COGNITIVE ABILITY ASSESSMENT OF ALBINO RATS (RATTUS ALBUS)

BEFORE AND AFTER EXPOSING IT TO VAPORIZED ELECTRONIC CIGARETTE

JUICE (NICOTINE-FREE)

An Investigatory Project Presented to the faculty of Calasiao Comprehensive National

Highschool

In Partial Fulfillment of the Requirements of the Special Science Program

Markkjil C. Tayaba

Proponent
 TABLE OF CONTENTS

Title Page………………………………………………………...………………….……..i

Abstract……………………………………………………………………………………ii

Acknowledgement………………………………………………………………………..iii

Chapter 1: THE PROBLEM: RATIONALE AND ITS BACKGROUND

Background of the Study……………………………..……………....................1-2

Statement of the Problem……………………………………………….……...….2

Hypothesis…………………………………….………………………….………..3

Significance of the Study……………………………...……..…………...…..…3-4

Scope and Delimitations………………………………………………….…..…...4

Definition of Terms…………………………………………….…………….…4-7

Chapter 2: REVIEW OF RELATED STUDIES AND LITERATURE

Related Literature…………...……………………………….…...…..….….…8-13

Related Studies……………………………………………………….………13-16

Conceptual Framework……………...……………..………………..…….…17-18

Chapter 3: METHODOLOGY

Research Design………………………..…………………………..……..….19-20

Materials/Equipment……………………………………………..………………20

Preparation of the study animals…………………………………….……….20-21

Procedure……………………………………...………………………...…….…21

Disposal of Materials……………………...………………………....…………22

Risk and Safety Procedure………………………………………………………22

Pros and Cons………………………………………………………...………….22

 Collection of Data…………………………………….…………………………23

Data Analysis……………………………………………………..……..……….23

Chapter 4: RESULTS AND DISCUSSIONS

Results and Discussions………………………………………………………24-28

Chapter 5: SUMMARY, CONCLUSIONS AND RECOMMENDATIONS

Summary…………………………………………………………………………29

Conclusions………………………………………………………………………30

Recommendations……………………………………………………..……..30-31

BIBLIOGRAPHY……………………………………………………………………32-33
APPENDICES
A Plate ………………………..……………………………….….…..34-35
B Tables…………………………………………………………………36-37
C Statistical Computations ………..….………………………...…..…..38-39
CURRICULUM VITAE…………………………………………………………………40
 ABSTRACT

Title: Cognitive Ability Assessment of Albino Rats (Rattus albus) Before and After

Exposing it to Vaporized Electronic Cigarette Juice (Nicotine-Free).

Proponent: Mark kjil C. Tayaba

School: Calasiao Comprehensive National High School

Electronic Cigarette juice is one of the addictive substances that most of the

teenagers use it on their electronic cigarette. This study aimed to determine the effects of

vaporized Electronic Cigarette juice(Nicotine-Free) in the cognitive ability of the Albino

rats in terms of Time taken to find the platform, speed and circling information. In this

study, the researchers focused on the effects of vaporized electronic cigarette

juice(Nicotine-Free) to albino rats before and after exposing to Vaporized Electronic

Cigarette Juice (Nicotine-Free). Instead of using the electronic cigarette, the researchers

used a nebulizer for vaporizing the electronic cigarette juice.

The Albino rats were observed (for 7 days with one day interval) after exposing it

in the Vaporized Electronic Cigarette Juice (Nicotine-Free) . Thus, the researcher

observed the time it took to find the platform , speed and circling information of the

Albino rats.

T-test declared that the means of the Pretest and Posttest are significant at .05

level of significance. It declared a significant difference among the Pretest and Posttest in

terms of time taken to find the platform , speed and circling information . F-test further

declared that the means of Pretest and Posttest are significant difference. It can be
concluded that the duration of exposure to Vaporized Electronic Cigarette Juice

(Nicotine-Free) has an effect to the Cognitive Ability of the Albino rats.

i
 Acknowledgement

This research study was supported by several individuals who shared their

ideas to complete this study. The researcher would like to thank everyone, especially to

the following individuals listed below who made this study possible.

To Our Almighty God, for giving the researchers courage and guidance to finish

this study.

To Ms. Geraldine Ocampo their research adviser, for giving pieces of advice to

improve the manuscript of this study and for guiding the researchers.

Also, the researcher families for giving sufficient financial and moral support.

Without them, the researcher would not be able to conduct this study.

And to all the people who help the researcher to finish this, who are not

mentioned thank you very much.

ii
 Chapter 1

THE PROBLEM: RATIONALE AND ITS BACKGROUND

Background of the Study

Electronic cigarette is a equipment that is used to reproduce the experience

of smoking, having a cartridge with a heater that terminates liquid nicotine or electronic

cigarette juice instead of burning tobacco. E- cigarette smoking is extensively used

throughout the world especially teens because of the juice that is used - it is fragrant and

has different flavors to choose from (Cheng, 2014). Electronic cigarette juice come in

sweet flavors, like fruit, candy, and dessert. E-cigarette juices are tobacco products that

carry nicotine. Nicotine is an extremely addictive substance, and several of today’s youth

who are using Electronic cigarettes could develop into tomorrow’s cigarette smokers

(Tracy, 2015). Nicotine’s hazard can also damage brain improvement in procedures that

may disturb the health of kids. The brain of youth and young adults is more accessible to

the negative results of nicotine vulnerability. The effects include addiction, motivate for

use of other addictive materials, decreased impulse control, loss in attention and

acknowledgment, and mood disorders (Grana, 2014).

