Kristen Swafford, Maddy Hall, Olivia Henrion, and Julia Ellen

Collaborators and Roles

AP Environmental Science

Kara Norris

2 March 2017

LD50 Lab

Introduction

In this experiment, we conducted a dose response bioassay in order to find the LD50 of radish seeds in

different water dilutions. A dose response bioassay is an experiment conducted by taking a population of test

organisms and exposing them to different amounts of a test substance. The tested organisms responses are then

recorded and analyzed. LD50 occurs when a dose of a drug or pollutant kills half of the test population members.

LD50s are often tested on animals, such as mice, and then applied to humans. The use of LD50s are very helpful

in the medical field because they can help doctors to create the amount of medicine a person should take (dose)

as well as how much pollution is dangerous. (Thorpe)

The LD50 of salt for a radish is important because farmers use them to renew their soil. This is because

radishes reduce the need for fertilizer and retain moisture and reduce erosion (Hannah). These aspects help

save money and time for farmers when their soil degrades. One way soil degrades is high concentrations of salt.

This degraded soil has caused $27.3 billion annually in lost crop production (Zielinski). Salt is added to soil

through irrigated water and road salt. Irrigated water contains dissolved salts that are left behind when water

evaporates (Zielinski). Using irrigation is essential in areas where there is a lack of water, but it can lead to

further crop loss. Road salts also contribute to high concentrations of salt. This is because road salts dissolve in

sodium and chloride ions that are then washed away into waterways. The waterways transport and disperse the

salt into different areas, which contributes to salt buildup (Stromberg). However, radishes are a low salt tolerant
plant (Badurek). This can be problematic for the farmers with high salt concentrations in their soil, as the

radishes may not grow. By finding the LD50 , it would be beneficial to the farmers, who use radishes to renew

their soil.

According to the United Nations University, the Earth is losing valuable soil each day due to salt

induced degradation (Qktheme). With this being the case, valuable farm land is slowly running out. If we have

lower amounts of soil that are good for farming, it reduces the amount of produce we can grow to feed the

people. Some plants can withhold higher levels of salt in the soil and still prosper to their fullest extent, whereas

others can not stand the higher levels of salt. Eventually, the plants will adapt to the salt in soil, some more than

others. Although that might not always be a good thing in the end for the consumers, we should try to do what

we can to make sure that the salt concentration does not exceed the LD50.

Purpose

To determine the effect of differing salt concentrations on the germination and radicle root growth of

radish plants.

Hypothesis

If the salt concentration increases, then the radicle root length will decrease as compared to the 0%

concentration control, and there will be a 50% mortality rate when the salt concentration is at 25%.

Materials & methods

1. Using 20 ml of the original concentration of salt perform a serial dilution to create five different salt

concentrations, plus a sixth with distilled water for the control.

2. Count out 60 seeds, 10 for each water sample.

3. Put 10 seeds into a folded paper towel, space the seeds out evenly on the paper towel so that they do not

touch each other.

 4. Wet the paper towel/seeds and seal in a baggie, be sure to label the bags with the salt concentration for

that sample.

5. After 4-5 days examine the seeds, counting the number of each that germinated. You will measure the

response of the radish seeds at various salt concentrations.

6. Remove the lid of the control dish. Count the number of seeds that germinated. Calculate the

percentage of seeds that germinated and record in Table 2.

Note: if fewer than 80% of the seeds in this control sample

germinate, this indicates a problem with the experiment.

7. Measure the length of the radical for each of the

germinating lettuce seeds to the nearest millimeter (mm). Look carefully at each sprout to make sure

you are measuring just the root, not the shoot as well. In the picture below, you would measure just the

part between the two arrows, not the shoot and cotyledons to the left.

8. Repeat steps 6-7 for each petri dish.

9. For each treatment, calculate the mean radical length for each salt solution. Add the total radical lengths

for each salt solution and divide by the total number of seeds that germinated. Do not include data from

seeds that did not germinate. Record data in column labeled, Mean Radicle Length (mm).

