Kayla Cook

Myra Collins (Graphs Constructor) Alana Brown (Data Analyzer) Kayla Cook (Conclusion Coauthor) Alexis

Roberts (Conclusion Coauthor)

APES

Mrs. Kara Norris

2 March 2017

LD50 Radish Lab Report

Introduction

Chemicals are a part of everyday life in modern America, so knowing the levels of exposure to the

chemicals is vital. It is apparent that different chemicals have different toxicity levels, so the comparison of

toxicity levels is normally difficult. However, in order to compare the toxic potency or intensity of chemicals,

researchers have to measure the same effect. An example of a way to carry out a lethal dosage test is to measure

how much of a chemical is required to cause death. (CCOHS, 2017) LD50, lethal dose 50, is the amount of an

ingested substance that kills fifty percent of the the test population. LD50 is expressed in milligrams of a

substance per kilogram of body weight. (UC Regents, 2004). The LD50 is a way to measure the acute toxicity,

short-term poisoning potential, of a particular substance. Toxicologists, people who perform experiments on

lethal dosage levels, most commonly use mice and rats in the experiments. The LD50 test has continuously been

used in experiments for several different purposes. However, there is a large opposition to the LD50 Test. The

Pharmaceutical Manufacturers Association publicly opposed to the LD50 Test, after a study conducted was

shown to force 200 mice and rats an ingestion of toxic chemicals. A modified LD50 lab was suggested from the

U.S. Environmental Protection Agency to limit the number of rats to a maximum of 10 (NEAVS, 2011). Due to

LD50 killing large numbers of animals for experiments, it gained a lot of criticism, causing the UK Government

to ban the practice in 1991 (FRAME, 2014). While many LD50 experiments use animals, this particular lab tests

the LD50 for radishes. It would be useful to be aware of the lethal dosage of salt on radishes for agricultural
workers, and even for the population as a whole; agricultural workers would benefit knowing how much salt

concentration in the soil of radishes would produce the highest yield. Additionally, it is beneficial to the entire

population to understand the effects of excess salt concentrations in soils, and to be aware of the negative

impacts that over-salination of agricultural land (Zielinski, 2014).

Purpose

The purpose of the LD50 experiment is to decipher where half of the population will not germinate under

the increasing dosages of salt concentrations.

Hypothesis

If the salt concentration is raised for the seeds and the LD50 will be reached at 25% salt concentration

then the seeds will not germinate and the root length will deteriorate by a mean average of 1 centimeter as

compared to the seeds in distilled water, whose root length will grow by a mean average of 2 centimeters.

Materials and Methods

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.

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

2. 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

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

that sample

4. 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.

5. 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.

 6. 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.

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

8. For each treatment, calculate the mean radicle 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

Radicle Root Length (mm)

# seeds T=
Concentration germina Difference
Mean
of Salt ted (out in Radicle
1 2 3 4 5 6 7 8 9 10 Radicle
of 10) Length
Length
C-T
(mm)
Control
8 75 75 65 25 50 40 55 60 - - 55.625 0
0%

6.25% 8 55 55 25 37.5 70 45 57.5 15 - - 45 10.625

12.5% 6 50 45 30 40 20 25 - - - - 35 20.625
25% 4 15 5 10 5 - - - - - - 8.75 46.875
50% 0 - - - - - - - - - - 0 55.625
100% 0 - - - - - - - - - - 0 55.625
Table 1: Data of the experiment conducted on varying concentrations of salt solution to determine LD50
 Fig 2: Response of radish seeds to varying concentrations of salt solution

Fig 3: Response of radicle roots to varying concentrations of salt solution

% Salt Solution Mean Radicle (mm) Class Mean Radicle % Seed Ungerminated

0 55.625 68.18 20

6.25 45 61.7 20

12.5 35 67.21 40

25 8.75 44.04 60

50 0 3.79 100

100 0 0 100
Table 2: Individual and class data of the experiment conducted
 Fig 4: Response of individual and class average radicle roots to varying concentrations of salt solution

Fig 5: Threshold graph of ungerminated seeds at varying concentrations of salt solution

The LD50 lab that was conducted showed that the LD50 of radishes in a salt concentration is reached at

18.75% salt concentration for the individual data and the average LD50 for the class was 22.375. The control

group that had 0% salt concentration had the highest radicle root length at 75mm and had the most seeds

germinate at 8/10 seeds for the individual data. The group that was in 25% salt concentration had the smallest

radicle root length at 5mm and had the lowest number of seeds germinated at 4/10 seeds for the individual data.
The control group had the highest mean radicle length at 55.625mm meanwhile the 25% group had the lowest at

8.75mm for the individual data. The individual data correlated with the class data being that the highest amount

of radish seeds germinated was the control group or the 0% salt concentration group. In both the individual data

and the class data 100% of the radish seeds were unable to germinate in the 100% salt concentration petri dish.