Meanwhile, the Albino rats are one of the animals used in most laboratory test

and research because it is believed that these albino rats have co-evolved with humans for

many decades (Hedrich, 2010) The Albino rats consume 76% of the day sleeping and

24% of the time awake making them nocturnal animals which they have tended to sleep

during the day and stake awake at night (Raedler et al., 2015).
 2

In addition, Albino rats are one of the most commonly used animals in biology

and in research because rats are small and are also easy to be handled. They are also

intelligent, cooperative in social activities, and have valuable memories especially for

odor and flavor.

So, the researcher will conduct this study to determine the effects of the vapor of

Electronic juice in the cognitive ability of the Albino rats. It will also help educate the

persons who are always using Electronic cigarette or exposed in Electronic cigarette juice

vapor on its effects.

Statement of the Problem

Generally, the aim of this study is to determine the cognitive ability assessment of

Albino Rats upon exposure to Vaporized Electronic Cigarette Juice (Nicotine-Free).

Specifically, this study sought to answer the following question:

1. Is there significant difference in cognitive ability of albino rats before and

after exposing it to Vaporized Electronic Cigarette Juice (Nicotine-Free)

in terms of :

a) time taken to find the platform;

b) speed; and

c) circling information.
 3

Hypothesis

This study was test the following hypothesis with the probability level of 0.05.

1. There is no significant difference in the cognitive ability of albino rats before

and after exposing it to Vaporized Electronic Cigarette Juice (Nicotine-Free)

in terms of:

a) time taken to find the platform;

b) speed; and

c) Circling information.

Significance of the Study

This study will benefit the following:

Families

This study is beneficial to the parents whose children or even them who use

Electronic Cigarette, this study will help them gain more knowledge about the ill effects

of having been exposed continuously to Electronic Cigarette Juice.

Smokers

This study is beneficial to the people who smoke Electronic cigarette, it will help

them to gain more knowledge about the things that will possibly happen to them if they

continuously smoke Electronic cigarette. This study will also be beneficial to the people

who experienced second hand smoke, it will help them to understand what will happen

them if they are exposed to electronic cigarette juice vapor even in a short span of time.
 4

Companies

This study will also be beneficial for the companies of Electronic cigarette juice

such as the Humble Juice Company and the Beard Vape Corporation , to know more

ideas in reducing the use of chemicals that are harmful and the effects of their products to

the people and give necessary precautions to the resellers of their product that they will

produce. This study will also benefits the Agencies and Organizations that are against in

Electronic cigarette smoking such as the Food and Drug Administration (FDA); it will

help them to gain more knowledge and ideas for the precautions of the tobacco products

that are going through to their Agencies before releasing it in the market.

Scope and Delimitation

This study has One-Group Pretest-Posttest. The Pretest-Posttest are compared and

evaluated by observing the Speed, Time it takes to find the platform and circling

information of the Albino rats, six male Albino rats was used in this study, it was tested

by using the morris water test because it is commonly used in testing the Cognitive

Ability of the Albino Rats. This study was conducted at the JICA Laboratory at Calasiao

Comprehensive National High School at the month of August to October 2019. The

Electronic cigarette is replaced by a Nebulizer for the researcher safety. This study focus

on the effects of exposure to vaporized Electronic Cigarette Juice (Nicotine-Free). in the

Cognitive Ability of the Albino Rats.

 5

Definition of terms

For further understanding, the following terms were defined.

Assessment. Is refers to the wide variety of methods or tools that educators use to

evaluate, measure, and document the academic readiness, learning progress, skill

acquisition, or educational needs of students ( Greenberg, 2000).

Albino Rat .Is a small mammal that is also called the Pink-Eyed Whites. A type

of mice that is usually used in the laboratory and for scientific research. The first

mutation that is discovered and knowingly to bred. These rats are also kept, bred and are

sold to the market as pets and for scientific research (Hedrich, 2010). The Experimental

unit of this researchers study.

Circling information .A circling information is a simple close shape. It is the set

of all points in a plane that are at a given distance from a given point, the center;

equivalently it is the curve traced out by a point that moves so that its distance from a

given point is constant. The distance between any of the points and the center is called

the radius (Hazewinkel, 2001), Circling information of rat that was tested in this study,

and this refers to the number of circles made by the test animal before reaching the

platform in the Morris water test, used as determinant of the cognitive ability of the

albino rats.

Cognitive Ability. Is an intellectual process and it is also called "intelligence" or

"general mental ability." of humans, which is marked by complex cognitive feats and

high levels of motivation and self-awareness. Through their intelligence, humans possess

the cognitive abilities to learn, form concepts, understand, apply logic, and reason,
 6

including the capacities to recognize patterns, comprehend ideas, plan, solve problems.