Data and Analysis

# of Radical Root Length (mm)

seeds
Concentration germi Mean Differenc
of Salt nated Radicl e in
(out of 1 2 3 4 5 6 7 8 9 10 e Radicle
10) Length Length
(mm)

Control 0% 10 40 70 35 55 55 90 80 60 70 60 61.5 0

6.25% 10 80 75 30 50 75 30 55 70 55 35 55.5 6

12.5% 10 37 40 80 100 70 90 90 25 65 80 67.7 -6.2

25% 8 50 55 19 15 7 30 30 22 0 0 22.8 38.7

50% 0 0 0 0 0 0 0 0 0 0 0 0 61.5

100% 0 0 0 0 0 0 0 0 0 0 0 0 61.5

Table 1: Number of Radish Seeds Ungerminated

Fig 1: Mean Radical Root Length

Great example!!Great example!!

Fig 2: Percent of Ungerminated Seeds at different salt dilutions

 While conducting this experiment, we found that the LD50 of
our data was approximately 27%. Three of

the different concentrations had all of the seeds germinate while two had no seeds germinate. Unexpectedly, the

seeds with the 12.5 salt concentration had a longer mean radicle length than the control concentration. The

threshold of this experiment was also at the 12.5% concentration. This is because past this concentration level,

seeds stopped germinating. Our experiment most likely differs from a real life scenario because the 12.5%

concentration did better than the control and this would most likely not occur in a normal scenario.

Conclusion

Through this lab, our hypothesis was disproved. Our hypothesis was disproved because the radicle root

growth increased at a concentration of 12.5% and the LD50 was 27%. We had hypothesized that the radicle root

growth was going to decrease and that the LD50 would be at 25%. These results were off from what we

originally expected. Also, we initially believed that the control group would have the longest radical length. Our

experiment results showed that the 0%, 6.25%, and 12.5% concentrations all had every seed germinate with the

25% concentration having 8 seeds germinate. The 12.5% concentration also had a mean radicle length higher

than the control group. The reason behind these results could be because the seeds were more concentrated in

one area and not spread through the whole dish. Because the seeds were spread out, the seeds would be exposed

to less salt because they were all obtaining it from the same small area in the dish. It is also very possible that

these results suggest that radishes can more easily adapt to medium levels of salt concentration and it does not

affect them as much until it reaches higher concentrations.

While the methods used limited the amount of error, there is still the possibility of error. The

measurement of the water and radicle roots could have been wrong. Having different amounts of water in the

container could have allowed for more growth in some of the containers and less in others. Also the radicle

roots became tangled if they grew too close to one another. This makes it harder to ensure that the

measurements of the root are correct since part of the root could have broken off. That can be avoided by

double-checking the measurement to make sure the measurement is correct.

 Our results accurately represented the purpose of this lab. The purpose was to be able to determine the

effects of differing salt concentrations on the radical root growth of radish seeds. The higher the salt

concentration, the smaller the radical root growth. The lower the salt concentration, the longer the radical root

growth. When we collected our data, the smaller radical root lengths were a result of the higher salt

concentrations and vise versa.

When we compared our final results with those of the class, we came to the conclusion that they are not

similar nor the same. The LD50 results range from 6.25% to 38% . There are a few cases where some LD50s are

similar, but overall, every group's results vary widely. This may be due to the fact sense that some people might

not have measured the radical root length correctly. Incorrect water and salt ratios might also be a reason why

the radical root lengths were not near the same. If a group incorrectly measured the amount of salt in each petri

dish of water, there are either going to be more germinated seeds (due to less salt concentration), or less

germination (due to too much salt concentration).

