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Group Member’s Names: Owen Berquist, Jordan Fritz, and Julia Gillespie
Date: 09/24/2018-10/02/2018
Period: 1

The Effects of Varying Salt Water Concentrations on Germination and Growth of Radish Seeds

Objectives

1. Calculate the effect of different amounts of radiation or salt concentration on the germination rates and
radical root growth of radish and lettuce seeds.
2. Determine the lethal dose of either radiation or salt concentration on radish or lettuce seeds.

Hypothesis

If the concentration of salt is higher, the less likely it is that the radish seeds will germinate successfully. Formatted: Indent: First line: 0.5", Line spacing:
Double
Seeds put in higher concentrations of salt will die, while those in lower concentrations will be more likely to

survive, however, those who do survive in the salt water will likely have inhibited radical root growth. If radish

seeds are placed in scenarios with different concentrations of salt, the lethal dose will be 25% salt solution.

Data and Analysis

Table 1

Radicle Root Length (mm)

Toxin Germination Mean Difference

Level rate Radicle in Radicle
1 2 3 4 5 6 7 8 9 10
Length Length
(T) (Control -T)

C 100% 5 11 8.5 4 2 1 12 3 4 9 5.95 0

6.25 % 80% 5 6 12 3 4 3 3 5 n/a n/a 5.125 0.825

12.5 % 50% 2 1 4 1 .5 n/a n/a n/a n/a n/a 1.7 4.25
25 % 0% n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a 5.95
 50 % 0% n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a 5.95
100 % 0% n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a 5.95

Graphs and Data:

Figure A
% Salt Solution Mean Radicle Class Mean % Seeds Class Average %
(mm) Radical (mm) Ungerminated of Seeds
Ungerminated

0 5.95 63.695 0 5

6.25 5.125 37.453 20 19

12.5 1.7 28.009 50 31

25 0 3.781 100 91

50 0 0.795 100 0

100 0 0 100 0

Figure B and C
Conclusion

After doing a dose-response study on radish seed germination in different concentrations of salt, it can

be concluded that the lethal dose of salt concentration for radish seeds is 12.5. These results proved to be

different than the hypothesis that the 25% salt concentration would create a mortality amount of 50%. However,

the experiment did accurately prove that the hypothesis that seeds in higher salt concentrations will die, while

those in lower concentrations will be more likely to survive, was accurate. Through the process of doing a

traditional experiment (control group vs experimental group), 10 seeds were put into plastic bags with paper

towels in them, which held different concentrations of salt water (100%, 50%, 25%, 12.5%, 6.25%, and 0%).

The seeds were then left in the salt water soaked paper towels for a week with no sunlight to help grow because
of the impenetrable drawer they were stored in. After a week, the germinated seeds’ radical roots were

measured in millimeters, and the number of seeds who did not germinate were counted. In the control (no use of

salt), all the seeds germinated, but as the salt concentration got higher and higher, fewer seeds germinated at

each concentration. At 12.5%, half the seeds did not germinate, representing an LD50 response (see Table 1). An

LD50 response/experiment is a test that figures out how much of a certain chemical does it take to kill half the

population (50%). Compared to the class data, the amount of seeds that germinated lies in the majority

compared to in the average radical root, the data collected is an outlier. However, while seeds were measured in

the correct units and were carefully removed from the paper towel, it is possible that some of the roots ripped

off when they were being removed, making different radical roots seem smaller than they actually were. Some

seeds also grew through the paper towel, so those radical roots may not have been measured because they were

not seen by the scientists. Another error that could have occurred is the seeds were close together in the paper

towel, causing them to intertwine with each other, making it more difficult to measure them. If the experiment

was conducted again, the seeds should be more spread out in the paper towel, and something should be between

the folds of the paper towels so the seeds to not grow through them. While the experiment futures the

knowledge about dose-response and LD50, it also raises questions about how ethically a study like the radish

germination study is on living animals, and how if there were multiple errors during a dose-response

experiment, how would that affect the human population as a whole. Overall, this experiment represents

something larger than radish seeds, and that is the effect certain levels of toxic chemicals can have on animals,

humans, and other important plants. Lethal dosage testing is extremely important in maintaining the safety of

society through prevention of fatal chemical consumption.