(Johnson, 2010), Cognitive Ability of rat was tested in this study,through Morris Water

Test

Electronic Cigarette Juice. E-juice or vape juice is the fluid that is used in and

personal electronic cigarettes and vaporizers that creates an actual vapor. It contains

nicotine but it can also be made without nicotine depending on personal preference. The

Electronic cigarette heats up the Electronic cigarette juice between 200-400 degrees

Fahrenheit which turns it into vapor when you inhale. Electronic cigarette juice or vape

juice is a mixture of food grade flavoring, vegetable glycerin (VG) or waterpropylene

glycol (PG), and a choice of nicotine levels or zero nicotine. It is safe to consume and

often, quite delicious (Biovotion, 2018). One of the materials that was used by the

researchers.

Nicotine.A chemical that contains nitrogen and alkaloid.It is containsby some

types of plants, and one is the tobacco plant. It is also produced synthetically. The

tobacco plant is a very common plant that is a main component of a product that has

nicotine. Nicotine is really addictive and if you are exposed frequently, it can cause some

complications (Weatherspoon, 2018).

Morris Water Test. It is widely used in behavioral neuroscience to study spatial

learning and memory. It enables learning, memory, and spatial working to be studied

with great accuracy, and can also be used to assess damage to particular cortical regions

of the brain, It is used by neuroscientists to measure the effect of neurocognitive

disorders on spatial learning and possible neural treatments, to test the effect to the brain
 7

in areas concerned with memory, and to study how age influences cognitive function and

spatial learning.(Hunter, 2012) Morris water test was used to test the cognitive ability of

the albino rats.

Speed.Is the magnitude of its velocity (the rate of change of its position); it is thus

a scalar quantity. The average speed of an object in an interval of time is

the distance travelled by the object divided by the duration of the interval; the

instantaneous speed is the limit of the average speed as the duration of the time interval

approaches zero (Wilson, 2002). Speed was obtained by dividing the distance and the

time it took for the Albino rats to find the platform

Vapor. A substance in the gas phase that is a temperature and lower than its

critical temperature. It can also exist even if it’s in a state of a solid or a liquid substance.

It is responsible for the process of cloud formation and condensation (Cheng, 2014). The

different group that were used in the study was tested at the Albino rats in the form of

vapor.
 Chapter 2

REVIEW OF RELATED LITERATURE AND STUDIES

Albino Rats

Albino rats are also called PEW it means Pink-Eyed White, a typical name for

albino rats among fanciers and raisers. Albino rats spend the vast majority of the daytime

hours sleeping. The pink of an albino rat’s eyes isn't real eye shading. An albino rat

creature has no pigmentation in its hair or eyes, so the pink that you see isn't eye shading.

What you see is the shade of the veins inside the rodent's eyes. Just unadulterated white

rats with pink eyes can be delegated albino. The official PEW standard calls for splendid

white hide with no trace of yellow, and medium pink eyes. a genuine albino rat and not

only a light-shaded extravagant rodent. Particularly Albino rats, have feeble visual

perception, they utilize this influencing development to enable them to identify

movement in their condition (Vandenburgh, 2016). The rat found early use in laboratory

research in five areas: W. S. Small suggested that the rate of learning could be measured

by rats in a maze; a suggestion employed by John B. Watson for his Ph.D. dissertation in

1903. The first rat colony in America used for nutrition research was started in January

1908 by Elmer McCollum and then nutritive requirements of rats were used by Thomas

Burr Osborne and Lafayette Mendel to determine the details of protein nutrition.

The reproductive function of rats was studied at Institute for Experimental Biology

at University of California, Berkeley by Herbert McLean Evans and Joseph A.

Long.[6] The genetics of rats was studied by William Ernest Castle at the Bussey
 9

Institute of Harvard University until it closed in 1994. And rats have long been used

in cancer research; for instance at Crocker Institute for Cancer Research

Indeed, rats have additionally co-advanced with people for many years. They eat a

large portion of indistinguishable nourishments from we do and live in the majority of

similar spots. They can get huge numbers of indistinguishable illnesses from we do.

Organically, they're fundamentally the same as us, which makes results from their tests

exchange so almost to us (Charney, 2016).

Cognitive Ability

Cognitive is relating to involve conscious intellectual activity such as thinking,

reasoning or remembering. Cognitive is based on or capable of being reduced to

empirical factual knowledge; Cognitive functioning is a term referring to an individual’s

ability to process to (thoughts) that should not deplete on a large scale in healthy

individuals. It is defined as the ability of an individual to perform the various mental

activities most closely associated with learning and problem solving. Example includes

verbal, spatial, psychomotor, and processing-speed ability. Cognition mainly refers to

things like memory, the ability to learn new information, speech, and understanding of

written material. The brain is usually capable of learning new skills in the aforementioned

areas, typically in early childhood, and of developing personal thoughts and beliefs about

the world. Old age and disease may affect cognitive function, causing memory loss and

trouble thinking of the right words while speaking or writing, for example, can eventually

cause memory loss, an inability to grasp new concepts of information, and depleted
 10

verbal fluency. Not all with the condition will experience this side effect, and most will

retain their general intellect and the ability (Weinberg, 2012).

Humans generally have a capacity for cognitive function once born, the ability of

an individual to perform the various mental activities most closely associated with

learning and problem solving. Examples include verbal, spatial, psychomotor, and

processing-speed ability so almost every person is capable of learning or remembering.

