Research Proposal

Introduction and Background Information:

Learning behaviour plays an important role of how animal behave and react in the

environment. When it comes to learning, the complexity of the brain will work by

experience, and this will make it possible to link it with a stimulus, which is exactly how

associative learning works in general. The functioning of the brain can be caused by animals'

diet and assumption throughout the day, more specifically, diets with high omega-3 fatty

acids. Omega-3 acts as a factor that is very essential for the brain to function, to grow, to

develop and of course, to increase the learning ability to its full potential (Ehrlich, 2015). The

way that animals can demonstrate how their learning gradually develop is through a series of

trial and error, or in other words, operant conditioning'. This simply combine the animals'

action together with an instantaneous reinforcement, where this reinforcement will slowly

punctured a behaviour into their memory and over a certain period of time, this behaviour

will turned to be their habitual occurrence, where the reinforcement is not in need. Operant

conditioning reflects on a wide range of behaviour, where each behaviour can be shaped to

desire, including attitudes, values and routines. In fact, with such reinforcement, it is a

method that allows the animals to complete daily practical significance (McLeod, 2007).

On the other hand, not only this experiment will analyse the animals' learning process

through operant conditioning, another learning behaviour in occurrence is what is known to

be classical conditioning. This basically involve action happen naturally from the animals

themselves without being taught initially, or it simply say, its their reflexive responses. The

way this works is 2 types of stimulus, conditioned and unconditioned, will be working

together. In this case, the food that will be provided for the hamster at each end of the maze

act as an unconditioned stimulus, as the hamster realised there is food, they will respond
without being taught initially. However, the maze is a neutral stimulus that the hamster might

take awhile to get use to it, as so the maze is a conditioned stimulus. When the food, an

unconditioned stimulus, combined with the eight-pathways maze, a conditioned stimulus, it

will urge a response of a running hamster. As a reflection, classical conditioning is an

essential concept of the animals learning behaviour because it emphasises the animals

interaction with their surroundings, naturally. This can then be adapted into ways that helps

prepare the animal from harm or any predictable biological events, including sexually

movement and interaction. As for that, it is also applicable in the treatments community of

therapy, such as systematic desensitisation (Applications of Classical Conditioning to Human

Behaviour, n.d.).

The importance of this will helps us determine how omega-3 impacted their brain,

reflecting their cognition through the way they behave and respond. On top of that, the result

from the experiment can be branched off into many other area of studying about learning

discovery, such as it can be applied to other species, including human.

Hamsters like to eat seed, grains, nut, fruits and vegetable. And naturally,hamsters

love to eat. The primary balanced diet for hamsters include Proteins used for growth and

tissue repair, Carbohydrates and Fats as an energy and warmth provider, Vitamins to keep

hamster in a good health, Minerals for building up strong bones, and water, the basis need for

body to function. Hamsters also need fresh and clean foods on a daily basis (Hamster Diet

and Nutrition, n.d). Not to mention, a pure and clean living space for both their mental and

physical health.

The major issue that is needed to be addressed is whether or not omega-3 can boost

the brain function and perform better cognition test. Omega 3 plays an important role for cell

membranes and is necessary for the helping to improve the brain function and behavior

(Julius, 2010). And, as omega 3 lead to a healthy life as mentioned above, hamsters are less
likely to get any brain diseases or behavior disorder such as on memory, cognition, and

aggressive behavior

Omega-3 helps reduce the damage caused by stress to the brain. This causes cognitive

problems and stress since omega-6 increases inflammation due to stress and cortisol prevents

formation of DHA a long-chain omega-3 fat. (Benefits of Omega 3 for Learning and

Memory, n.d). Omega 3 will help the hamster to release their stress from hormone called

cortisol. This cortisol is not good for hamster mental health.

Foods that are high in Omega 3 include egg, milk, oats. ( Hamster Diet and Nutrition,

n.d). Omega 3 contained food is very important because the hamster that is experimental

group must consume the omega 3. Omega 3 will help the hamster to do well on the

experiment than the hamster that consumes pellets.

The rat that consumes an enriched (more) omega 3 contained food gain more weight

during the time of a restraint stress; on the other hand, rats that consume a diet of normal or

lacked omega 3 tend to lose weight over the same period of time under the same condition.