Scientists are utilizing similar LD50 experiments to conduct research that is important to agriculture and

other industries. For example, an experiment conducted by the University of Zurich suggests that the use of

livestock medicines can alter beneficial organisms that recycle and decompose dung. Scientists are conducting

LD50 experiments which test the amount of flies or maggots that die due to the use of Ivermectin, a livestock

medicine. This will help scientists to decide whether or not the medicine should be used (New Approach for

Environmental Test on Livestock Drugs). The medicine is given to livestock and excreted through their dung.

The organisms that break down and decompose the dung are at risk due to the Ivermectin. The results of this

experiment show that as the concentration of Ivermectin in the dung increased, so did the amount of organisms

affected (New Approach for Environmental Test on Livestock Drugs). These results are similar to the radish

salt concentration lab because as the salt concentration increased so did the amount of ungerminated radish

seeds. This experiment relates to this lab because the lab focused on the LD50 of the radish seeds when put into

different salt concentrations. However, in the experiment of the decomposing dung the scientists measured the

effects the medicine had on surrounding organisms. Although these experiments are measuring two different
types of variables having to do with LD50, they both are working on minimizing the risks of substances in

nature.

Some further questions for the study could include the following; What other plants could this

experiment have been conducted on? How would those results had differed? If we had left the radish seeds in

the petri dishes for a longer period of time what would have occurred? And how would our results differ if we

were to conduct this experiment again?

 Works Cited

Badurek, Theresa. Salt Tolerant Vegetable Gardening. UFIFAS Extension Pinellas County. Web. 15 Feb.
2017.
Hannah, James. "Farmers Use Radishes to Soften, Fertilize Fields." Phys.org. N.p., 18 Sept. 2009. Web. 22 Feb.

2017.

New Approach for Environmental Test on Livestock Drugs. UZH - Media, Universitt Zrich, 26 July 2016,

www.media.uzh.ch/en/Press-Releases/2016/New-approach-for-environmental-test-on-livestock-drugs-.h

tml. Accessed 5 Mar. 2017.

Qktheme. "World Losing Farm Soil Daily to Salt-Induced Degradation - UNU-INWEH."UNUINWEH. N.p.,

n.d. Web. 27 Feb. 2017. <http://inweh.unu.edu/world-losing-farm-soil-daily-salt-induced-degradation/>.

Stromberg, Joseph. What Happens to All the Salt We Dump On the Roads? Smithsonian.com. Smithsonian

Institution. Web. 09 Feb. 2017.

Thorpe, Gary S. Chapter 10 Impacts on the Environment and Human Health.Barron's AP Environmental

Science, Barron's, Hauppauge, NY, 2013, pp. 386386.

Zielinski, Sarah. Earths Soil is Getting Too Salty for Crops to Grow. Smithsonian.com. Smithsonian

Institution. Web. 09 Feb. 2017.

Rubric Categories 1 = Poor 2 = Improving 3 = Above Average 4 = Excellent

The report identifies The report identifies The report identifies The report situates
an overly broad or an adequately an adequately the students
Understand Context simplistic area of focused area of focused area of investigation of the
investigation and/or investigation in the investigation in the complexities of a
shows little evidence research and shows research and shows problem or issue in
of research. little variety in source some variety in research that draws
selection or poor source selection. upon a wide variety
source selection. of appropriate
sources.

Analyze Context A simplistic It makes some It makes clear the It makes clear the
connection or no reference to the significance to a significance to a
connections is made overall problem or larger context without larger context citing
of the overall issue. citing specific specific research
problem research

Understand and The report restates or The report restates The report Student is able to
Analyze Argument misstates information information gathered summarizes support their
(outside sources) from sources. It and information in places conclusion through
doesnt address and offers effective direct explanation
reasoning in the explanation of the or through
sources or it does so reasoning with the purposeful use of
in a very simplistic sources argument the reasoning using
way. (but does so statistical data as
inconsistently) evidentiary support.