However, this is tested using tests like the IQ test, although these have issues with

accuracy and completeness. In these tests, the patient will be asked a series of questions

or to perform tasks, with each measuring a cognitive skill, such as level of consciousness,

memory, awareness, problem-solving, motor skills, analytical abilities, or other similar

concepts ]Early childhood is when most people are best able to absorb and use new

information. In this period, children learn new words, concepts, and various methods to

express themselves. (Weinberg, 2012).

Electronic Cigarette

Electronic cigarettes, are battery-operated devices that were initially shaped like

cigarettes, but now include vape mods, Juuls, and vape pens. The brand-name products

contain nicotine, an addictive drug that stimulates, relaxes, and is naturally found in

tobacco. It is the nicotine in cigarettes that makes smoking so addictive, and the same is

true for most vaping and juuling. These electronic products allow nicotine to be inhaled,

and they work by heating a liquid cartridge containing nicotine, flavors, and other

chemicals into a vapor. Because e-cigarettes heat a liquid instead of tobacco, what is

released is considered smokeless. also known as vape pens, e-hookahs, hookah pens, or
 11

vape pipes, are electronic nicotine delivery systems, invented in the early 2000s in China

and introduced recently to the US market. They consist of a battery, an atomizer with a

heating element, and a cartridge containing nicotine and flavoring. When heated, the

device produces an aerosol, also called a vapor by marketers. This has led to the use of

the term “vaping” when using them, rather than smoke. The brands manufactured by

Imperial Tobacco and RJ Reynolds look very similar to cigarettes, while the refillable

types look very different. E-cigarettes are widely available both via the internet and retail

settings including convenience stores where adolescents are particularly likely to

frequent—about two-thirds of youth shop at convenience stores at least once a week. In

early 2014, there were 466 brands and 7764 unique flavors of e-cigarette products. Some

of these flavors mimic candy – examples are Bubble Gum, Snicker Doodle, Gummy

Bear, Mountain Dew, Captain Crunch, and Skittles. (Bostean, 2016)

E-cigarettes come in a variety of forms and include vape mods, Juuls, and vape

pens. There are brand name products (Juul is the most widely used) and “home-made”

versions. Some contain high levels of nicotine, while others contain marijuana or just

contain flavoring. The focus of this article is on e-cigarettes because most of the research

that exists has been done on them, but much of the information below is relevant to these

other products as well. n general, they contain much lower levels of the harmful

constituents present in conventional cigarettes. However, they do contain nicotine, an

addictive and powerful psychoactive substance that acts in the brain and throughout the

body. Little internal quality control is present in the manufacture of e-cigarettes and

currently, there is no external regulatory oversight of ingredients or packaging. (Evans,

2017). Thus, there is wide variability in e-cigarette device engineering as well as

 12

differences in the nicotine concentrations of the 2 solutions (called e-liquids) that go into

the device. The nicotine level listed on the labels of cartridges and solutions often differ

significantly from measured values. Several studies have scientifically evaluated contents

of the e-cigarette aerosol. One study analyzed the aerosol from 12 brands of e-cigarettes

and compared the contents to those of a nicotine inhaler. Although toxic substances were

found, they were 9-450 times lower than in cigarette smoke and were comparable in

some cases to the nicotine inhaler. Another recent study also found that the toxic contents

of e-cigarette aerosol differed greatly among brands. This variability in the contents of e-

cigarette aerosol is a concern and has prompted the World Health Organization (WHO) to

conclude that the regulation of e-cigarettes is a necessary precondition for the public to

have reliable information on the potential risks and benefits of using e-cigarettes (Grana,

2014)

In fact, a preliminary study presented at the 2018 annual meeting of the American

Chemical Society found that vaping could damage DNA [8]. The study examined the

saliva of 5 adults before and after a 15-minute vaping session. The saliva had an increase

in potentially dangerous chemicals, such as formaldehyde and acrolein. Acrolein has

been proven to be associated with DNA damage, for example, and DNA damage can

eventually cause cancer.

Because they are smokeless, many incorrectly assume that e-cigarettes are safer for non-

smokers and the environment than traditional cigarettes. However, a study published in

the International Journal of Hygiene and Environmental Health found that the use of e-

cigarettes results in increased concentrations of volatile organic compounds (VOCs) and

airborne particles, both of which are potentially harmful when inhaled.[11] Although e-
 13

cigarette vapor may not result in the obvious smell and visible smoke of traditional

cigarettes, it still has a negative impact on air quality, especially when vaping indoors

Related Studies

Albino Rats

The utilization of rats in research centers has establishes somewhere down ever,

extending back a few hundred years. Rats are a characteristic decision for lab testing for a

few reasons: They're profoundly astute and, when restrained, are cordial and resigned.

Rats are simple to think that they have short life span which makes them perfect for

concentrate some restorative subjects. Rats are communal animals. Young are raised

communally with shared nursing responsibilities. Males can be co-housed as fighting

rarely occurs among adult they are burrowers and take advantage of that whenever the

opportunity presented. They are nocturnal animal and can easily adapt to their

environments. Rats can inflict severe bite wounds and must be handled gently and with

care to avoid biting (Johnson, 2010).

