TITLE

Tridax Daisy (Tridax procumbens) Leaf Extract as an Alternative Antibacterial

Agent against Klebsiella pneumoniae

RATIONALE

Klebsiella pneumoniae is an emerging pathogen associated with significant

mortality nowadays (Wilmington, 2009). K. pneumoniae have characteristics of clinical

manifestations. It is a superbug that causes a scope of diseases, contingent upon which

body part it infects. The researchers’ findings highlight the urgent need to develop low

cost and efficient strategies for prevention and infection control of the bacteria.

The researchers had the willingness and motivation in this study because

Klebsiella pneumoniae can progress into severe bacterial infections leading to different

diseases and disorders. Fewer and fewer treatments are available for infections by K.

pneumoniae because of the developing resistance mechanisms. These diseases have

been a great problem or widespread particularly in the Philippines; so this way, the

researchers would contribute in the health provision in Filipinos.

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STATEMENT OF THE PROBLEM

The study “Tridax Daisy (Tridax procumbens) Extract as an Alternative

Antibacterial Agent against Klebsiella pneumoniae” generally aims to produce an

alternative antibacterial agent against Klebsiella pneumoniae.

The study specifically seeks:

1. To identify the active chemical components present in Tridax daisy

a. flavonoids,

b. alkaloids, and

c. tannins

2. To determine the effectiveness of the variation of tridax daisy to the normal

saline solution in terms of zone of inhibition.

3. To test the significant difference of the variation of tridax daisy to the normal

saline solution in terms of zone of inhibition.

Hypotheses

H0: There is no significant difference between the effectiveness of tridax daisy

to the normal saline solution in terms of zone of inhibition

H0: There is no significant difference between the variation of tridax daisy to

the normal saline solution in terms of zone of inhibition

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 RELATED LITERATURE AND STUDIES

Tridax Daisy (Tridax procumbens)

Tridax daisy is in the family of Asteraceae. The plant is annual or biennial

somewhat patently hispid herbs. The stem branched, creeping at base, sub erect or

trailing above. The leaves of the plants ovate-lanceolate, or elliptic-rhomboid, with a

cuneate base, obtuse or subacute, coarsely serrate or lobed, patently hispid, 2.5-7 cm

long. Marginal flowers 5-6 with pale yellow, 0.3 cm long ligules; disc flowers bright

yellow. The juice from the leaves of the tridax daisy possesses antiseptic, insecticidal

and parasitical properties. The crushed leaves are applied to arrest bleeding in bruises

and cuts. Leaves are also used for the treatment of bronchial catarrh, dysentery, and

diarrhea and for the restoration of hairs (Uddin, 2014).

Plants like tridax daisy have limitless ability to synthesize a vast array of

bioactive compounds which possess some bioefficacy (Jain, 2015). Plants are known to

provide biologically active molecules with reduced or no toxicity. Tridax procumbens is

employed as indigenous medicine for a variety of ailments (Khale & Dake, 2013). It has

been extensively used in Indian traditional medicine for wound healing, as

anticoagulant, antifungal and insect repellent, in diarrhea and dysentery (Ahmad et al.,

2016).

Aerial parts of the plant contain sterols, campesterol, stimasterol, β-sitosterol and

saturated and unsaturated C12-C22 fatty acids. Flowers contain flavonoids, luteolin,

glucoluteolin and quercetin (Ghani, 2003).

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 The extracts of tridax daisy have been reported to have various pharmacological

effects like mosquito repellant activity, leishmanicidal, hepatoprotective effect on liver

antioxidant system, immunomodulatory effect, wound healing activity and antiprotozoal

effects.

There are number of reports on different part of the plant and whole plant of

tridax daisy has been reported for its antimicrobial activity on various species of

bacterial. Antibacterial activity is mostly investigated by disc diffusion and agar-well

diffusion method (Krishnavignesh et al., 2012).

Klebsiella pneumoniae

Klebsiella pneumoniae is a type of Gram-negative bacteria that can cause

different types of healthcare-associated infections, including pneumonia, bloodstream

infections, wound or surgical site infections, meningitis, urinary tract infections (UTI),

nosocomial infection, rhinoscleroma and ozena, and chronic genital ulcerative disease.

K. pneumoniae is a superbug that causes a range of diseases, depending on which part

of the body it infects. Klebsiella pneumoniae is a bacterium that normally lives inside

human intestines where it does not cause disease. K. pneumoniae can cause a range

of different illnesses if it gets into other areas of the body. K. pneumoniae infections are

typically a nosocomial infection which means they are contracted in a hospital or

healthcare setting. Klebsiella infections is spread through exposure to the bacteria via

respiratory tract. Klebsiella infections are most well-known in hospitals spread through

person-to-person contact by contaminated hands of surrounded people in the hospitals,

whether it be an employee or a patient. Klebsiella is spread very easily and rapidly, but

not through the air. Healthcare settings are most vulnerable to Klebsiella infections due

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to the nature of procedures that allow easy access of bacteria into the body. People

who have weakened immune systems, or sick or injured people who are undergoing

procedures for various health issues, are more likely to get a Klebsiella infection.

Healthy people usually do not have to worry about getting K. pneumoniae infections,

according to the Centers for Disease Control and Prevention. People cannot contract a

K. pneumoniae infection by breathing the same air as an infected person because the

bacteria is not airbourne. K. pneumoniae is spread through direct person-to-person

contact, such as when someone with contaminated hands touches a wound. Infections

can also occur through the use of contaminated medical equipment. Patients in

healthcare settings also may be exposed to Klebsiella when they are on ventilators, or

have intravenous catheters or wounds caused by injury or surgery. Unfortunately, these

medical tools and conditions may allow K. pneumoniae to enter the body and cause

infection. Taking antibiotics over a long course of time can also increase a person’s risk

of getting Klebsiella infection (Castro, 2015).

Humans are the primary reservoir for K. pneumoniae. Carrier rates of K.

pneumoniae in the community range from five (5) to 38 percent in stool samples and 1

to 6 percent in the nasopharynx; Klebsiella species are rarely carried on the skin. Higher

rates of nasopharyngeal carriage have been noted in ambulatory alcoholic patients (Yu

& Chuang, 2016). The mortality rate may be as high as 50% in the case of pneumonia

caused by K. pneumoiae. The prognosis is worse for patients with alcoholism and

bacteremia. A patient administered a course of broad-spectrum antibiotic treatment is at

an even high risk due to the disruption of the normal flora of the bacteria in the body,

deeming it more susceptible to pathogens.

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 When dealing with hospital-acquired bacterial infections caused by K.

pneumoniae can arise in different parts of the body and in different forms of illness

depending on transmission. K. pneumoniae is responsible for 6-17% of UTI’s, 7-14% of

pneumonia, 4-15% of septicemia, 2-4% of wound infections, 4-17 nosocomial infections

in intensive care units, and 3-20% of all neonatal septicemia cases. All of these cases

rank within at least the top 11 in comparison to all other bacterial pathogens. In the

United States, people who suffer from alcoholism make up 66% of people affected by

community-acquired pneumonia. K. pneumoniae is now among the top eight (8)

pathogens in hospitals and is a rising issue among hospitals all around the world due to

antibiotc resistance. In humans, K. pneumoniae resides in the nasopharynx and in the

intestinal tract. Since gram-negative bacteria do not have good growth on human skin,

they are rarely found there in comparison to internal parts of the body. Carriers rated in

hospitalized patients were 19% in the pharynx, 77% in the stool, and 42% on the hands.

Even hospital employees had elevated rates of carriage to K. pneumoniae. These

findings were linked to the over usage of broad-spectrum antibiotics rather than delivery

of care. In 2011, an investivation of Klebsiella pneumoniae carbapenemase (KPC)-

producing Enterobacteriaceae was conducted in hospitals among patient with short and

long-term stays. Over a 1-year period, KPC-producing Enterobacteriaceae was found

throughout 4 counties in Indiana and Illinois. The source of the problem was found to be

within long-term facilities and patients. KPC has been found in a total of 44 states thus

far 14% of bacteremia cases are because by K. pneumoniae, which places it in second

place next to Escherichia coli for origins of gram-negative sepsis. Outbreaks of neonatal

septicemia and K. pneumoniae can be found worldwide. In Israel, a number of hospital

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facilities reported increases in KPC-producing Enterobacteriaceae beginning in 2006,

while the first case in the United States was reported in 2001 (Kappel, 2016).

Alkaloids

Alkaloids are a large and structurally diverse group of natural products of

microbial, plant and animal origin. Alkaloids are responsible for the beneficial effects of

traditional medicines such as cinchona bark, but also the harmful effects of poisons.

Alkaloids have inspired the development of several antibacterial drugs, with synthesis of

quinine serendipitously yielding the quinolones, structural alteration of azomycin yielding

metronidazole, and work with the quinolone scaffold yielding bedaquiline. Alkaloids are

present in other drugs as scaffold substructures. Alkaloids remain the focus of much

research, their development as antibacterial drugs pursued within academia, industry

and joint ventures. Alkaloids are found in bacteria, fungi, plants and animals, although

their distribution within each kingdom is quite limited. Alkaloids occur in ca. 300 plant

families, specific compounds typically confined to certain families. Alkaloids can occur

in any part of the plant, though specific compounds may be limited to a certain part.

More than 18,000 alkaloids have been discovered. Multiple roles have been attributed

to alkaloids in the above organisms, most related to self-preservation, inhibition of

competitors, or communication. In micro-organisms, for example, alkaloids act as

feeding deterrents, allelochemicals, autoinducers and siderophores. The inhibitory

effects in plants of alkaloids on glycosidase and trehalose metabolism deter herbivores,

and the ability to quench singlet oxygen confers protection against this toxic

photosynthetic byproduct. Alkaloids also act as phytoanticipins and phytoalexins,

protecting plants against infection. The great structural diversity, the presence of a basic

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nitrogen atom being the only unifying feature characterizes the alkaloids. Most alkaloids

possess just one (1) nitrogen atom, but some have up to five (5). Pharmacological

properties include analgesic, central nervous stimulant, central nervous depressant,

antihypotensive, antihypertensive, antipyretic, anticholinergic, antiemetic, oxytocic and

vasoconstrictor, antitumor and antimalarial activities. These activities are exploited both

in traditional medicine and modern medicine. Other alkaloids have been incorporated

into human culture as recreational drugs and drugs of abuse (Cushnie & Lamb, 2014).

The effect alkaloid from various plants or bacterial growth and efflux pump activity was

evaluated on some different strains of bacteria and it inhibited bacterial growth with

comparable effects to ampicillin, a standard antibiotic. Some alkaloids may serve as

potential courses of compounds that can act as lead compounds for the development of

plant based antibacterial and their adjunct compound (Mabhiza, Chitemerere, &

Makunganyama, 2016).

With reports of pandrug-resistant bacteria causing untreatable infections, the

need for new antibacterial therapies is more pressing than ever. Alkaloids are a large

and structurally diverse group of compounds that have served as scaffolds for important

antibacterial drugs.

Flavonoids

Flavonoids are ubiquitous in photosynthesizing cells and are commonly found in

fruit, vegetables, nuts, seeds, stems, flowers, tea, wine, and honey. For centuries,

preparations containing these compounds as the principal physiologically active

constituents have been used to treat human diseases. Increasingly, this class of natural

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products is becoming the subject of anti-infective research, and many groups have

isolated and identified the structures of flavonoids possessing antifungal, antiviral and

antibacterial activity. Moreover, several groups have demonstrated synergy between

active flavonoids as well as between flavonoids and existing chemotherapeutics.

Reports of activity in the field of antibacterial flavonoid research are widely conflicting.

However, several high-quality investigations have examined the relationship between

flavonoid structure and antibacterial activity and these are in close agreement. In

addition, numerous research groups have sought to elucidate the antibacterial

mechanisms of action of selected flavonoids. The activity of quercetin, for example, has

been at least partially attributed to inhibition of Deoxyribonucleic acid (DNA) gyrase. It

has also been proposed that sophoraflavone G and (−)-epigallocatechin gallate inhibit

cytoplasmic membrane function, and that licochalcones A and C inhibit energy

metabolism. Other flavonoids whose mechanisms of action have been investigated

include robinetin, myricetin, apigenin, rutin, galangin, two (2), four (4), two (2) -

trihydroxy- five (5) -methylchalcone and lonchocarpol A. These compounds represent

novel leads, and future studies may allow the development of a pharmacologically

acceptable antimicrobial agent or class of agents. Flavonoids are ubiquitous in

photosynthesising cells and therefore occur widely in the plant kingdom. Flavonoids are

found in fruit, vegetables, nuts, seeds, stems and flowers as well as tea, wine, propolis

and honey, and represent a common constituent of the human diet. In the US, the daily

dietary intake of mixed flavonoids is estimated to be in the range 500–1000 mg, but this

figure can be as high as several grams for supplementing diets of the humankind with

flavonoids or flavonoid-containing herbal preparations. The function of flavonoids in

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flowers is to provide colors attractive to plant pollinators. In leaves, these compounds

are increasingly believed to promote physiological survival of the plant, protecting it

from, for example, fungal pathogens and UV-B radiation. In addition, flavonoids are

involved in photosensitization, energy transfer, the actions of plant growth hormones

and growth regulators, control of respiration and photosynthesis, morphogenesis and

sex determination.