The data concluded that when salt concentrations are above 50% nothing will germinate, and when there is no

salt present germination will be at its best.

Conclusion

In this LD50 lab, the experimenters predicted that the LD50 would be reached at 25% salt concentration,

but it was actually shown that the lethal dosage that killed 50% of the test population was actually at 18.75%

(between 12.5% and 25%) salt concentration. Additionally, the experimenters hypothesized that the root length

of radishes will deteriorate by a mean average of 1 centimeter as the salt concentration is increased. They

predicted the seeds in the control group would experience a root growth by a mean average of 2 centimeters. It

was shown, however, that the control group radishes had actually grown by an average of 5.6 centimeters. The

hypothesis, regarding the decrease in root length by a mean average of 1 centimeter as the salt concentration

increased, was very close to being accurate; the actual average of root length decrease was by 1.1 centimeters.

So this part of the hypothesis was only 0.1 centimeters off from the actual findings. Overall, however, the rest

of the hypothesis was proven incorrect. These results suggest that radishes grow best when there is no salt in the

environment. However, they are likely to germinate still, until they reach a salt concentration greater than 25%.

The results also suggest that other similar types of plants, such as yams and cassavas, are likely to experience

similar deterioration as the salt concentration is increased.

In comparison to the other LD50 labs, with regards to the germinations of radish seeds, it is evident that

the experiment conducted resulted in an LD50 of 18.75%, which is similar to the other LD50 levels. Comparing

the evidence between other groups, it seemed that the LD50 level commonly existed between 12.5% and 25% in

salt concentration, with the class average for germination levels existing at 20%. As the salt concentration levels
continued to rise, it is apparent that more and more seeds did not germinate and the root lengths of germinated

seeds continued to decline. Considering the individual data collected from the experiment is similar in LD50 salt

concentration levels, it is possible to conclude that a LD50 of 18.75% is primarily accurate because it falls in

between the 12.5% and 25% mark and is close to the class average of 20% salt concentration.

To measure the LD50 level of radish seeds that germinate, the levels of salt concentration differed.

Therefore, there were 10 seeds placed into containers with a salt concentration of 100%, 50%, 25%, 12.5%,

6.25% and 0%, which represented the constant. For materials, graduated cylinders were used to measure the

water and salt concentrations and equal layers of paper towels were used to soak up the water concentrations.

Ten radish seeds were then placed into each container of varying concentrations. One aspect of the experiment

that could have been improved to have more definite results would be to have smaller increments of change

between salt concentration levels. By making the increments between concentration levels, it would provide the

experimenter with a closer result as to where the true LD50 of the germination of radish seeds exists. As far as

causing error, the measurement of salt concentrations could have disrupted the results of the experiment. To get

each concentration level, the experimenters took the control level and the 100% concentration, to begin with,

and combined them to get the concentration of 50%. This process continued for each salt concentration level.

To get more accurate readings on the salt concentration levels, the experimenters could have found the actual

solutions of each concentration by adding salt to the constant rather than combining different concentration

levels of water. This would provide for a more accurate assessment of each level of salt concentration.

This particular LD50 experiment contains important results about the effects of excess salt in the

environment. Water enters a plants roots through the process of osmosis. However, if the level of salt is too

high in the soil, water will flow from the plant roots right back into the soil. This causes an issue, as the plants

are left dehydrated and it can lead to crop destruction (Queensland, 2013). Certain plants, including radishes,

are highly sensitive to soil with high salt concentrations (Shannon and Grieve, 1999). Radishes and other

primary producers that are sensitive to salinity are part of the larger food web in their environment. Many

different primary consumers depend on the radish as a source of food, and many different secondary and tertiary
consumers depend on the primary consumers as a food source as well (Ingham, 2000). So, salinity causes an

imbalance in the sustainability of an ecosystem and can lead to a decrease in the biodiversity of an area.
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