A Research Proposal Titled

Antibacterial Activity of Passion fruit (Passiflora sp.) Leaf Extracts

Jaslynn Ross B. Pinto

Matthiea Quinella Deonne N. Angobung

Dell Wydlov A. Taccad

In Partial fulfillment of the Requirements in

Research II

(Special Science Program for Grade 8)

Mark Billy M. Cacal

Research Adviser

S.Y. 2018-2019
 CHAPTER I

Introduction

Background of the study

Medicinal plants are gifts from our environment that we need to

treasure. They play a vital role in human health. They are primitive medicines

during ancient times and up to present. They possess various natural constituents

that are effective in treatment of various diseases and preserve life.

A medicinal plant is any plant which, in one or more of its organ,

contains substances that can be used for therapeutic purposes, or which are

precursors for chemo-pharmaceutical semi-synthesis. When a plant is designated

as ‘medicinal’, it is implied that the said plant is a useful as drug therapeutic agent

or an active ingredient of a medicinal preparation. Medicinal plants may therefore

be defined as group of plants that possess some special properties or virtues that

qualify them as articles of drugs and therapeutic agents, and are used for medicinal

purposes.

Unfortunately, synthetic or commercialized medicines are widely

distributed and sometimes we ignore to use herbal plants. In urban places, most of

the people are engaged in using the synthetic medicines because there’s no process

to undergo before taking it wherein they do not know that these medicines has

adverse effects to human health. There are still people who appreciate the herbal

plants in their life. Those people who continue to utilize herbal plants are the people
living on rural places. They traditionally utilized medicinal plants in curing their

illnesses because of its effectiveness based on their own experiences and

knowledge. Old folks believe that herbal medicines are essential that synthetic

medicines because of its natural constituents where it prolongs their life span. It is

common to us that our ancestors have longer life because they took natural way in

taking their foods and processing their medicines.

There are many species of passion fruit (Passiflora sp.) which are

utilized as herbal plants. The passion fruit (Passiflora sp.) is a woody vine with

intricate blue, purple, red, or white flowers. There are several species which act as

sedatives and anxiolytics and are sometimes consumed as a tea or herbal

supplement. Some passion fruit (Passiflora sp.) plants species sometimes contain

small amounts of monoamine oxidase inhibitor (MAOI) chemicals, potentially

interacting with other chemicals and medications. This shows the contribution of

passion fruit (Passiflora sp.) to preserve the health of human through its natural

components. They are also the best source of man’s protection.

Human infections particularly those involving microorganisms that

is, bacteria, fungi, viruses, nematodes can cause serious infections in tropical and

subtropical countries of the world. In recent years, multiple drug resistance in

human pathogenic microorganisms has been developed due to indiscriminate use

of commercial anti-microbial drugs commonly used in the treatment of such

diseases. Plants have been the basis of many traditional medicines throughout the

world for thousands of years and have continued to provide new remedies to

mankind.
 Passion fruit (Passiflora sp.) are also used in culinary purposes in a way

that the fresh fruits are cut in half and the succulent pulp is scooped out with a spoon

for mixing with other fruits in making a salad. The extracted juice is a tasty drink

and the leaves of passion fruit are also consumed as a leafy vegetable.

Microbes are part of our lives they are present mostly in soil, water,

sewage and air. Our body consists of bacteria which are called pro bacteria where

in they live in our body without harming, they are called as the normal flora of the

body. But there are lots of microbes which bring diseases to human health and if

untreated may lead to death. Some are dangerous but some can be treated easily.

Statement of the Problem

This study will be conducted to determine the antibacterial activity of

passion fruit (Passiflora sp.) for eye irritations.

Specifically it aimed to answer the following:

1.Which is more effective between ethanolic and crude extracts of passion fruit

(Passiflora sp.) against Staphylococcus aureus, Streptococcus pneumonia and

Pseudomonas aeruginosa that causes eye irritations?

2.What is the first hand information about the passion fruit (Passiflora sp.)

according to the people in Antagan 1, Tumauini, Isabela.

Hypothesis

Passion fruit (Passiflora sp.) is effective for curing eye irritations.

This study aims to determine the antibacterial property of leaf extracts

passion fruit (Passiflora sp.) for eye irritations.