The antibacterial activity of flavonoids is being increasingly documented. Crude

extract from plants with a history of use in folklore medicine have been screened in vitro

for antibacterial activity by many research groups. Many phytochemical preparations

with high flavonoid content have also been reported to exhibit antibacterial activity.

Small number of groups have investigated the relationship between flavonoid structure

and antibacterial activity have been able to identify common structural features among

active compound. Individual antibacterial flavonoids have multiple cellular targets, rather

than one specific site of action. These common structural features may be simply

necessary for flavonoids to gain proximity to or update into the bacterial cell (Cushnie &

Lamb, 2005).

Tannins

Tannins are astringent, pitter plants polyphenols that either bind and precipitate

or shrink proteins. The term tannin refers to the use of tannin in tanning animal halides

into leather; however the term is widely applied to any large polyphenolic compound

containing sufficient hydroxyls and other suitable groups to form strong complexes with

proteins and other macromolecule. Many biological activities and antibacterial

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promoting effect have been reported for plant tannin and flavonoids and now

investigated increasingly. Tannins have traditionally been considered anti-nutritional but

it is now known that their beneficial or anti-nutritional properties depend upon their

chemical structure and dosage. The new technologies used to analyze molecular and

chemical structures have shown that a division into condensed and hydrolysable

tannins is far too simplistic (Al-Ani & Mohammed, 2008).

Tannins are water-soluble polyphenolic compounds of variable molecular weights

abundantly found in nature which have the ability to precipitate proteins (Spencer et al.,

1988; Cowan, 1999). Many studies of phenolic compounds (resveratrol, quercetin, rutin,

catechin, proanthocyanidins) have been present in the last few years, most of these

works were directed to improvements of human health and they demonstrate that

tannins have multiple biological activities, including cardioprotective, anti-inflammatory,

anti-carcinogenic, antiviral, and antibacterial properties attributed mainly to their

antioxidant and antiradical activity (Frankel et al., 1993; Teissedre et al., 1996; Santos-

Buelga and Scalbert, 2000). Tannins are also able to reduce the risk of livestock

disease and transmission of zoonotic pathogens in a sustainable and environmentally

friendly manner. Recent reports of the use of tannin in poultry show promising results

(Van Parys et al., 2010; Anderson et al., 2012; Redondo et al., 2013b; Tosi et al., 2013).

Tannins have shown potential antiviral (Lin et al, 2004), antibacterial (Akiyama et al.,

2001; Funatogawa et al., 2004), and antiparasitic effects (Bhagavathi et al, 1999; Yang

et al, 2000; Tanimura et al, 2005). There are several reports on the use of tannins in

treating various ailments in humans, including diarrhoea, gastric ulcers, snake bites and

wounds (e.g. Perera et al. 2001).

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 Tannin toxicity for fungi, bacteria and yeasts is reviewed and compared to toxicity

of related lower molecular weight phenols. The dependence of toxicity is examined on

tannin structure. The different mechanisms proposed so far to explain tannin

antimicrobial activity include inhibition of extracellular microbial enzymes, deprivation of

the substrates required for microbial growth or direct action on microbial metabolism

through inhibition of oxidative phosphorylation. A further mechanism involving iron

deprivation is proposed. Many microorganisms can overcome plant defences based on

tannins. These microorganisms may detoxify tannins through synthesis of tannin-

complexing polymers, oxidation, tannin biodegradation or synthesis of siderophores

(Scalbert, 1991).

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 MATERIALS AND METHOD

The study “Tridax Daisy (Tridax procumbens) Leaf Extract as an Alternative

Antibacterial Agent against Klebsiella pneumoniae” generally aims to produce an

alternative antibacterial agent against Klebsiella pneumoniae. The tridax daisy leaves

will be coming from the field of Quezon Science High School. The researchers will

gather the materials needed to be used for the extraction which are the containers,

strainer, filter paper, and the mortar and pestle.

The researchers will gather one (1) kg of tridax daisy. The leaves of Tridax daisy

will be separated and will be washed with tap water. The researchers will leave the

leaves of tridax daisy for some time until it is dry. The plant will be shredded in a

container and be added with 50 ml of methanol. The researchers will pound it with the

use of mortar and pestle and then squeeze it into an empty container with the strainer to

obtain the extract from the leaves. The researchers will leave the container exposed to

heat for the methanol to evaporate and to get pure extract of the tridax daisy leaves.