Albino male rats of a pure strain, inbred in this laboratory for 3 years were

employed. The basal diet consisted of dog chow ad lib., supplemented with greens.

Hypophysectomy was accomplished by the technic of Richter and Wislocki with the

exception that a tracheotomy, as suggested by Fran-seen, (3) was usually performed.

Success of the operation was based upon failure of weight to increase, atrophy of testes,

atrophy of thymus, atrophy of cortex of adrenal and absence of visible hypophyseal tissue

at autopsy. While the completeness of hypophysectomy was not checked histologically

in- these experiments, similar studies have been performed elsewhere (at Cornell
 14

University Medical College) by one of us (R. A.P.) on Wistar strain rats. The chemical

analyses reported in this paper were verified on the fasting animal and on the animal 3

hours after the administration of glucose. (Mordes, 2004)

A 1972 study compared neoplasms in Sprague Dawley rats from six different

commercial suppliers and found highly significant differences in the incidences

of endocrine and mammary tumors. There were even significant variations in the

incidences of adrenal medulla tumors among rats from the same source raised in different

laboratories. All but one of the testicular tumors occurred in the rats from a single

supplier. The researchers found that the incidence of tumors in Sprague-Dawley rats from

different commercial sources varied as much from each other as from the other strains of

rats. The authors of the study "stressed the need for extreme caution in evaluation

of carcinogenicity studies conducted at different laboratories and/or on rats from different

sources.(Krinke, 2000)

Electronic Cigarette

E-cigarettes are a relatively new tobacco product that have been sold in the U.S.

for about a decade The e-cigarettes currently in the U.S. marketplace have not been

systemically reviewed by the Food and Drug Administration to determine their impact on

lung health. While much remains to be determined about the lasting health consequences

of these products, the American Lung Association is very troubled by the evolving

evidence about the impact of e-cigarettes on the lungs.

 15

In January 2018, the National Academies of Science, Engineering and

Medicine1released a consensus study report that reviewed over 800 different studies.That

report made clear: using e-cigarettes causes health risks. It concluded that e-cigarettes

both contain and emit a number of potentially toxic substances. The Academies' report

also states there is moderate evidence that youth who use e-cigarettes are at increased risk

for cough and wheezing and an increase in asthma exacerbations. A study from the

University of North Carolina found that the two primary ingredients found in e-cigarettes

propylene glycol and vegetable glycerin are toxic to cells and that the more ingredients in

an e-liquid, the greater the toxicity. E-cigarettes produce a number of dangerous

chemicals including acetaldehyde, acrolein, and formaldehyde. These aldehydes can

cause lung disease, as well as cardiovascular (heart) disease ( Sassano, 2018).

In 2016, the Surgeon General concluded that secondhand emissions contain,

"nicotine; ultrafine particles; flavorings such as diacetyl, a chemical linked to serious

lung disease; volatile organic compounds such as benzene, which is found in car exhaust;

and heavy metals, such as nickel, tin, and lead (Ogunwale,, 2016).

Since 2009, FDA has pointed out that e-cigarettes contain “detectable levels of

known carcinogens and toxic chemicals to which users could be exposed.” For example,

in e-cigarette cartridges marketed as “tobacco-free,” the FDA detected a toxic compound

found in antifreeze, tobacco-specific compounds that have been shown to cause cancer in

humans, and other toxic tobacco-specific impurities.[4] Another study looked at 42 of

these liquid cartridges and determined that they contained formaldehyde, a chemical

known to cause cancer in humans.[5] Formaldehyde was found in several of the cartridges

at levels much higher than the maximum EPA recommends for humans. In 2017, a study
 16

published in the Public Library of Science Journal showed that significant levels of

benzene, a well-known carcinogen, were found in the vapor produced by several popular

brands of e-cigarettes. (Ogunwale,, 2016).

 17

Conceptual Framework

A conceptual framework represents the researcher synthesis of literature on how

to explain a phenomenon. It maps out the actions required in the course of

the study given his previous knowledge of other researcher point of view and his

observations on the subject of research

To understand more about the factors involved in the study, the researcher

illustrated the diagram below:

Exposure of Vaporized Electronic Cigarette

Juice in Albino Rats.

Independent Variable

Temperature

Effects on Albino Rats’ Cognitive

Ability (a) time taken to find the Intervening Variable
platform, (b) speed (c) circling
information (number of cirlces),

Dependent Variable

Figure 1. Paradigm of the Independent Variable and Dependent Variable of the study
“Cognitive Ability Assessment of Albino Rats (Rattus albus) Before and After
Exposing it to Vaporized Electronic Cigarette Juice (Nicotine-Free).”
 18

The independent variable was the Exposure of Vaporized Electronic Cigarette

Juice in Albino Rats that was the variable that affect the condition of the rats and was

also controlled by the researcher. The dependent variable is Effects on Albino Rats’

Cognitive Ability (a) time taken to find the platform, (b) speed (c) circling information

(number of cirlces) these is the variable that the independent variable will alter. The

intervening variable is the daily Temperature. This was not be controlled or affected on

the experimentation process.