One article that was researched while doing this lab had striking similarities to this lab (Hernández, et

al.). The article discusses problems with the soil salinity of a landfill after contamination. A field was irrigated

with water from a landfill leachate. Soon, salt levels in the field and plants rose. Many of the other plants in the

landfill had died from too much salt, and the phytomass of the field was significantly altered. This shows the

harmful effects of excess salt in an ecosystem, and especially why you shouldn't irrigate a field with water from

a landfill. Keep in mind, this article was written in 1999, so the research on the harmful effects wasn't as
complete as it is today. Hopefully, this mistake wasn’t repeated since then. Overall, this article exhibits a real

world situation of the situation similar to the in-class lab. Both examples used higher and higher concentrations

of salt solutions, and in the end, the solutions had harmful effects on plant matter.

While LD50 experiments are done to find how changes in the environment can affect the organisms, such

as high salt concentrations in water affecting plants, they are also used to test chemicals that are for medical use.

Michael Gartlan and Henry Hoffman; who are both in the department of Otolaryngology at the University of

Iowa Hospitals and Clinics who specialize in head and neck surgery, deeming them credible; conducted a study

where they injected botulinum toxin type A (botox) into the throats of mice to find the lethal dose of botox that

can be done on humans for those who suffer from spasmodic dysphonia. Spasmodic dysphonia is a disorder

where the muscles in the larynx “spasm,” causing the voice to break and create a tight, strained voice that can

be difficult to understand (National Institutes of Health). People who have spasmodic dysphonia tend to have

voice breaks every few sentences also, making them even more difficult to understand, hence why they use

botox. Botox freezes the muscles and nerves in the place where it is injected, relaxing everything around it.

Most people use botox for cosmetic use, such as getting rid of wrinkles, but for people have spasmodic

dysphonia, it helps freeze the muscles in their throat to keep them from having voice spasms, making them able

to speak more clearly. However, for these people to be able to speak clearly all the time, they have to get botox

regularly, so people refreeze the Botox so they can use it at a later time. The FDA does not recommend

refreezing Botox at all due to the potency goes down, which is what Gartlan and Hoffman tested. They

concluded that the FDA was right. There was a 69.8% loss in potency after they froze the Botox and reused it

later on the rats, which can cause a problem for people who have Spasmodic dysphonia (Gartlan and Hoffman).

While they did an LD50 study like the radish seeds, they showed how a dose-response study helps people and

how it can help the human population live a quality life.

Not only do LD50 experiments test the effects of certain substances on plants, or chemicals ability to

affect humans, but they can also be used to test the effects of compounds used in the environment on the

animals that engage with it. Testing of chemicals effects on animals can then lead to further understanding of

how a substance will affect other organisms. A present study done by Jaya Raj, a professor at the India Institute
of Medical Sciences, and Mohineesh Chandra, who works in the Department of Forensic Medicine and

Toxicology at the India Institute of Medical Science, was created to determine the lethal dose of a combination

of “cypermethrin” and “endosulfan” chemicals found in commonly used pesticides on Wistar rats (Raj and

Chandra). Because both authors come from a highly reliable university, with access to accurate test data and

knowledge on the said topic of toxicity, the data and information derived from the experiment are very credible.

The chemical cypermethrin is a “neurotoxic synthetic pyrethroid”, primarily used as an insecticide, and

endosulfan is a “neurotoxic organo chlorine” that is also used primarily as an insecticide (Raj and Chandra).

When these two chemicals are mixed, they create a compound found in the pesticides and insecticides many use

for widespread agricultural purposes, and those that are largely found in the water, air, and soil because of

agricultural use. In order to conduct the experiment, 25 healthy adult male Wistar rats were put into an

experiment for 5 days, where they were housed in cages, with 3 or less rats in a cage, maintained under stable

laboratory conditions, given a standard dry rat pellet diet, tap water, and a 22 gauge-oral feeding needle holding

the chemical compound five times a day. The doses given to the rats increased each day, similarly to process of

increasing the amount of salt concentration in each bag of radish seeds for the in-class experiment, until there

was a 100% mortality. The concentration of cypermethrin and endosulfan consumed until 50% of the rat

population died, was 1320 mg/kg, meaning this was the LD50 dosage. Pesticides play an important role in

modern agriculture by being a way to prevent pests at low labor and low cost, however it is clear that they are

“potent pollutants” with “undesirable effects on non-target organisms” (Raj and Chandra). The toxicity of

pesticides on humans is estimated based on the test results on other animals used in lab experiments. This use of

pesticides, if increased too much and used in “additive” ways (such as cypermethrin and endosulfan used

together in the Wistar rat experiment), can cause deadly effects on humans and animals (Raj and Chandra).

