Research Question:

Can the Guinea Pig use their sense of smell for foraging?

Introduction:

Do you ever notice that the mouse with small little nose has sharp sense of smell? The

reason for this keen sense is because of the olfactory receptor genes. The olfactory receptor is part

of the cell membrane in the olfactory receptor neurons. The role of it is to detect odorant which is

the compound that contain smell(Chapman & Bernays, 2011). The more it is, the better sense of

smell each individual will has. It is important to know because it is the natural process for all

animals to smell which it is helpful for them to find the ways back home and foraging. From the

scientific research, guinea pig was ranked on the top tenth animals that has the genes to distinguish

different smell. It has about 796 olfactory receptor genes. For the mice, it ranked on top sixth with

about 1,130 olfactory receptor genes. The mice have this much because they have variety of diet.

Moreover, the rat ranked on the second place with 1,207 olfactory receptor genes(Viegas, 2014).

This information is useful because it is important for the animals to forage by using their sense of

smell. Thus, in our opinion, for foraging, the food at the end of the maze will lead the guinea pig to

the exit.

The goal of this experiment is to find out whether the guinea pig can use their sense of smell

to lead them out of the maze or not. If the experiments result show that the guinea pig can find the

way out, then it will help to solve many problems. In Thailand, there are many rats and mice living

in the tunnel which this can cause problems. This is because flood always occur oftenly in cities.

When the flood come, human might get sick by the leptospirosis disease because the disease might

be mixed with the water. However, if the result be like our hope, the problem can be solved by trick

the mouse out of tunnel by their sense of smell. Another useful thing to use the mouses sense of

smell in the future is to use the mouse to detect disease. By doing this, the mouse genetic
engineering might occur to develop their new sense of smell. Therefore, from this experiment, it is

the guideline for other scientists to use this result in developing country and solve many problems.

Literature Review:

Impaired sense of smell and altered olfactory system in RAG-1 immunodeficient mice

RAG-1-/- mice (Experiment1)

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)

1. 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.

2. 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.

3. 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.

The Identification of Attractive Volatiles in Aged Male Mouse Urine (Experiment 2)

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.

Behavioural 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, ultra-filtered aged male,

and ultra-filtered adult male(Osada, Tashiro, Mori, & Izumi, 2008).

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 ultra-filtered age urine. In the investigation for the normal urine, the female

prefer the aged male urine more than the adult urine. In the ultra-filtered 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.

Knowledge Gab

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.

Overview of Experiment Details:

Edward Tolman is the psychologist. He also the first person who studies about rat by

investigate them by using the maze. He believed that individuals not only respond to stimuli, but

they also act on behalf of their attitudes, beliefs and will change conditions in order to achieve their

goals. He also have his partner name Honzik to joined the maze experiment. They help each other
build up the maze for that they can investigated the rat. According to the experiment, rats also tend

to process information rather than acting based on stimulus responses.

List of Materials:

Mouse:

Food Dry grass

Toys

Cage

Water bottle for animals

2 female Guinea Pigs

Stopwatch

Maze:

Feature board (optional)

Nails

Color (optional)

Fabric
Methodology:

For maze:

The experimenter will design and sketch the maze first. After finished designing the maze,

gather all the materials that will be used to make the maze. Next, measure and cut the feature board

into pieces and connect the pieces following the sketch that the experimenter designed by using the

hammer to strike the nails. Finally, use the fabric and sew it with thread to make a curtain and put it

near the exit of the maze.

For Guinea pig training:

The maze will be used in the experiment for two trials. In the first trial, the Guinea Pig will

run the maze that the food was placed at the end of the maze and there is a limited of time for it to

run the maze (about 20 minutes). In the second trial, another Guinea Pig will run the maze that the

food wasnt placed at the exit with the same amount of time(about 20 minutes). If the guinea pig

cannot run the maze within the limited amount of time, then it will considered as failed. Each trial

will be record to compare the result.

Maze Design and Justification:

This design have been selected to use in the experiment because the design is not too

complex for the guinea pig. In this experiment, the goal is to test the guinea pigs sense of smell. In

the maze, there is a curtain that block the way to the food and in during the experiment will find out

that can the guinea pig success to pass through the curtain to get through the food. So doing this

design will answer the question that does the guinea pig have sense of the nose good enough to

know that the food have been hide behind the curtain.

In the maze, the food will be placed at the end of the maze path to test the guinea pig sense

of smell and also being a reward for Guinea Pigs after it finishing running 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.

Ethical/ Humane Statement:

During the experiment, the guinea pig might get stress. So, to minimize their stress, the

experimenter need to pat them and let them take a free walk around the maze area. Moreover,

before the experiment, the experimenters need to put the guinea pig in the maze for adapting.

If the animal still afraid, then give them some snack and hold them closely.
 For the maze, after finished on creating it, let the maze dry by placing the maze in the sun

for about 2-3 days because it will have the smell of the glue which may cause the guinea pig to

have health problem. Moreover, let the maze dried so the guinea pig cannot eat the glue.

Timeline: of Experimental/Procedure:

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(15

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

20 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 20 minutes, then that guinea pig result will record as failed.

Proposal Conclusion:

This experiment will help with the understanding of the guinea pig sense of the smell and

knowing about their foraging behavior.

 References

Baggaley, K.(2016). 'Super Sniffer' Mice Engineered For Ultra-Powerful Sense of Smell. Retrieved

from http://www.popsci.com/super-sniffer-mice-have-ultra-powerful-sense-smell

Chapman, R. & Bernays, E. (n.d.). Olfactory receptor. Retrieved from https://global.britannica.com/

science/olfactory-receptor

Harvard University. (2014, August 3). Making sense of scents: Mice can identify specific odors

amid complex olfactory environments. ScienceDaily. Retrieved May 7, 2017 from

www.sciencedaily.com/releases/2014/08/140803193754.htm

McLeod, S. A. (2013). Tolman - Latent Learning. Retrieved from www.simplypsychology.org/

tolman.html

Osada et al., 2008. The Identification of Attractive Volatiles in Aged Male Mouse Urine. Chemical

Senses, 33(9), 815-823

Rattazzi et al., 2016. Impaired sense of smell and altered olfactory system in RAG-1

immunodeficient mice. Retrieved from http://journal.frontiersin.org/article/10.3389/fnins.

2015.00318/full

Viegas, J. (2014). 10 Best Sniffers in the Animal Kingdom. Retrieved from https://www.seeker.com/

10-best-sniffers-in-the-animal-kingdom-1768828254.html