The report identifies The report only The report in places The report
evidence from chosen addresses one of the offers some effective demonstrates
Evaluate Sources sources. It makes following: some explanation of chosen evaluation of
very simplistic, effective explanation sources and evidence credibility of the
illogical, or no of chosen sources or in terms of their sources and
reference to the evidence in terms of credibility and selection of relevant
credibility of sources their credibility and relevance to the evidence from the
and evidence, and relevance to the inquiry (but does so sources. Both can
their relevance to the inquiry inconsistently). be evidenced by
inquiry direct explanation
or through
purposeful use.

Evaluate Evidence Specific data, graphs, Specific data, graphs, Specific data, graphs, Specific data,
quantitative data cited quantitative data cited quantitative data cited graphs, quantitative
but no inferences but illogical or with weak inferences data cited and
made regarding the incorrect inferences to main theme. logical inferences
data and its made regarding the made regarding the
connection to main data and its data and its
theme. connection to main connection to main
theme. theme.

The report identifies The report identifies The report identifies The report discusses
Understand and few and perspectives multiple perspectives multiple perspectives a range of
Analyze Perspective that are vastly from sources from sources, making perspectives and
oversimplified from however, they are some general draws explicit and
sources. oversimplified connections among relevant connections
perspectives. those perspectives among those
perspectives

Experiment & Data Quantitative data Quantitative data Quantitative data Quantitative data
Collection poorly collected and collection attempted collected through collected through
poor or incorrect through experimentation with experimentation
connection to theme experimentation with a loose connection to with a strong
of paper. a vague connection to theme of paper. connection to theme
theme of paper. of paper.

Data Analysis and Quantitative data Quantitative data Quantitative data Quantitative data
Graphing analyzed using analyzed using analyzed using analyzed using
appropriate appropriate appropriate appropriate
calculations. calculations. calculations. calculations.
Appropriate graphical Appropriate graphical Appropriate Appropriate graphical
representation of data representation of data graphical representation of data
chosen to highlight chosen to highlight representation of chosen to highlight
significant details and significant details and data chosen to significant details and
support conclusions. support conclusions. highlight significant support conclusions.
details and support
conclusions.

Student demonstrates Student discusses the Student discusses the Student discusses
an obvious utilization of the utilization of the the utilization of the
Science as a process misunderstanding of appropriate scientific appropriate scientific appropriate
the utilization of the techniques and techniques and scientific techniques
appropriate scientific instruments. Makes instruments. Makes and instruments.
techniques and vague conclusions conclusions and Makes conclusions
instruments. Makes based on data evaluates their quality and evaluates their
incorrect conclusions collected. Proposes and validity based on quality and validity
that are not connected a simplistic statistical evidence. based on statistical
to collected data or suggestion for further Proposes further evidence. Proposes
statistical evidence. questions for study. questions for study. further questions
Does not address for study.
possible future
investigations

The report includes The report attributes The report attributes The report
In-text Citations many errors in or cites sources and accurately cites attributes and
attribution and in-text (in-text) used but not the sources used with accurately cites the
citation always accurately. few minor mistakes sources used.
(in-text) (in-text)

Works Cited Works Cited is The Works Cited is The Works Cited The Works Cited
incomplete inconsistent in style accurately references accurately
and/or format sources using a references sources
consistent style with a using a consistent
few minor mistakes style

The report contains The report is The report is The report

 many flaws in generally clear but generally clear but communicates
grammar that often contains some flaws contains some flaws clearly to the
Grammar interfere with in grammar that in grammar that audience (although
communication to the occasionally occasionally may not be free of
audience. interferes with interferes with errors in grammar
communication to the communication to the and style)
audience audience

(MLA Format) The written/technical The written/technical The written/technical The

style is not style is inconsistent style is mostly written/technical
appropriate for an and not always consistently style is consistently
academic audience appropriate for an appropriate for an appropriate for an
academic audience academic audience. academic audience

51 pts, 98% Mar 24-> Graded: 51 pts, 98%

52 pts Points Possible: