Question Section: Things you know that you dont know Question Section: Things you didnt know

ngs you didnt know that you dont know

How is Guinea pigs sense of smell?
How do we know that the maze can be used to determined olfactory
functions of guinea pig?
What should be the design of the maze?
What are some materials used in the experiment?
What are the experiments that scientists have completed in the past on
the sense of smell in mice?
How does the maze experiment give the information on Guinea pigs
sense of smell for foraging?
Can the Guinea Pig use their sense of smell for foraging?
How is the results of other scientists experiment on the mouse sniffing?

The experiment scientists have completed before

first journal;
Impaired sense of smell and altered olfactory system in RAG-1 immunodeficient mice
RAG-1-/- mice
Background information
The RAG-1-/- mice were prepared by by Prof. Hans Stauss, (University College London, UK) while control C57BL/6 mice. These two types of mice were
taken care in the same conditions. In the experiments, the scientists used male mice which have 7 weeks old and no 2 more tests per day. For the
embryos, they mated the mice in the evening, so in the morning the vaginal plug appeared means that it is successful mating. The embryos will be
considered 0.5 day old. All tests, which were approved by the local biological service unit at Queen Mary University of London, were performed in a
blinded fashion and according to the UK Animals (Scientific Procedures) Act, 1986.
Experiments
Buried food test
Purpose: to measure an animal's ability to smell volatile odors and its natural tendency to use olfactory cues for foraging.
Materials
RAG-1-/- mice experimental group
C57/BL6 mice control group
Environment, deepest and location of latent bait beneath the bedding, the type of the bait which is cookie control variable
Time that two types of mouse can reach the cookie dependent variable
Methods (testing protocol of 3 days)
First day, an odor familiarization exercise, the scientists use a clean mouse plastic cage (25 42 12 cm) containing 3 cm of fresh cage bedding to be the
place for 7-week male C57BL/6 and RAG-1 mice (6 mice per cage). Then they gave the 1 cookie for every 2 mice and left overnight. So they would
check if the cookies are gone or not for making sure that mice loved it.
Food deprivation on day 2 started one day before the test on 4 PM. The scientist removed food from the cage and let mice fasted overnight.
Testing on day 3 was started by moving the mice to the new cage with 3 cm deep bedding and let the mice make themselves at home in this new habitat
for 5 minutes for reducing the exploration the new environment. Next, the cookies was buried in the corner of the cage with constant length away from
every mice. The time was measured by the step of the mice which were counted in seconds. 15 minutes or 900 seconds was the maximum score (Yang and
Crawley, 2009).
Results
One of the main purpose of mouse for using the sense of smell is to forage, which means the buried food test is able to use for the experiment. Cookies,
the palatable food which is already tested from the palatability of the bait test that resulted no difference between RAG-1-/- and C57/BL6 because both
consumed the whole cookies. In the experiment, cookie was hidden under the bedding by measuring the length using one step of a mouse per one second.
As the experiment, the observation shows a significant five-fold increase in finding hidden bait. 203 s 77.7 in the RAG-1-/- mice compared to 42 s 18.9
in control group which is C57/BL6 mice.

Second Journal;
The Identification of Attractive Volatiles in Aged Male Mouse Urine
Experiment:
In this experiment, the scientists were testing about the aged males urine attraction toward the female. The attraction is greater in adult than in the
younger one.
Materials & Procedure:
- Animal:
The mice were bred at Hokkaido University and stay in 22C room which they will see light 12 hours and dark 12 hours.
- Urine Collection:
The urine will be collected from the mouse of the age 3-8 Months, 15-20 months, and 20 months. Each age urine will be collected from about 2-3
individuals. The urine will be store in the sterile tubes at -80 C.
- Ultrafiltration of mouse urine:
The urine were pretreated with Amicon Ultra-4 Ultracel-10k at 5C to remove volatile ligands and urinary proteins.
- Behavioral study:
They made the experiment to find out whether the age of male mice will made different attraction toward the female or not. By doing this, they used the
male mice urine to test. They collected the data by timing how much times will each female mice used to sniff different age of male urine. In this
experiment, they will used eighty-one virgin females mice with the age of 2 to 5 months to prove their hypothesis. About 3 to 4 mice were stayed in each
case. However, before the experiment, the mice need to practice for the 3 minutes habituation of sniffing the urine. The average time sniffing is 10.5
seconds. Therefore, in the real experiment, the mice with only 11 seconds sniffing will be recorded in the data. The experiment were conducted in the
rectangle cage and their were the two circular ports on each side of the wall. The ports were stuck above the floor for about 3cm high. Then, the glass
vials contained 0.5ml of urine of the different male age were put in each port. The urine that they used come from aged male, adult male, ultrafiltered
aged male, and ultrafiltered adult male.
- Result:
The average time for the mouse to sniff was 39.1 2.7 s for the both normal age urine and 36.8 2.2 s for both ultrafiltered age urine. In the
investigation for the normal urine, the female prefer the aged male urine more than the adult urine. In the ultrafiltered urine, there was no differences
among the female on both aged and adult male urine.