The extract will be introduced to the experimental units through disc diffusion

sensitivity testing and tube dilution testing that will occur at the Calayan Educational

Foundation Incorporated (CEFI) with the guidance of a medical technologist.

Treatments and General Procedure

The researchers will have five (5) treatments. Treatment A will have 100% tridax

daisy leaf extract, treatment B will have 75% tridax daisy leaf extract and 25% normal

saline solution, treatment C will have 50% tridax daisy leaf extract and 50% normal

13
saline solution, treatment D will have 25% tridax daisy leaf extract and 75% normal

saline solution, while treatment E will have 100% normal saline solution.

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 ABSTRACT

Nowadays, bacteria are mostly the main cause of the diseases the people are

starting to get. The humans are experiencing different infections and diseases such as

lung diseases, blood infections, urinary tract infections, wound infections, meningitis and

many more due to the bacteria called Klebsiella pneumonia.

The researchers want to provide a medicine with a low cost and can be seen

easily that can help avoid diseases connected with the bacteria K. pneumoniae. The

researchers are to find the chemical components found in Tridax daisy (Tridax

procumbens) that can help kill the bacteria K. pneumoniae. The researchers will find the

effectiveness of Tridax daisy (T. procumbens) as an alternative antibacterial agent

against K. pneumoniae in terms of zone of inhibition. The researchers are to use Tridax

daisy from the field of Quezon Science High School.

The researchers will gather the materials and the tools needed for the research.

The researchers will make a methanolic extract of Tridax daisy (T. procumbens). The

researchers will have five (5) treatments, of which four (4) are experimental treatments

and one (1) is a control treatment. Treatments A, B, C and D will have 25%, 50%, 75%,

100% extract of Tridax daisy, respectively, while treatment E with normal saline

solution.

The results of the phytochemical analysis are: Flavonoids are the most abundant.

Alkaloids, Sterols, Triterpenes are moderately abundant. Saponins, Tannins, and

Glycosides are traced constituents.

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Background of the Study

Infectious diseases remain a major cause of death, disability, and social and

economic disorder for millions of people throughout the world (Nizet &Esko, 2009).

Bacteria or viruses are usually the cause of many human infections. Bacteria are tiny

single-celled organisms. It can be beneficial but some are responsible for a range of

infections (Better Health Channel, 2014). The spectrum of clinical syndromes includes

lung diseases, bloodstream infections, wound infections, urinary tract infections and

meningitis (Genetic and Rare Diseases Information Center, 2014). Gram-negative

bacterium is one of the main causes of infections. There is an increase in the likelihood

of nosocomial infection with Klebsiella species (Qureshi, 2015). One of the most

common species of bacteria that cause problems in healthcare today is Klebsiella

pneumonia.

Klebsiella pneumoniae is a major human pathogen that has been implicated in

infections in healthcare settings over the past few decades. Antimicrobial treatment of

K. pneumoniae infections has become increasingly difficult as a consequence of the

emergence and spread of strains that are resistant to multiple antimicrobials

(Moradigaravand et al., 2016). Recently, invasive infections due to K. pneumoniae were

geographically confined to Southeast Asia and South Africa. Nowadays, it has been

recorded that Klebsiella pneumoniae is the main cause of severe infections and is

associated with a high mortality rate which is around 50%.

Klebsiella pneumoniae is a type of Gram-negative bacteria that can cause

different types of healthcare-associated infections (Jackson, 2006). It was found out

16
alkaloids, flavonoids, and tannins are considered to be the most important bioactive

compounds of plants for their potential antimicrobial trait (Dahiya &Purkayastha, 2012).

The phytochemical screening revealed that one of the plants that have the presence of

these constituents is Tridax Daisy (Tridax procumbens). Tridax daisy (T. procumbens) is

a common medicinal herb used by ethno-medical practitioners, belonging to a family

Asteraceae. It is best known as a widespread weed and pest plant. The plant is a

procumbent herb and is valued for its pharmaceutical properties (Sahoo & Chand,

1998). It is a common weed that grows in open places, coarse textured soils of tropical

regions, sunny dry localities, fields, waste areas, meadows and dunes (Khan, 2008).

The said bacterium is an emerging pathogen associated with significant mortality

nowadays, as said (Wilmington, 2009). Most notoriously, it is a prominent nosocomial

pathogen that led to different infections, diseases and disorders such as urinary tract

infections, blood stream infections, wound or surgical site infections, meningitis,

rhinoscleroma, pyogenic liver abscess and pneumonia (Brisse, Fevre & Passet, 2009).