Specifically the study aimed to:

1.Test the efficacy and compare the antibacterial activity of passion fruit

(Passiflora sp.) extracts against Staphylococcus aureus, Streptococcus pneumonia

and Pseudomonas aeruginosa that causes eye irritations.

2.Determine the first hand information about the passion fruit (Passiflora sp.)

according to the people in Antagan 1, Tumauini, Isabela.

Significance of the Study

This study will be conduct to test the efficacy of passion fruit

(Passiflora sp.) as herbal plant in combating the different bacteria such as

Staphylococcus aureus, Streptococcus pneumoniae and Pseudomonas aeruginosa

which brings drastic effects on the sight of human. It also aims to provide

alternative medicine to the people who are engaged in synthetic drugs against eye

irritations.

Scope and Delimitations

The main focus of this study is on the determination of antibacterial

property of passion fruit (Passiflora sp.) ethanolic extraction of leaves through

Pseudomonas aeruginosa and the efficacy of the antibacterial property of leave

extract through different bacteria. It was also ocused on the determination of the

first hand information of the people about the passion fruit (Passiflora sp.)

Definition of Terms

Passiflora edulis. Is a vine species of passion flower that is native to southern

Brazil through Paraguay and northern Argentina.It is cultivated commercially in

tropical and subtropical areas for its sweet, seedy fruit, commonly called passion

fruit.

Extract. Is a substance made by extracting a part of a raw material, often by using

a solvent such as ethanol or water. Extracts may be sold as tinctures, absolutes or

in powder form.

Bacteria. A member of a large group of unicellular microorganisms which have

cell walls but lack organelles and an organized nucleus, including some which can

cause disease.
 CHAPTER II

REVIEW RELATED LITERATURE

Origin and Distribution

Traditionally and up to present passion fruit (Passiflora sp.) are

utilized by the people as food and medicine where they occur. The genus Passiflora

belongs to Passifloraceae family includes the passion fruit (Passiflora sp.) is the

largest and the most widespread genus of tropical flora. About 400 species within

this genus are grouped into 21 subgenera. More than 350 species have been found

in tropical regions and rain forests of South America and 60 of them are edible

species. Passion fruit (Passiflora sp.) is an important fruit crop in many tropical and

subtropical countries due to its edible fruits, ornamental use and medicinal

properties. The genus also contains some species of ornamental use and medicinal

properties as sedatives, antispasmodics and antibacterial.

The genus Passiflora consists of 500 species that are mostly found in

warm and tropical regions. Passiflora comes from Latin word “Passio” that was

first time discovered by Spanish discoverers in 1529 and was described as a symbol

for “Passion of Christ”.

The origin of the passion fruit (Passiflora sp.) is South America. It has

been observed in the coastal mountain rages of Panama, Brazil. It is native to

northern South America and West Indies. Widespread throughout the tropics and
serious in Southeast Asia; furthermore, a weed in the Pacific Region, West Africa

and Central America and introduced to Java a long time ago.

Morphological Characteristics

According to Patel et al., (2009), passion fruit (Passiflora sp.) is

herbaceous perennial vine with distinctive three-lobed leaves attractive flowers that

may reach 6 ½ feet or more in length. Passion fruit (Passiflora sp.) is primarily

found along the edges of woods and fields, but is coming noticeable in many of

Virginia’s agronomic crops, especially conservation tillage is practiced. The roots

initially develop as a taproot but eventually develop a very deep perennial rootstock

from which sprouts can emerge. Its leaves are arranged alternately along the stem.

Individual leaves are divided into three (only sometimes five) lobes that arise from

a common point (palmately lobed). Leaves are approximately 2 ½ to 5 ½ inches

long and wide and occur on petioles. The stems can trail along the ground or climb

on other vegetation. Solitary flowers arise from the area between and leaf petioles.

Flowers range from 2 to 4 inches in length and are very attractive and light purple

to lavender in colour. The nearly round or ovoid fruit, 1-1/2 to 3 inches wide, has a

tough rind that is smooth and waxy and ranging in hue from dark purple with faint,

fine white specks, to light yellow or pumpkin-color. Within is a cavity more or less

filled with an aromatic mass of double walled, membranous sacs containing orange-

colored, pulpy juice and as many as 250 small, hard, dark brown or black, pitted

seeds.