 Chapter 3

METHODOLOGY

Research Design

A One-Group Pretest-Posttest design was used in this study because it

characterized by two features which were the Pretest and Posttest. The Posttest was

compared to the parameters which where the time it take to find the platform, speed and

circling information of the Pretest. One-Group Pretest-Posttest design is suitable because

there are comparable which are before and after.

Table 1.1 One-Group Pretest-Posttest design for the time (seconds) it took before finally
the platform.

Subjects Pretest Posttest

1 x X
2 x X
3 x X
4 x X
5 x X
6 x X
Mean x X

Table 1.2 One-Group Pretest-Posttest design for the speed (m/s) of the Albino rats before
finally found the platform.

Subjects Pretest Posttest

1 x X
2 x X
3 x X
4 x X
5 x X
6 x X
Mean X X
 20

Table 1.3 One-Group Pretest-Posttest design for the Circling Information of the Albino
rats before finally found the platform.

Subjects Pretest Posttest

1 x X
2 x X
3 x X
4 x X
5 x X
6 x X
Mean X X

Materials/Equipment

The materials that were used in the study were one airtight container which was

bought at Dominisac in Calasiao, Pangasinan. 93 mL of Electronic cigarette juice were

bought by the researchers accompanied by a guardian at Cloudline, a vaping station in

Calasiao, Pangasinan. One improvised Morrris Water Test, and two pairs of gloves and a

gas mask was also used for the safety of the researcher during the experimental period. A

nebulizer found at the researcher house will be used to vaporize the electronic cigarette

juice in this study.

Preparation of Study Animals

Four male albino rats about 9 weeks old approximately of the same weight were

acquired from Frankie’s pet shop in San Carlos City under Bureau of Animal Industry

regulation ( BAI Registration No. PTS-0031). The albino rats were housed and given

access to food and water. The albino rats were acclimatized in the testing site for seven

days prior to experimentation. All the animals used in the experiment were treated as
 21

humanely as possible according to the specifications and guidelines set by the

Department of Basic Sciences College of Veterinary Medicine Benguet State University.

Collection and Preparation of the Electronic Cigarette Juice

One bottle of electronic cigarette juice (Nicotine-Free) was utilized in the study

that was bought from Cloudline in Calasiao, Pangasinan. The researcher were

accompanied by a guardian in buying the electronic cigarette juice.

Procedure

A. Preparation of Morris Water Test.

Prior to the experiment proper, the researcher first prepared the device that

was used to test cognitive ability. A plastic water tub usually used at home for

washing clothes was filled with about 3 gallons of water. A platform which the

Albino rats will swam for and eventually sat on was situated at the center of the

plastic water tub.

B. Exposure of Study Animals to Electronic Cigarette Juice.

Six Albino rats was utilized as test animals and was bought at Frankie’s

pet shop in San Carlos City, Pangasinan.. The researchers had one group, pretest

and posttest. Posttest were exposed to the Vaporized Electronic Cigarette Juice

was prepared by the researcher. The time it took for them to find the platform,

their speed and the number of circles they have to travel to reach the platform was

recorded.
 22

Disposal of Materials

After the experiment, the container of the electronic cigarette juice and the hose of

the nebulizer used in the experiment were put inside a plastic bag and the researcher was

tie the edges of the plastic bag for security. The researcher threw the plastic bag

containing the container of electronic cigarette juice directly on a trash bin. On the other

hand, the 4 Albino rats were allowed to recover from the experiment and will be used for

the next experiment.

Risk and Safety Procedures

In conducting the experiment, the researcher used gloves in handling the Albino

rats. The researcher also read some guidelines about proper handling of rats. Carefulness

was also needed in handling the Albino rats so that there will be a low chance of getting

bitten.

Pros and Cons

The researcher was in handling the Albino rats. The researcher used gloves while

handling the Albino rats. This will help to avoid to get bitten by the Albino rats. On the

other hand, the researcher threw the container of the electronic cigarette juice and the

hose attached to the nebulizer. The container of the electronic cigarette and the hose of

the nebulizer were placed inside a plastic bag was secured by tying the edges of the

plastic bag. The researchers threw the plastic bag containing the container of electronic

cigarette juice directly on a trash bin, despite the precautions. This research aimed to

determine the effects of the vapor of Electronic cigarette juice in Albino rats.
 23

Data Collection

The seven-day test in assessing the cognitive ability of Albino rats after exposure

to Vaporized Electronic Cigarette Juice was determined in the following:

a) Time taken for each albino rat to find the platform after being hidden in the

plastic water tub. This was done by observing and recording the number of

seconds it took them to find the platform;

b) Speed was determined by the distance traveled and time it took for the albino

rats to find the platform and

c) Counting the number of circles they have to go about before finally reaching

the platform.

Data Analysis

T-test was the method used for analyzing data to determine if Vaporized

Electronic Cigarette (Nicotine-Free). The researcher wants to know the significant

difference between the Pretest and Posttest.

 Chapter 4

RESULTS AND DISCUSSION

This chapter presents all results and findings after the conducted experiments.