Notice that all the rat had the same body weight at the start of the experiment. The liquid

analyzes showed the amount of DHA concentration is proportional to the omega 3

consumption of the diet and as the chronic restraint stress procedure reduced the

incorporation of DHA into brain membrane phospholipids, it means that the more omega 3

consumed, the better its to deal with the stress in order to maintain a suitable DHA

concentration. (Influence of Omega-3 Fatty Acid Status on the Way Rats Adapt to Chronic

Restraint Stress, 2012). Omega 3 rat will gain weight in stressful conditions. But rats that do

not get omega 3 will have lower weight conditions because of consumption of pellets. Omega

3 is found in foods that cause weight gain. As the result came out, too consumed the omega 3

at a good level will cool down the stress that causes the change in DHA concentration.
 From the above, omega-3 plays such a big impact in the brain processing system, so it

is crucial to know whether or not providing the hamster with foods high in omega-3 will

results in better performances over an assumption of a normal diet. As so, the research

question is Which hamster will be able to run through a maze faster, with one hamster fed

foods high in omega-3 and other other a normal hamster diet?"

Hypothesis:

If one hamster is fed with omega-3 based diet completely and the other hamster is fed

with normal hamster diet, then the hamster fed with omega-3 based diet will be smarter and

have better memory to walk through the maze because omega-3 has been known to improve

many organisms mental ability.

Literature Review :

1. Based on Acta Physiologica Hungarica, the experiment The effect of omega-3 on

cognition in hypothyroid adult male rats stated that brain function involved the help from

Omega 3 and thyroid hormones. In this research, the experiment focused on the effect of

omega three on cognition in hypothyroid adult male rats. Omega 3 has played a major role of

brains function and also be a component of neurone's membrane.

Materials and methods:

Animal: Adult male albino was used for the experiment.

Experiment: 30 rats were divided into three groups: 1) Control group will receive water at

first and stop giving them at last,2) Hypothyroid group will receive a carbimazole which is an

antithyroid drug for six weeks, 3) Omega-3 treated group will receive both carbimazole and

omega three from week 2-6. All three groups will get food and a supplement equally.
The Control Variable of the experiment is the food, water and environmental condition,

meaning these are factors that all rats in the experiment will encounter. Behavioural testing

was then conducted to see the behaviour of rats using Eight arm radial mazes. In this,

initially, plenty of food was placed all over the maze so that rats will get familiar with the

maze. Later on, reduce the number of food in the maze and allow some arms to be free from

food and some that contain the food. Rats will be expected to remember the place of the food

placed in the maze as a measure of their memorising ability. Entry into arms that do not

contain the food will be counted as working memory errors. A re-entry to any arms that the

rat has eaten the food from will also be counted as a working memory errors. Likewise, this

observation was made each day for four consecutive days for analysis part. Another aspect of

testing the working memory of rats was Passive avoidance test, a test that consists of Dark

and Light compartment; Dark compartment would cause an electric shock on the rat. In this,

rats were allowed to explore the compartment and would receive an electric shock whenever

they enter the dark compartment. Rats were expected to enter the dark compartment less than

before. A re-entry into the dark compartment after being electrocuted is counted as a working

memory error. The more times the rat enters, the larger the magnitude of the working

memory errors. In latency for entering the dark compartment was recorded. Behavioural

testing: Experiment shows a decline in working memory errors in all three groups of rats.

However, The least number of errors were noticed in the control group whereas the most

were observed in the hypothyroid group. Passive avoidance testing: Experiment shows that

the control group had the fastest reaction time as compared to all other groups. The

hypothyroid group had impaired avoidance learning and showed the very slow reaction when

entering the dark compartment. The omega-3 treated group, however, showed an immediate

reaction time. The experiment conductor agrees and believes, from the result of the

experiment, that the rat was not receiving any additional drugs or food perform more
effectively in maze running, and other memory required tasks than the rat thats given

additional drug or food. This is because the experiment results show that control group made

the least number of errors when put to perform, a particular task. Additionally, the author also

believed that the most effective way to conduct the experiment is by using the eight armed

maze. This is because eight armed maze allows the conductor to test the memory of, the rat

more effectively. The author the would disagree that any additional change in food or any

consumption would help increase the performance of hamster running in the maze.

