be performed on the data?

ANSWER THIS QUESTION: BELOW

Considering your analysis of each of the questions above,write an improved experimental design to test the hypothesis âIf blue light is more effective at promoting photosynthesis, then more oxygen gas (O₂) will be produced when plants are exposed to blue-filtered light compared to other wavelengths.âYou may use elements of the above experiments