This research study tested the cognitive ability of the albino rats before and after

exposing it to Vaporized Electronic Cigarette Juice (Nicotine-Free). Specifically, this

study sought to answer the following question: 1) Is there significant difference in

cognitive ability of Albino rats before and after exposing it to vaporized electronic

cigarette juice (Nicotine-Free) in terms of : a) time taken to find the platform; b) speed;

and c) number of circles

One-Group Pretest-Posttest design was used in this study. Test animals that was

used in this study. Six albino rats was used in this study. Before the exposure to the

Vaporized Electronic Cigarette Juice (Nicotine-Free) they were first observed to the

morris water test after that they were exposed to the Vaporized Electronic Cigarette Juice

(Nicotine-Free). The rats were observed in the following: the time it took for them to

find the platform, their speed and their circling information, i.e. the number of circles

formed before landing themselves on the platform.

 25

The tables on the next pages show the data gathered from the test parameters of

the study.

a. Time taken to find the platform

Time taken for each albino rat before and after found the platform and was
observed and recorded.
Table 3.1 Average Time (in seconds) Before and After Taken by the Albino Rats in
Finding the platform.

Mean
Pretest 19.22 seconds

Posttest 33.45 seconds

The table shows the means of Pretest and Posttest, Posttest obtained the longest

time (33.45 seconds) to find the platform in the plastic water tub as compared to the

Pretest . The T-test on the mean of time taken by the albino rats to find the platform

before and after exposure to Vaporized Electronic Cigarette Juice (Nicotine-Free)

declared that a statistical significance among the treatments was evident at 0.05

significance level. This further suggests that as exposure to Vaporized Electronic

Cigarette Juice (Nicotine-Free) becomes longer, the greater the struggle to complete

Morris water test for the Albino rats. Similarly, in the study of Slikker (2000) on the

effects of neurotoxins on cognitive ability groups exposed to neurotoxin chemicals

consistently have deficits on cognitive tests based on performance. Hence that Electronic

Cigarette Juice (Nicotine-Free) chemicals can be neurotoxins because based on their

performance there is an effect on their cognitive ability.

 26

Tme taken to find the platform (seconds)

40

30

20

10

0
Pretest Posttest
Tme taken to find the platform (seconds)

Figure 2.1 Average mean of time finding the platform (seconds) before and after
exposing it to Vaporized Electronic Cigarette Juice (Nicotine-Free).

b. Speed

Speed is determined by determining the distance traveled and the time it took for
the albino rats to find the platform. This is in term of m/s.
Table 3.2. Average speed of the albino rats before and after exposing it to Vaporized
Electronic Cigarette Juice (Nicotine-Free).

Mean
Pretest 10.22 m/s

Posttest 17.80 m/s

Calculated T-test on the means of speed taken by the albino rats to find the

platform between the Pretest and Posttest declared that a statistical significance is

observed among the four groups. The table above shows that posttest obtained an

average mean of 17.80 m/s to be the fastest among Posttest and Pretest.
 27

Speed (m/s)
20

15

10

0
Pretest Posttest
Speed (m/s)

Figure 3.1 Comparison of speed of the albino rats before and after finally reaching the
platform.

c. Circling information

Below is the table showing the number of circles made travelled by the albino rats

before landing themselves in the platform.

Table 3.3 Average circles made by the albino rats before and after reaching the platform

Mean
Pretest 1.10

Posttest 2.41

The T-test on the average circles made by the albino rats before reaching the

platform in Morris water test between the Pretest and Posttest declared that a statistical

significance is observed among the Posttest and Pretest. The table above shows that the

Posttest obtained a circling average of 2.41 ranking the test animals in this group to be

the fastest to reach the platform.

 28

Circling information
3
2.5
2
1.5
1
0.5
0
Pretest Posttest

Figure 4.1 Comparison of number of circle before and finally reaching the platform.
 Chapter 5

SUMMARY, CONCLUSIONS, AND RECOMMENDATIONS

Summary

This research study aimed to determine the cognitive ability of albino rats before

and after expose to Vaporized Electronic Cigarette Juice (Nicotine-Free). Specifically,

the study sought to answer the following question 1) Is there significant difference in

cognitive ability of Albino rats before and after exposing it to Vaporized Electronic

Cigarette Juice (Nicotine-Free) in terms of : a) time taken to find the platform; b) speed;

and c) circling information. One-Group Pretest-Posttest design was used in this study.

Six albino rats Test animals was used in this study which were first not expose to

Vaporized Electronic Cigarette Juice (Nicotine-Free) served as Pretest. After the

exposure of Vaporized Electronic cigarette Juice (Nicotine-Free) served as the Posttest.

Test animals in the Pretest and Posttest was tested using Morris water test. The rats

were observed in the following: the time it took for them to find the platform, their speed

and their circling information, i.e. the number of circles formed before landing

themselves in the platform. The gathered data were subjected to statistical analysis and

were interpreted an analyzed.

The following are the salient features of the study:

1. The Cognitive ability of the Albino rats before and after exposed to Vaporized

Electronic Cigarette Juice (Nicotine-Free) have deficits on cognitive tests based

on performance as manifested in the results of the time it took for the albino rats

to find the platform and their circling information.

 30

2. The Cognitive ability of the Albino rats before and after exposed to Vaporized

Electronic Cigarette Juice (Nicotine-Free) have deficits on cognitive tests based

on performance as manifested in the results of the speed (m/s) of the albino rats

to find the platform and their circling information.