This experiment requires the proficiency in petting the animals and lot of experience

in using laboratory equipment. A small mistake in manipulating any of the variables in the

experiment would result in a potential error and inaccurate measurement results. The ability

to make the hamster feel comfortable in order to produce accurate results is one of the

requirements for conducting the experiment. Therefore, the downside of the experiment is

needed for skills in working with animals. The pros of the experiment, however, is the

method of finding the accurate results. By allowing the rats to walk freely in a maze, it can be

observed clearly the rat that would be able to find their food more quickly with making

errors.

The group experimental design is very similar to the experiment done in the literature

review. Experiment to be performed will be conducted in an eight-arm maze, in which the

hamsters will be the determiner of the experiment. One similar factor between the experiment

done and the one to be performed is the placement of food on each arm to determine the

working memory of the organism. The re-entry into the arm is counted as an error. However,

the different variable between the two experiment is the organisms. Another aspect, in the

planned experiment, a slight bent edge was added to help ensure a more accurate outcome.
The hamsters will rely upon will not use their eyesight to see the food on each arm, but rather

on their memory to work remember the location of the food.

2. Omega-3 is an essential nutrient in daily life. According to National Centre for

Biotechnology Information under the headline Influence of Omega-3 Fatty Acid Status on

the Way Rats Adapt to Chronic Restraint Stress state that Omega 3 is important for the

neurone function, cognition and also help to prevent the hamster from stress or moody.

In the experiment, the researcher creates a repeated restraint stress to the three groups of adult

rats; the first group is the control which is the group that will get high-oleic sunflower and

rapeseed oils. The second group is the omega three deficient group whose will get only

sunflower oil. And the third group is an omega three enriched which get sunflower and tuna

oils. The reason that the researcher has two experimental groups is to compare the results

from the two group and the control group. If the experimenters had only gotten data from the

control group and enrich-omega 3 group, then the result will only allow demonstrating

whether omega 3 is better than normal diet. However, if the omega three deficient group is

added, then the result that is gotten will help determine the cost and benefit of omega 3. And

also help the reader to do further research about the amount of omega 3 in the food and the

effect on animals behaviour and brain function.

The control variables include room temperature which must be at +/- 22c, the life cycle is

12-12 where the light is 7am-7pm, and water is given as desire. The experimenter has to

make sure that all rats were keep under the same condition to prevent from bias result.

For the experimental method, its started by giving each of Wistar rats groups with their

assigned food with an ad lib water. These females will then force to mate after theyre fed
with this routine for two weeks. By giving the assigned food prior two weeks before the

female rats were mated, their offspring will be able to absorb those nutrients from mum since

theyre embryo.

After the female gives birth to the offspring, the male offspring will be distributed randomly

into three groups. Each group will be assigned to eat one of the three type of foods (control,

omega-3 deficient, and omega-3 enriched.) Then they will be sent to live in the same cage

with the mother who has the same diet. In the first three months, the rat will get 25 g of food

each; then theyll get 20 g of food a day until their age is six months. The weight will be

measured every two weeks and will be weight every week during the stress procedure. The

weight is the measure to make sure that all the rats were safe and alright. Since the rats get

the same type and equal amount of food, their weight supposes to be the same or not much

difference. When its time for the maze running an experiment, the weight helps to make sure

that they're raised under the same living condition.

Body weight result comes out that all rat has a weight between 6.5 +/-2 at birth. After six

months of particular dietary, the omega-3 enriched group gain 10% more weight than the

other two groups. Even though the rats in all group has lost the weight during stress period,

the omega-3 enriched group gain weight faster than the other two groups.

When the rat reaches the age of six months, it is time to enter the wire mesh restrainer. Each

rat will be placed in the restrainer for 6 hours a day for the 21 days to measure their stress,

and then they were grouping the stress and unstress rat into six categories include stressed

control group, unstressed control group, stressed omega-3 deficient, unstressed omega-3

deficient, stressed omega-3 enriched, unstressed omega-3 enriched. By separating the rat into
stress and unstress group, the experimenter can learn the connection between the diet of rat,

the stress after rat going through the stress procedure, and the ability to run a maze.

Finally, the rats were grouped into six categories; each rat was placed into six arm maze to

test their behaviour. It reveals that in omega three deficiency groups, stressed rats had a lower

motor activity score significant than unstressed rats. However, in omega three enriched group

has no difference in score between stress and unstress group. Moreover, the stress omega

three enrich group have the higher score than other two stress group. This result of the rats

weight shows that the omega-3 enrich groups can recuperate faster than the control and

deficient omega 3 group. Moreover, the motor score of behaviour also supports that omega

three enrich have a positive effect on rats behaviour.

This experiment requires a lot of time to do the experiment since it is testing on the

first generation of male rats that born from a female that has to eat an assigned food for two

weeks before mating. And those male offspring will be used in the experiment for at least six

months. To test the behaviour of the rats, it requires a motor which is the object that couldnt

find in the daily life. The good point about this experiment is that there were the different

experimental group which is omega three enrich and omega three deficient which help to

determine the effect of having and not having omega 3.

From studying the two experiments, both experiments feed different food but have the

same control group rats. The experiment is focused on the use of omega-3 as an experimental

group. But the first one used hypothyroid as another experimental group. Goals of both

experiments were to determine the effects of different nutrient from different foods effects

on the rat's body functioning. The result of the first experiment shows that each group

perform better than the others in different situations given . However, on the second
experiment it shows that the control group spent the lease time to get out of an open arm

maze. The weight of the rat of this group also reduce which mean they are more stressed.

After discussion with the group member, a conclusion has been made which is to use normal

diet for a control group and high omega-3 food as a experimental group. By setting it this

way, the result were able to compare how omega 3 really effect the hamsters cognition level.

Then a test will be made to examine the effect of diet through the ability to memorize the

arms of the maze.

Knowledge Gap:

In the literature review, information regarding rat has been found. In the first

experiments found, rats are categorised into groups and given different environment

conditions to thrive: Hypothyroid group, omega-3 treated group and control group. From this,

it was to be determined the rat that would perform best in the maze and the passive avoidance

test. Based on the results, it has been found that control group perform best in both

experiments. In the second experiment, the result shows that rat that consumes omega three

food able to gain weight faster than the deficient omega 3 and control group. And these

groups also did best in running a six arm maze.

In future, our experiment, however, focuses on finding how hamsters would be

affected by different food condition: Omega-3 rich and a normal diet. The method for

determining the better hamster would be using an eight armed maze. Therefore, the

knowledge gap between the future and experiment and the experiment done in the past is the

differences in organisms.
Research Question:

Which hamsters will be able to run through a maze faster, with one hamster fed a raw

vegetables and the other a normal hamster diet?

Overview of Experiment Details:

In this project, the experiment will be based on two hamsters, in which one of them

will be fed with a high omega 3 food, but the other one with pellets. At that time the

participants will then make a maze for the hamster out of balsa wood. The maze is required to

be taller than the hamster; otherwise, when participants let hamster run through the maze it

may cheat by climb up the wall (balsa wood). At the finish line participants will put some of

the hamster foods in the food dishes in order to attach the hamster . First, one of the hamster

will be chosen randomly. So if participants choose the first one that fed by high omega 3 to

run the maze first. Participants will timing the time it takes to complete the maze and record it

down on the data table. Then, the same method will be repeated for the hamster that is fed by

pellets and write the result down. Participants will repeat the experiment for about 3-5 times

before finding the average for each of the hamsters to see which hamster can run through the

maze faster.

List of Materials:

1. 2 Male Hamsters

2. Control groups food : Oxbow food, hard boiled egg, milk

3. Experimental groups food : egg, milk, oats

4. Hamster facilities: Hamster cage, bedding, food dishes, water bottle, toys

5. Maze : balsa wood, scissor, tape, glue, stopwatch, ruler meter, saw
Methodology:

1. First of all, put the hamster in their own cage and let them run and play around the

cage so they can feel more comfortable with the new place.

2. Then, it is essential to building a good relationship with the hamsters. Placed the

hamsters food on the hand before putting the hand into the cage and let hamster run

to eat the food from hand. By doing this, the hamster will learn that anytime the hand

was placed in the cage, and it will be their feeding time.

3. When the hamster starts to be familiar with the hand, it is time to pick it up from the

cage and put them in the maze. Before picking it straight up, making sure that it dont

frighten. By doing so, just nudge it gently with a finger before pick it up.

4. Feed the two hamsters with their assigned food as much as they desire for 3days. The

control hamster will be fed with the hamster pellets. And the experiment hamster will

be fed with food that is high in Omega 3 including egg, milk, oats. The food will be

placed at each end of the eight arms. They will have to run to the end of the arm in

order to eat the food. However, each end will be placed more than one piece of food.

5. For the rest of the experiment, well give only 85% of its original consumption (for

both hamsters)

6. On day 4, put one piece of food on each end of the eight arms. Place hamster in the

centre. And use a stopwatch to the timing for 10 minutes. (Repeat this step with

another hamster)

7. On day 5, do the same method as day four but start recording the time and the number

of arms that the hamster goes into. Plus, record the error that caused by hamster going

into the arm that has already been to. Repeat this for day (6-8)
Maze Design and Justification:

Eight arm maze with slight bend at the end of each arm: The reason for choosing this

maze design is to test the hamsters memorizing ability of whether or not they can

remember the path they went and not going in the same path again. Not only is this

design effective in testing hamsters memory in a different way but it is also unique in

terms of their shape.

This maze has center platform with eight arms protruded radially outward with a

slight bending at the end of each arms. The purpose of having multi arms is to test

whether or not hamsters recognize the path the have been to and not going there twice

or more. The more intelligent hamster would potentially be better at recognizing the

paths and thus less likely to be tricked. A slight bending of the end of each arms is for

hiding the food from hamsters. So hamsters will not decide on taking a certain path
 based on what they see but rather rely on their memory to decide which path they

took to not repeat going there.

Methodology Literature Review:

Based on Research Animal Resources stated that mice could be moved from one place

to another with the bare hand, it has to be gently and slow because mice are quilted

animal. People might get bitten or make them afraid easily. A rat can be held by

thumb and finger. Its position can either hang quietly or turn around to sit on the palm.

Therefore, to carry hamster from the cage to the maze, the hamster must feel calm and

not scared. (Cooper, n.d.)

From the research, before hamster is pick up or touch, it should be familiar and aware

of the presence of another thing first (Shukla, 2011). By doing this, the hamster

should be known that when their body contact with the finger, it means no harm. And

that is when people were going to carry them. With the frightening condition, the

hamster can be stressful. To adapt this to the experiment, the hamster should learn to

inure with the finger and hand. Moreover, from this research, it holds that hamster

will be familiar with one another, especially the same sex. However, the hamster will

be very aggressive toward strange animals. To conclude, hamster shouldnt be in the

same area or same cage with the difference sex, species.

According to the Journal of Experimental Psychology: Animal Behavior Processes,

rats initially tested on eight armed mazes performed efficiently in remembering the

samples placed at different arms, attempting not to visit the arms they have been

through. (Mcdonald, 2014)

 Procedure: The first stage is called exploration The rats were given an unlimited

food for the whole nine days. (15mins each day). No food represented in apparatus in

this day.

The second stage is the Free Choice where rats were given a specific amount of

food in order to bring the weight down to 85% of ad lib food (* ad lib food mean the

food as much as desire) This daily were given to rats for the whole experiment. For

the day 10-19. Purina Lab Cow ( the type of food) were placed at the end of each arm.

Then each rat was placed at the centre of the maze and was given 10mins or 16

choices.

The next five days, the food was placed on one of the eight arms of the maze. If the

rat was influenced by the presence of food on that particular arm, then the rat should

return to the arm after fewer choices than that when food is absent.

For the last three days, the odour of either lotion or old spices in the centre and all the

arms. (The author hypothesised that If the sense of food is still influence the rats to

run through the arm, then it will be less accuracy than the free choice.

Results: For the first few days, the rats only run around the centre. Over the next few

days, the rat started slowly venturing around the arms of the maze; some are hesitant

and pause at the edge of the platform. However, once stepped on one of the arms, they

went to the end before returning. It was noticed that wider arms are preferred than the

smaller arms are.

For the free choice result, there has been an increasing in some consuming. For the

first few trial, the rats weren't eating the platelet even though they can smell and see it.

After a few days, all the better were eaten. As before, the rats will stop at the edge of

the arm before choosing it, and once they've chosen, theyll run until the end.
 For Rebait, replacing food on one of the arms had no apparent effect on the accuracy

of choice behaviour. The rats would at least visit eight arms of the maze before

choosing the arms with the food at the end of it anyway.