Knowing the lethal dosage of a chemical substance is extremely important to understand before toxins are used

in the environment. Although a pesticide poisonous for rats is not necessarily as dangerous to humans, it is

important to have knowledge on the effects of chemical concentrations that are being used so widespread. This

experiment of pesticide consumption on Wistar rats is one of many LD50 experiments that measure the toxicity

of commonly used chemicals. It help shows how the testing of certain chemicals can affect organisms, similar
to the in-class experiment that tested the salt concentrations effects on the radish seeds. Ultimately, the use of

LD50 testing is an important way to ensure the safety of many organisms from lethal doses of chemicals.

There are many varying perspectives on how LD50 studies can be used to benefit different

species of organisms. LD50 studies can be used to discover medical and economical effects. Gartland and

Hoffman would say that a LD50 study is one of the best ways to find the lethal dose of medication for patients

who must take it everyday or at all. They would also argue that people should stay within what the FDA tells

them is not lethal and should be careful about refreezing medication. Hernández and their team would agree

with Gartlan and Hoffman about how LD50 is one of best ways to find the lethal dose of something, but they

would specifically argue how it can be applied to the environment. They would also remind companies that

collect garbage to be careful where they dump their waste water because if they are not, they could possible kill

off many plants in an ecosystem, getting rid of the biodiversity of an area. Haya Raj and Mondeneesh Chandra

would argue that the use of LD50 testing can save organisms in the environment, while still being agriculturally

beneficial. For example, the use of pesticides can be monitored to the point where they do not create fatal

effects for humans, animals, or plants, but also can create beneficial agricultural results. Overall, LD50

experimentation has been proven as a very reliable and useful way to prevent from fatal use of toxins in the

environment and as a way to create effective, healthy medicines, pesticides, and other commonly used items

with chemical substances found in everyday life.

 Works Cited

Gartlan, Michael G., and Henry T. Hoffman. “Crystalline Preparation of Botulinum Toxin Type a (Botox):

Degradation in Potency with Storage.” Otolaryngology-Head and Neck Surgery, vol. 108, no. 2, 1 Feb.

1993, pp. 135–140., doi:10.1177/019459989310800205. Accessed 02 Oct. 2018.

Hernández, A J, et al. “Soil Salivation from Landfill Leachates: Effects on the Macronutrient Content and Plant

Growth of Four Grassland Species.” NeuroImage, Academic Press, 23 Mar. 1999,

www.sciencedirect.com/science/article/pii/S0045653598003671. Accessed 02 Oct. 2018.

Miller, Lonnie. Salinization Lab, Teacher Lab Template. College Board, 2003. PDF. Accessed 02 Oct 2018.

“Spasmodic Dysphonia.” National Institute of Deafness and Other Communication Disorders, U.S. Department

of Health and Human Services, 15 June 2018, www.nidcd.nih.gov/health/spasmodic-dysphonia.

Accessed 02 Oct. 2018.

Raj, Jaya, et al. “Determination of Median Lethal Dose of Combination of Endosulfan and Cypermethrin in

Wistar Rat.” Current Neurology and Neuroscience Reports., U.S. National Library of Medicine, 2013,

www.ncbi.nlm.nih.gov/pmc/articles/PMC3702116/. Accessed 2 Oct. 2018.

Articles:
Owen Bergquist: https://www.sciencedirect.com/science/article/pii/S0045653598003671

Jordan Fritz: http://journals.sagepub.com/doi/abs/10.1177/019459989310800205

https://www.nidcd.nih.gov/health/spasmodic-dysphonia

Julia Gillespie: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3702116/

 Do not remove or mark below. This rubric is for grading purposes.

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0 = Missing 1 = Poor 2 = Above Average 3 = Excellent
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The report identifies an
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