Uses
 According to Waltz (2004), passion fruit (Passiflora sp.) was an official

herb of the United States of America. Passion fruit is used to treat hyperactivity,

pre menstrual syndrome, Parkinson’s disease, nervous tension and the infusion is

used to treat eye infections and eye strain.

Passion fruit (Passiflora sp.) apparently exerts beneficial effects on

the nervous system and nutrition to the nervous. Its cooling properties help relieve

symptoms related to excess heat in the system. Its relaxing effects in the chest

relieve spasm and soothe irritating and nervous coughs. Passion fruit (Passiflora

sp.)appears to be generally free of side effects when taken alone at the

recommended dose.

Medicinal usage of passion fruit (Passiflora sp.) has been reported

in the traditional systems of medicine such as Ayurveda and has been used

extensively for treatment of some diseases like as anxiety, insomnia, convulsion,

sexual dysfunction, cough and cancer.

As cited by Ingale et al., (2010), the leaf extracts of passion fruit

(Passiflora sp.) exhibit better antibacterial activity than the fruits (Afolayan and

Meyer, 1997). Meyer et al., (1991), earlier reports focused on the antibacterial

activity of passiflora which has got activity against the Pseudomonas tetrandra,

Escherichia coli, Bacillus subtillis and Pseudomonas aeruginosa.

Tribal use of passion fruit (Passiflora sp.) showed the

effectiveness of the plant in many countries in Europe, U.S, and Canada, they use

passion plant leaves to tranquilize and settle edgy nerves has been documented for
over 200 years. It was also employed for colic, diarrhea, dysentery, insomnia,

neuralgia, eye disorders, epilepsy and convulsions, and muscle spasms and pain

and has antibiotic activity. Who would teach that passion fruit (Passiflora sp.) is

effective against those diseases that can cause drastic effects on human health?

Most of the species of passion fruit (Passiflora sp.) are edible; many

of them are also consumed as tea. According to Rosario (2005), the fruit of

Passiflora are used in the presentation of beverages or fruit juices. This shows the

relevance of the plant in the production of drinks.

According to the people of Antagan I, Tumauini, Isabela, they used

the leaf extracts as alternative remedy in combating eye irritations which they say

that this plant is very effective in treating their eyes. They employed the plant

extract in red eyes, blared of sight, and for sore eyes. According to them the leaves

extract cleans their eyes and there is no adverse effect to them. Many people in the

area have different ways on how they apply the extract. They used clean clothes to

gather the extract and they directly employ on the eyes that is irritated. Some say

that they used this once before going to bed and it depends upon the severity of the

irritations.

Based on the most current research, passion fruit (Passiflora sp.)

plant contains a variety of compounds, including alkaloids, phenols, glycoside,

flavonoids, and cyanogenic constituents which possess various medicinal

properties that are utilized by many people.

According to Sangvik (2012), the microbes that are found in our

bodies are known as the indigenous or the normal flora of the body. They consist

of bacteria, archaea, viruses, fungi and protists. During our birth, a life-long

symbiosis between and the microbes at a specific body site, including age, gender,

diet, hygiene, clothing, climate, and occupation and living conditions.

Our eyes have a vital role in our daily lives. An important organs that

help us to survive to this world in which there are vast number of microorganisms

such as bacteria, viruses and fungi which could affect the sight of individual and

may bring drastic effects or lost of sight if it is contracted with our eyes and if left

untreated.

Redness can be caused by inflammation, infection or irritation if the

conjunctival membranes that line the prosterior surface of the lids and the anterior

of sclera. In primary care practice, one of the most causes is conjunctivitis, which

can result from infection.

Red eye is one of the most common ophthalmologic conditions in the

primary care setting. Bacterial conjunctivitis is highly contagious and is most

commonly spread through direct contact with contaminated fingers.

Staphylococcus aureus infection often causes acute bacterial conjunctivitis in

adults, whereas Streptococcus pnuemoniae infections are common in children (H.

Cronau, MD., et al., 2010). Conjunctivitis is an inflammation or redness of the

can be caused by infection, allergic reactions or physical agents. Conjunctivitis is a

common eye problem because the conjunctivae are continually exposed to

microorganisms and environmental agents that can cause infections or allergic

reactions. Conjunctivitis can be acute or chronic depending upon how the condition

lasts, the severity of symptoms, and the type of organisms or agent involved. Other

names for conjunctivitis include pink eye and red eye.

Bacterial conjunctivitis is caused by a wide range of gram-positive

and gram-negative organisms, but the former predominate. Acute bacterial

conjunctivitis typically has an abrupt onset, develops in one eye initially but spreads

to the opposite eye within 48 hours, and is manifested as tearing and ocular irritation

at the outset. A mucopurulent or prulent discharge develops within one or two days,

with a collection of debris at the base of the lashes and matting of the lids,

particularly on awakening.

Staphylococcus aureus

According to Siegrist (2001), the name staphylococcus aureus

comes from the Greek words “ staphyle “, meaning a bunch of grapes, “coccus”,

which means round-shape, and “aureus”, for golden, because most colonies have a

characteristic orange-yellow coloring on the traditionally-used agar plates.

Staphylococci may be airbone and can occur in both animals and humans, in

sewage, water, milk, or food, and on environmental surfaces or food equipment.

They also grow in temperature range of 15-45 degree Celsius and in a medium with

up to 15% sodium chloride. Staphylococcus aureus is frequently a part of our skin

(1964), Staphylococci can produce disease both through their ability to multiply

and spread widely in tissues and through their production of many extracellular

substances.

Staphylococci are easily spread between animals and under certain

conditions to humans as well as by skin to skin contract, but also by contract of

excretions which contain staphylococci, such as saliva or aerosols released during

sneezing and coughing. Moreover, staphylococci may spread by animal products,

such as non-pasteurized milk.

Streptococcus pneumoniae

Streptococcus pneumoniae is a diverse species that is classified into

immunologically distinct serotypes based on capsular polysaccharides present on

the bacterial surface. Over 90 different serotypes have been identified.

Some are related to each other and belong to single serogroup such as serotypes

!5B and 15C within serogroup 15, whereas some serogroups consist of a single

serotype 3. S. pneumoniae is typically grown on blood agar. Pneumonia is the

leading killer of children under the age of five worldwide, and the gram-positive

bacterium S. pneumoniae (the pneumococcus) is the most common aeotilogical

agent. S. pneumoniae is frequently carried in the nasopharynx of children and

though colonization is asymptomatic. Furthermore, carriage serves as a

reservoir for maintaining strains of S. pneumonia in human population.

Eye infections by S. pnuemoniae include corneal ulcer, dacryocystitis and

serotypes are associated with majority of these infections. Moreover, up to 65

percent of normal population carry S. pnuemoniae is one of the predominant

bacterial pathogens responsible for eye infections in different parts of the world

including India. The conjunctival sac and lid margins of the eye harbor a variety of

bacteria as normal flora.

The bacterium S. pnuemoniae is a major cause of bacterial

endophthalmitis, especially after ocular surgery. Intravittreal vancomycin or

ceftazidime is often administered to treat pneumococcal endophthalmitis .

However, despite treatment, this disease is particularly devastating to the eye and

can result in evisceration and poor visual outcome. Little known about the factors’

responsible for the pathogenesis of the pneumoccal endophthalmitis, except that the

S. pneumonia toxin pneumolysin is responsible for some of the retinal damage

caused by the bacterium. The polysaccharide capsule surrounding of Streptococcus

pneumoniae has been well established as a major virulence factor in non-ocular

pneumococcal infections such as systematic infections, lung infections and

bacteremia. The function of the capsule in these infections is to allow the bacterium

to evade phagocytosis.

According to Burton et al., (2011), the infected humans are the

reservoirs of S. pneumoniae. Human to human transmission occurs via contract

with eye and respiratory charges, contaminated fingers, facial tissues, clothing, eye

makeup, eye medications and lens wetting and lens cleaning agents.
 Streptococci are a spherical microorganisms, characteristically

arranged in chains and widely distributed in nature. Some are members of normal

human flora; others are associated with important human diseases attributable in

part to infection by streptococci.