Point of including the literature review:

This information is important in doing our experiment because it help us to know more about the mouse behaviors. Also, from study other
experiments, the guinea pig running maze project can adapted other experiment to make it more reliable. We learned about environment factors that
might cause the mouse to frighten and will lead the the failure of experiment. Nevertheless, we learned to reduce the mouse stress and excitement during
the study. If we dont research about this experiments first, our experiment might get mess and collapse.
Cite:
According to research study of the Scents that Matter - From Olfactory Stimuli to Genes, Behaviors and Beyond by Rattazz, Cariboni, Poojara, Shoenfeld
and D'Acquisto and Identification of Attractive Volatiles in Aged Male Mouse Urine by Osada, Tashiro, Mori, and Izumi
Compare:
Before working on the experiment, both research have agreed on making the mice became habituation with the environment. Both of the study show that
they will place the mice in the experiment area to make sure that the mice can adapted to the new environment and will not frightened during the
experiment. From the result, in experiment 1, the average time that RAG-1-/- used are 203 s 77.7 and the average time for C57/BL6 are 42 s 18.9
which means that both of them can successfully find the food in the limited about of time(15 minutes). Also, in the experiment 2 result, the mice can used
their sense of smell to distinguish the different between the age of the male. The female will used their time to sniff the aged male urine much more than
the adult male urine. Therefore, from the result of both research, it can be concluded that the mouse sense of smell is important because it made the
mice to do many thing like foraging and surviving etc.
Contrast:
In the bury food experiment, the scientists test their sense of smell for foraging food in their bed and in the Urine experiment, the scientists test the
attraction of the male mice urine(different age) toward the female mice. After researching it show that sense of smell is one of the major ability for the
mice to survive.
Critique:
The literature review of experiment 2(The Identification of Attractive Volatiles in Aged Male Mouse Urine) is the most persuasive because the scientist
have test about the ability of odor distinguish between the mice(compare the attraction of the adult male urine, aged male urine, adult ultrafiltrate male
urine, and aged ultrafiltrate male urine). From the result of this experiment, it show that the mice can separate the different between odor.
Connect:
In the project, the goal is to investigate and analyse whether the experiment will support the research question of Can the Guinea Pig use their sense of
smell to help them foraging?. During the experiment, the result will show whether the guinea pig can find the food in the maze by using their sense of
smell or not. If they can find the food, it will prove that they can use their sense of smell for foraging. To connect with the research experiment(bury food
test), both of the experiment have a purpose to find about will the mice can used their sense of smell for surviving.
Cite: http://journal.frontiersin.org/article/10.3389/fnins.2015.00318/full
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2753229/
Compare: The experiment in both research is the same which is the bury food test.
Contrast: In experiment 1, the scientist didnt test for which food the mice will like the most. They only used cookies for the experiment. However, in
experiment 2, the scientists try several types of food which included cookies, cereal, chocolate chips, and food pellets.
Connect:
When the scientist put the mouse to the bedding cage for adapting, we think that they should give the mouse more time to stay in that cage(for about 2-3
hours). This is because if the mouse doesnt adapt to the environment enough, it will lead to an inaccurate experiment(the mouse might be frightened
while doing the experiment)
 Question Section: Things you know that you dont know
what question has not been answered yet about this topic?
how does your experiment help fill this gap ?
Question Section: Things you didnt know that you dont know
Can the Guinea Pig use their sense of smell for foraging?
what is the methodology used by other scientists?
what scientist use cite to help
What are some limitation of your design? How will data be collected?

Things you know and things you learned.

According to Royal Society for the Prevention of Cruelty to Animals science group (RSCPA), Guinea pig is the prey animals which is active in senses of new
sights, smells, and sound, so they have well-developed sense including sense of smell. Sense of smell is so important to a guinea pig, a social animals, to
interact with other guinea pigs and also humans.
The maze experiment helps to prove the ability of the sense of smell of guinea pig because, in the experiment, food is placed at the end of the maze in
the first trial to test how fast(in seconds) the guinea pig can reach the destination. Second trial, compared to the first trial, there is no food for this trial
to test if the sense of smell of guinea pig is effective by timing how faster or slower they will reach the end of the maze.
Methodology Literature Review
According to the buried food experiment, the scientists find the most tastable food for the mice. First step, they place the mice into the cages (6 mice
per cage) and then left the baits overnight which are cookies to test the palatability. Next, after testing the bait, the scientists put the mice to the new
cage to let the mice adapted with the environment. From the test, they suggest to put the mice in only 5 minutes but in our experiment the mice will
stay in the cage for 1-2 hours for adapting instead.
To create the maze our group have adapt the design from the psychologist man named Tolman. Adapted by using the curtain to blocked the way near the
exit. By doing this, it will test that if using the curtain to blocked the way to the food, can the mice pass through the curtain to find the food and exit
the maze.
Data Collections and Limitations
According to Yang and Crawley bury food test, they stated that the food deprived mice normally will find food within 1 minute and in their experiment,
they set the limited of times(12 minutes) for their deprived food mice. However, in the guinea pig running maze project, the guinea pig will not deprived
food. So, the time in measuring the guinea pig running maze will extended to 30 minutes each. In the experiment, one of the guinea pig will run the
maze that have no food at the exit and the other one will run the maze with the food at the exit. If the guinea pig cannot run the maze within 12
minutes, then that guinea pig result will record as failed.

This project faces many limitation. The guinea pig cant walk out from the maze, scaring of people and the maze is too small for the guinea pig. And the
way to collect the data is using watch to timing the time that guinea pig walk in the maze.

Sources: