Madison Gilbert (Technical Manager) & Megan Martin (Project Manager)

Collaborators: Vincenzo McEvoy (Safety Manager) & Emma McManus (Materials Manager)

AP Environmental Science

Mrs. Norris

12 February 2016

LD50 Lab Report

Introductory

A bioassay is a biological organism used to test the toxicity of a chemical and is commonly used in

environmental studies and by drug companies testing new products. In this lab radish seeds act as our bioassay

and test the toxicity of our sodium solution and its effect on plant growth. To estimate the acute toxicity of

chemicals on a bioassay, toxicologists use LD50 values and these LD50 values are calculated from

dose-mortality curves. When we have a chemical, for example, silver nitrate, (Oral- Rat LD50 = 50 mg/kg) this

means that an oral dose of 50 mg of silver nitrate per kg of body weight will kill 50% of the test sample of the

bioassay. Another chemical such as sodium iodine (Oral- Rat LD50 = 4340 mg/kg) shows that an oral dose of

4340 mg of sodium iodine per kg of body weight will kill 50% of the test sample of the bioassay. This also

means that compared to the silver nitrate, sodium iodine is a far more dangerous chemical, killing greater

amounts of the test sample (Hoyle, Brian). To successfully determine how seeds respond to various

concentrations of salt in this lab, a dose- response curve is displayed to plot the relationship between the salt

concentration and its effect.

Problem / Purpose

The objective of the LD50 lab is to determine if different concentrations of salt solution affect the

germination and radicle root growth of radish seeds.

Hypothesis
 A solution with a high ​concentration of salt​, such as 24 g/L or 16 g/L, will increase the ​germination

and radicle root growth​ of radish seeds, as compared to regular ​distilled water​, which will not have a drastic

effect on the germination or root growth of the radish seeds.

Data and Analysis ​-2 dose-response graph?

-2 incorrect analysis

T=
Concentration of % Seed Mean Difference
Salt Germinated 1 2 3 4 5 6 7 8 9 10 Radicle in Radicle
Length Length C-T
(mm)

Control 100% 45 48 20 28 40 25 24 23 18 35 30.6 0

0g = 0%

1.5g = 6.25% 100% 27 27 10 34 35 25 11 33 26 38 26.6 4

3g = 12.5% 100% 35 35 33 28 29 21 10 13 22 17 24.3 6.3

6g = 25% 20% 4 1 0 0 0 0 0 0 0 0 .5 30.1

12g = 50% 0% 0 0 0 0 0 0 0 0 0 0 0 30.6

24g = 100% 0% 0 0 0 0 0 0 0 0 0 0 0 30.6

Table 1​: Radicle Root Length (mm) of Each Concentration

​Fig 1​: Group and Class Radicle Length Results

This is a collection of observations, measurements, multiple trials, a data table, and a line graph showing

that the more salt in a solution, the less likely for the radish seed to germinate and grow. Observed during the

experiment was the display of vast growth among concentrations consisting of little or no salt, thus proving

against the group hypothesis (a solution with a high concentration of salt, such as 24 g/L or 16 g/L, will increase

the germination and radicle root growth of radish seeds, as compared to regular distilled water, which will not

have a drastic effect on the germination or root growth of the radish seeds). The reason higher salt

concentrations did not allow radicle root growth was because early seedling growth is most reliant on

freshwater, allowing the salt-tolerant seeds to thrive most and grow. This is presented by the observation: in

concentrations of 0%, 6.25%, and 12.5% of salt, the percent of seed germination was 100%. As for

concentrations of much higher salt, 50% and 100%, 0% of seed germination resulted. From this information,

more specifically the table, we can conclude that the LD50 of salt for these seeds is a dose of approximately 12

grams of salt in the concentration. The mean length of the radical for each of the germinating radish seeds in the

order of no salt to 25% salt is 30.6 mm, 26.6 mm, and 24.3 mm (calculated by adding all radicle root lengths for

each concentration and dividing by 10), not including the salt concentrations of 50% and 100% which resulted

in 0% mean radicle length. When comparing the group’s mean lengths to the whole class, as visually shown in

the graph, it can be concluded that results are not far off, meaning that all groups that conducted the experiment

generally came to the same consensus. Potential causes of error in this experiment could be caused by errors in

calculation, for example measuring radicle root length incorrectly. The measuring of the sprout shoot and

cotyledons, rather than only measuring the root is a contributing factor to experimental error.

Conclusion:

The results from the radish seed lab show that radish seeds experience the most germination and longest

radicle root growth when exposed to water, the control. As more and more of the salt concentration was added,

less and less of the radish seeds germinated, and their radicle root growth continued to decrease, as compared to

the control. The radish seeds experienced an LD50 when six grams of salt concentration was added to the petri
dish, indicating that fifty percent or more of the seeds were dead or ungerminated. Overall, the salt had a

negative impact on both the germination and radicle root growth, decreasing them both. This data proves the

hypothesis wrong. Instead of the salt improving the germination and radicle root growth of radish seeds, it had

the opposite effect. This effect can also be seen in many types of crops due to the stress an excess amount of

salt causes plants (Melbourne University). ​¶ ​This type of experimentation could be useful in any other scenario

involving the growth of a plant and the addition of salt; this could maybe be reflected by the growth of sea

plants and acidification, or of plants growing by an agricultural field, that have experienced pesticide run-off.

Incorrect measurements may have been the source of some error. This can be solved by having only one person

complete the measurements in the future. Another source of error could have been, people accidentally

switching the petri dishes from one class to another; although this is very unlikely due to the labels on all petri

dishes and containers. The setup of this experiment could have been more effective by assuring that all radish

seeds were placed far enough away from each other as to prevent their roots from growing together (as this did

occur with a few radish roots in the experiment). Contamination could have been prevented more effectively by

taping the petri dishes shut. The equipment used in this lab was as precise as possible. The purpose of this lab

was achieved because the data shows that the addition of salt concentration does affect the germination and

radicle root growth of radish seeds; salt affects them negatively, but it does affect them, so the purpose was

achieved. The results of this lab were fairly similar to the rest of the class, so the conclusion that this lab is

accurate can be reached.

 Works Cited

University of Melbourne. "Plants Also Suffer From Stress." ​Science Daily​. Science Daily, 2015.

Web. 16 Feb. 2016.

Hoyle, Brian. "Bioassay." ​- Test, S, Compound, and Organism​. Net Industries, 2015. Web. 15

Feb. 2016.