For added odour, in the beginning, the rats were reached to the scene of lotion, and

some of them were walking around the centre while some weren't eating the pellet at

the end. In the second day of odour added, all the rats were run to the end of an arm

and eat all of the pallets. In conclusion, the odor has a little effect in an accuracy of

the choice behaviour.

For all procedures, in general, rats tended not to choose the same arms chosen,

although there were very few cases in which two to three adjacent arms were elected

in order. The rats often responded in a clockwise or counter-clockwise and 90 degree

to the arms just chosen.

Inspiration: From reading the experiment about the rats ability to run a maze in

different conditions, a designs ability to test the rats performance is in a unique way.

It tested the effects of different factors that could potentially change the way rats walk

through a maze. Therefore, the experiment would like to adapt from this experiment

in these ways to find out about the rats memory and ability to adapt to a uniquely

designed maze.

Ethical/ Humane Considerations:

In order to begin an experiment, it is very essential that the participant considers

safety as the number one priority, this is due to the fact that it is such a sensitive subject. In

fact, for this particular experiment, the participant have to be extra careful because they will

be working with animals. After many brainstorming and discussions, a decision has been

made that the maze body will be made out of Balsa Wood because it is sturdy, lightweight
and most importantly it has a soft touch to it. This soft texture will ensure that the wood

doesnt have any sharp edges or small bits of splinters exposed. In fact, to better guarantee

this point, the wood should be passed through a filing process using sandpaper to get rid of

any wood splinters that may have to stick out, which may harm the hamsters sensitive skin.

Moreover, for the food, it is sure to be clean with no toxic or dangerous chemicals in contact.

Every raw vegetable used in the experiment will be fully washed with care before providing

it to the hamster. As for any other materials, participants must make sure it is safe by cleaning

it before use. While the materials are under consideration, participants should ensure that the

hamster itself are not stressed during training. To do so, participants should go back and forth

to their cage and play with them, so they are not bored and stressed out. Also, playing with

them routinely will allow this hamster to be familiar with human behaviour so they are not

panicking on the actual experiment day. On the base of playing with them, some precautions

must always be listed. To minimise the hamsters stress as much as possible, before picking

them up, participants should nudge it gently with a finger and gradually pick them up,

handling them carefully with both hands. This part must not be overlooked because if the

hamster are distressed or frightened anyhow, not only it will be difficult to perform the

experiment, but it can also lead the participant to a painful harm. Another point that every

participant should keep in mind is to perform the experiment quietly so that it wont disturb

the hamster, mentally. As well as during the experiment day, any toys or form of

entertainment must be separated due to the fact that the hamsters can get overly excited and

jump around causing more stresses. Lastly, of course, for participants own safety, it is

important that the participants always keep clean, both before and after, the experiment by

washing the hands thoroughly. This will ensure that no toxic chemicals are intact from the

experimenter and pass on to the animals. All performance must be under adult's supervision

so any immediate problems occurred can easily be handle. If for some reason any of the
group members got bite by the hamster during the experiment, they must keep calm, so it

doesnt cause any injuries or anxiety to the animals while calling for a supervisor as soon as

possible.

Data Collections and Limitations:

Use the learning record form whenever anything is making a progress with the date

stated to keep track of your learning overtime.

Day Data Hamster 1 Hamster 2

1 Number of errors

Time

Record the number of times each hamster visits each arm of the eight armed maze. A

visit to the same arm twice or more is counted as an error. Notice a number of times of error

done by each hamster and compare. The hamsters that made more mistakes will be

considered as less intelligent. The intelligence level of each hamster will determine which

independent variable (food) of the experiment is the most effective for them as each hamster

maintains different diet; the one was eating raw veg or the other with the normal diet. If the

number of errors is the same for either hamster, then measure the amount of time taken for

each hamster to complete all the arms.

Limitation:
 The smell of the food can guide the hamsters way. During the experiment, hamsters

might be misguided by the smells of the food, which weakens the experimental design

because it bases on hamsters using their memory to lead their way.

Conclusion:

This experiment will help us to understand the relationship between different types of

Hamsters food-Normal hamsters diet and high omega three food- and the Hamsters

learning ability. From observing the hamsters behaviour in the maze, the ideal food for

hamsters could be identified. Knowing the type of good quality food for hamsters in the real

world will help in the development of welfare of hamsters.

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