Community-acquired pneumonia (CAP) is one of the most common life-threatening

infections, with most deaths occurring in developing countries (Horby et al., 2014). In

the recent two decades, K. pneumoniae has become the most common causative

pathogen of pyogenic liver abscess in Asian countries. Here in the Philippines,

pneumonia, which is also caused by the said pathogen, is considered in the top 5

leading causes of death and infant mortality up to the present.

Because of this problem, the researchers’ findings highlight the urgent need to

develop strategies for prevention and infection control in a low cost and efficacy. The

researchers had the willingness and motivation in this study because the acquisition of

17
Klebsiella pneumoniae can progress into severe bacterial infections leading to different

diseases and disorders. These diseases have been a great problem or threat and

widespread particularly in the Philippines; so this way, they would contribute in the

health provision in Filipinos.

18
Statement of the Problem

The study “Tridax Daisy (Tridax procumbens) Extract as an Alternative

Antibacterial Agent against Klebsiella pneumoniae” generally aims to produce an

alternative antibacterial agent against Klebsiella pneumoniae.

The study specifically seeks:

1. To identify the active chemical components present in Tridax daisy

a. Flavonoids,

b. Alkaloids, and

c. Tannins

2. To determine the effectiveness of the variation of tridax daisy to the normal saline

solution in terms of zone of inhibition

Using Disc Diffusion Sensitivity Testing

3. To determine the effectiveness of the variation of tridax daisy to the normal saline

solution in terms of zone of inhibition

Using Tube Dilution Testing

4. To test the significant difference of the variation of tridax daisy to the normal

saline solution in terms of zone of inhibition

Using Disc Diffusion Sensitivity Testing

5. To test the significant difference of the variation of tridax daisy to the normal

saline solution

Using Tube Dilution Testing

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HYPOTHESES

H0: There is no significant difference between the effectiveness of tridax daisy to

the normal saline solution in terms of zone of inhibition using disc diffusion

sensitivity testing

H0: There is no significant difference between the effectiveness of tridax daisy to

the normal saline solution in terms of zone of inhibition using tube dilution testing

H0: There is no significant difference between the variation of tridax daisy to the

normal saline solution in terms of zone of inhibition using disc diffusion sensitivity

testing

H0: There is no significant difference between the variation of tridax daisy to the

normal saline solution in terms of zone of inhibition using tube dilution testing

Significance of the Study

Different types of infections have been increasing all over the world and so are

the needs of new antibacterial drugs. A lot of people are starting to suffer from these

infections. Different commercial antibiotic drugs are given to the people and what a lot

of people do not know that most of these drugs are not safe. Klebsiella pneumoniae is

one of the bacteria that can cause a range of different illnesses. Most people who have

been suffering from the illness brought this type of bacteria rely on the chemically made

antibiotics and most do not know how more efficient and safe natural alternatives are.

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 Tridax daisy (T. procumbens) is found all over the Philippines and can be easily

be found. Tridax daisy (T. procumbens) contains flavonoids, alkaloids and tannins that

can help eliminate bacteria.

Scope and Limitations

The study “Tridax Daisy (Tridax procumbens) Leaf Extract as an Alternative

Antibacterial Agent against Klebsiella pneumoniae” covers the testing of the

effectiveness and significant difference of tridax daisy (T. pocumbens) leaf extract

against Klebsiella pneumoniae in terms of zone of inhibition. The study has five (5)

treatments, of which four (4) are experimental treatments and one (1) is control

treatment. Each treatment consists of three (3) trials. Laboratory testing and observation

was conducted at Calayan Educational Foundation Incorporated (CEFI) in Lucena City

with the assistance of a licensed medical technologist.

The phytochemical analysis of tridax daisy (Tridax procumbens) leaves was

conducted at Department of Science and Technology (DOST), Bicutan, Taguig City.

The tridax daisy (T. procumbens) leaves used in the laboratory testing and

phytochemical analysis were gathered in the field of Quezon Science High School.

The plant verification of trdax daisy leaves was done at the Museum of Natural

History in the University of the Philippines Los Baños, Los Baños, Laguna.

The study only covers the effect of Tridax Daisy (Tridax procumbens) leaf extract

against Klebsiella pneumoniae. It will not cover the effects of tridax daisy against other

bacteria; it will also not be used as treatment for diseases and other body organs.

Additionally, the side effects of tridax daisy are not included in this study.

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