Pseudomonas aeruginosa

Aside from Staphylococcus aureus and Streptococcus pneumoniae

which causes eye irritations, Pseudomonas aeruginosa is also a microorganism

which brings irrritations to sight of humans. Due to these bacteria it may damage

the cornea, often permanently. Enzyme produce by the bacteria can rapidly destroy

the eye. Infections may result from contamination of contract lenses or contract lens

solutions.

Pseudomonas group is composed of gram-negative bacteria. They

occur widely in soil, water, sewage and air. Pseudomonas aeruginosa is frequently

present in small numbers in the normal intestinal flora. It is also found in the human

skin. This organism is pathogen only when introduced into areas devoid of normal

defenses or when participating in mixed infections. Aside from the infection of the

eye which leads to rapid destruction of the bulbus, occurs most commonly after

injury or surgical procedures, Pseudomonas aeruginosa also present in the

wounds, giving rise to blue-green pus, meningitis, when introduced by catheters

and instruments or in irrigating solutions (E. Jawetz 1964). According to the study

of Mesaros et al., (2007), the prevalence of colonization rates can be encountered

antimicrobial agents. Colonization is common in the respiratory tract of

mechanically ventilated patients, in the gastrointestinal tract of patients receiving

anticancer chemotherapy, and on the skin of burn patients. They are also present in

sinks, mops, disinfectant solutions, respiratory equipments, food mixers and other

moist environment can act as reservoir of P.aeruginosa in the hospital setting. In

USA,, P. aeruginosa was the second most common cause of nosocomial

pneumonia, the third most common nosocomial urinary tract infections, and the

seventh most common cause of nosocomial bacteraemia. In Europe, P, aeruginosa

was found to be the third most common isolate from nosocomial infections in

intensive care units (ICUs).

Spencer (1953) believed that Pseudomonas aeruginosa is an

“opportunist” having no invasive power of its own, being controlled by the defenses

of the body. The devastating effects of the eye by Pseudomonas aeruginosa have

received much comment in opthalmological literature. The most common infection

of these bacteria is corneal ulcer, which spreads with frightening rapidity to

panophthalmitis. The infection often follows penetration of the cornea by a foreign

body which becomes embedded there. When introduce to the cornea, however, as

in penetration by a foreign body or in contaminated medicines, it acts extreme

virulence, in many cases causing blindness.

According to the recent study of Ochoa in 2013, Pseudomonas

aeruginosa can survive with low levels of nutrients and grow in temperatures
ranging from four to forty two degrees Celsius. These characteristic allow it to

attach itself and survive on medical equipment and on other hospital surfaces,

which favors the beginning of infections in immunocompromised patients.

Infections due to Pseudomonas aeruginosa are difficult to eradicate because of their

elevated intrinsic resistance as well as their capacity to acquire resistance to

different antibiotics.
 CHAPTER III

Research design

Our study titled “Antibacterial Activity of Passion fruit (Passiflora

sp.) Leaf Extracts” will be using Single Group Design because we will use one leaf

extract with three sample replications.

The aim of this study is to determine the antibacterial activity of leaf

extract of Passion fruit (Passiflora sp.) for eye irritations.

Research environment

The Passion fruit (Passiflora sp.) will be collected at Antagan 1,

Tumauini, Isabela and the entire process will be conducted in the Botany Division

of National Museum for identification and authentication.

Materials and Equipment

The Passion fruit (Passiflora sp.) is the main materials that we need

for the study. The leaves of the plant were washed and pounded with mortar and

pestle to collect the extracts. The antibacterial activity of the leave extracts was

measured based on their growth of inhibition. Using vernier caliper to measure the

standard measurement of zone of inhibition.

Procedure

1.Collect the Passion fruit (Passiflora sp.)

2.Use the mortar and pestle to collect the extracts.

4.Placed it in the bottle.

5.Soaked 92g of plant leaves in 80% ethanol.

6.After 24 hours of soaking, the extracts were collected and subjected to

evaporation to remove the water.

Flowchart

Gathering of

materials

Preparation of leaf

extract

Conducting the

activity
 Antibacterial

activity of Passion

fruit (Passiflora sp.)

leaf extracts

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