3. The Cognitive ability of the Albino rats before and after exposed to Vaporized

Electronic Cigarette Juice (Nicotine-Free) have deficits on cognitive tests based

on performance as manifested in the results of the circling information of the

albino rats to find the platform and their circling information.

Conclusions

Based on the findings of the study the following conclusions were drawn:

1. As exposure Vaporized Electronic Cigarette Juice (Nicotine-Free) becomes

longer, the greater the struggle to complete Morris water test for the Albino rats.

Evidently, between the Pretest and Posttest, those test animals which were

exposed to Vaporized Electronic Cigarette Juice (Nicotine-Free) had the longer

time finding the platform.

Recommendations

Based on the conclusions of the study, the following recommendations

were formulated:

1. Changes in weight of the Albino rats after the exposure to Electronic

Cigarette Juice (Nicotine-Free) be investigated.

2. Further research on how Electronic Cigarette Juice (Nicotine-Free) of affects

social and grooming skill of the Albino rats.

 31

3. Lastly, the researcher recommends to inspect and observe the lungs of the

Albino rats after exposure to Electronic Cigarette Juice (Nicotine-Free)

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 34

APPENDIX A

Plates

Juice (Nicotine-Free)
Gloves

Water Tub

Nebulizer

Plate 1. Materials that were used in the study entitled “Cognitive Ability Assessment of
Albino Rats (Rattus albus) Before and After Exposing it to Vaporized Electronic
Cigarette Juice (Nicotine-Free).”

Plate 2. Water maze used in the study entitled “Cognitive Ability Assessment of Albino
Rats (Rattus albus) Before and After Exposing it to Vaporized Electronic Cigarette Juice
(Nicotine-Free).”
 35

Plate 3. Albino rats that were used as an experimental unit in the study entitled
“Cognitive Ability Assessment of Albino Rats (Rattus albus) Before and After Exposing
it to Vaporized Electronic Cigarette Juice (Nicotine-Free).”

Plate 4. Albino rats while doing the experiment on the study entitled “Cognitive Ability
Assessment of Albino Rats (Rattus albus) Before and After Exposing it to Vaporized
Electronic Cigarette Juice (Nicotine-Free).”
 36

APPENDIX B

Tables

Table B.1. F-test result for the time before and after taken to find the platform rats

Statistical Analysis

F-value: 0.34 F-critical 0.59

P-value: 0.0003 Alpha:0.05

Table B.2. F-test result for the Speed of the Albino rats.

Statistical Analysis

F-value: 0.34 F-critical: 0.59

P-value: 0.0003 Alpha:0.05

 37

Table B.3. F-test result for the Circling information of the Albino rats.

Statistical Analysis

F-value: 1.31 F-critical: 1.69

P-value: 0.19 Alpha:0.05

 38

APPENDIX C

Statistical Analysis

SUMMARY

Statistical Computation T-test: Paired Two Sample Time taken to find the platform

19.84 43.38
Mean 19.217561 33.45146341
Variance 31.0318289 90.1433478
Observations 41 41
Pearson Correlation 0.41565679
Hypothesized Mean
Difference 0
df 40
t Stat -10.372578
P(T<=t) one-tail 3.3272E-13
t Critical one-tail 1.68385101
P(T<=t) two-tail 6.6544E-13
t Critical two-tail 2.02107537

SUMMARY

Statistical Computation T-test for the Speed of albino rats.

10.55543733 23.07937859
Mean 10.22428228 17.79711822
Variance 8.783679875 25.51542523
Observations 41 41
Pearson Correlation 0.415656788
Hypothesized Mean
Difference 0
df 40
t Stat -10.37257788
P(T<=t) one-tail 3.32721E-13
t Critical one-tail 1.683851014
P(T<=t) two-tail 6.65443E-13
t Critical two-tail 2.02107537
 39

SUMMARY

Statistical Computation A.I.ANOVA Number of Circles Travelled by albino rats.

2 5
Mean 1.097560976 2.414634146
Variance 0.790243902 0.598780488
Observations 41 41
Pearson Correlation 0.303157182
Hypothesized Mean
Difference 0
df 40
t Stat -8.55420388
P(T<=t) one-tail 7.1221E-11
t Critical one-tail 1.683851014
P(T<=t) two-tail 1.42442E-10
t Critical two-tail 2.02107537
 40

CURRICULUM VITAE

Mark kjil C. Tayaba

#333 Brgy. Bacayao Sur Dagupan City, Pangasinan

markkjil15@gmail.com

09566327532

------------------------------------------------------------------------------------------------------------

PERSONAL INFORMATION

Age: 15 years old Favorite Color: Pink

Birthdate: April 15, 2004 Ambition: Accountancy

Civil Status: Single Favorite Song: When we were young

Gender: Male Hobby: Listening to music, watvhing

vlogs

Nationality: Filipino

Relegion: Catholic

Motto: “Time is gold.”

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EDUCATIONAL BACKGROUND

Junior Highschool – Calasiao Comprehensive National Highschool

Calasiao, Pangasinan

2019 - 2020

Primary Education – Bacayao Sur Elementary School

Dagupan City

2015 - 2016