wpaper (1)
-
Upload
wendell-flores -
Category
Documents
-
view
222 -
download
0
Transcript of wpaper (1)
-
7/28/2019 wpaper (1)
1/22
ANTIBACTERIAL ACTIVITY OF GUYABANO (Annona muricata) LEAVES
EXTRACT ON Staphylococcus epidermidis
A Science Investigatory Project Proposal
Submitted by:
Wendell Flores
II-Daffodil
BICOL REGIONAL SCIENCE HIGH SCHOOL
TUBURAN, LIGAO CITY
IN PARTIAL FULFILLMENT OF THE REQUIREMENT IN RESEARCH I
Submitted to:
Mrs. Ester Llobet-Maligaya
S.Y 2012-2013
1 | P a g e
-
7/28/2019 wpaper (1)
2/22
TABLE OF CONTENTS
I. Introduction
a) Statement of the Problem
b) General Problem
c) Specific Problem
d) Hypothesis
Operational
Null
e) Scope And Limitations
f) Definition of Terms
g) Conceptual Framework
h) Endnotes
II. Review of Related Literatures
a) Annonaceous acetogenins
b) Guyabano
c) Antibacterial
d) Staphylococcus Epidermidis
e) Biofilms
2 | P a g e
-
7/28/2019 wpaper (1)
3/22
f) End Notes
III. Methodology
a) Methods of research
b) Methods of collecting data
c) Statistical Treatment
d) Materials
e) Procedure: Extraction
Concentration 1
Concentration 2
Concentration 3
Concentration 4
Concentration 5
Concentration 6
Concentration 7
3 | P a g e
-
7/28/2019 wpaper (1)
4/22
Introduction
BACKGROUND OF THE STUDY
Guyabano (Annona muricata) are slender trees that are 5 to 10 meters in height and 15
centimeters in diameter. The trunk of the guyabano are mostly straight , the barks are smooth
and the leaves are alternately 7.6-15.2 cm long and 2.5-7.6 cm wide .Guyabano is commonly
used as herbal medicines especially its leaves. It contains different minerals and vitamins like
vitamin C, B1, B2, phosphorous, potassium, fiber and iron. 1 Because of these minerals, the
leaves were effective in curing acne, scars, and other common skin problems.2 Annonaceous
acetogenins are waxy substances consisting of C32 or C34 long chain fatty acids which have
been combined with a 2-propanol unit at C-2 to form a lactone are only found in several genera
of the plant family, Annonaceae like guyabano trees. Its diverse bioactivities are as antitumor,
immunosuppressive, pesticidal, antiprotozoal, anti-feedant, anthelmitic, antibacterial and
antimicrobial agents.3
Staphylococcus epidermidis is a gram-positive and coagulase-negative staphylococci and
commonly present in human skin.4 .It is approximately 0.5 to 1.5 micrometers in diameter and
one of the most common pathogen that causes nosocomial infections (infections that are acquired
through the hospital) . Those most susceptible to these infections are drug users , newborns and
elderly with catheters. S. epididermis also causes infections to wounds. Infections and
4 | P a g e
-
7/28/2019 wpaper (1)
5/22
inflammation of wounds caused by the bacteria needs astringents with higher content and
solutions.
Most astringents are expensive and yet not effective. The pathogen like S. epidermidis
are still unaffected by these products. Due to this, the researcher will conduct this research to
determine the antibacterial activity of guyabano (Annona muricata) leaves extract to reduce skin
inflammation or infection and to contribute to the society a raw material which can be made as
an astringent product.
Statement of the Problem
This study was conducted to determine the effectiveness of guyabano (Annona muricata)
Leaves extract against the pathogenic bacteria S. epidermidis that is commonly found in the skin.
Specifically the proponent will answer the following problems and questions:
General Problem:
Is the guyabano leaves extract effective against the pathogenic bacteria
Staphylococcus epidermidis?
Specific Problem:
1. What concentration of guyabano extracts to distilled water will be the most effective in
killing the bacteria staphylococcus epidermidis?
a. 20 ml distilled water
b. 30 g guyabano leaves
c. 10 grams of guyabano leaves and 20 ml of water
d. 20 grams of guyabano leaves and 30 ml of water
5 | P a g e
-
7/28/2019 wpaper (1)
6/22
e. 30 grams of guyabanoleaves and 30 ml of water
f. 40 grams of guyabanoleaves and 30 ml of water
g. 50 grams of guyabanoleaves and 30 ml of water
Hypothesis
Is the guyabno leaves extract effective against the pathogenic bacteria Staphylococcus
epidermidis?
Operational Hypothesis
The guyabano leaves extract is effective against pathogenic bacteria Staphylococcus
epidermidis
Null Hypothesis
The Annona Muricata leaves extract is not effective in killing the pathogenic bacteria
Staphylococcus epidermidis.
Significance of the study
This study will be beneficial to the following:
Botany. This study provides prominent information about guyabano leaves. It may also lead to
the discovery of other species of the plants that can be used for further studies related to the
present studies
Department of Health.
They can endorse the use of this natural and herbal product in minimizing the cases of infection
caused by S. epidermidis in our country.
6 | P a g e
-
7/28/2019 wpaper (1)
7/22
Medicine and Health and other related fields.
The antibacterial substance of plant establishes other properties which may lead to further
discoveries of other biological and medical products and techniques which may be developed to
cure other diseases.
Other Researchers.
This may serve as a related study as a reference and guide in conducting further studies.
Scope and Limitations
This study will be conducted to determine if the Guyabano leaves extract is effective in
killing the bacteria Staphylococcus epidermidis. It is only concerned on one specific bacteria
Staphylococcus epidermidis and not any another bacteria found in the skin. This study will also
be conducted to determine the proportion of guyabano extract to distilled water that will be the
most effective in killing the bacteria.
Definition of Terms
1. Annona Muricata
-Annona muricata is a member of the family of Custard apple trees called Annonaceae
and a species of the genusAnnonaknown mostly for its edible fruits Anona.
-This is the raw material that will be used in the study
2. Staphylococcus epidermidis
-a common skin and mucosal inhabitant in humans and occasionally in animals living in
association with humans.
-This is the bacteria that will be acted by guyabano
7 | P a g e
http://en.wikipedia.org/wiki/Annonaceaehttp://en.wikipedia.org/wiki/Annonahttp://en.wikipedia.org/wiki/Annonahttp://en.wikipedia.org/wiki/Anonahttp://en.wikipedia.org/wiki/Annonaceaehttp://en.wikipedia.org/wiki/Annonahttp://en.wikipedia.org/wiki/Anona -
7/28/2019 wpaper (1)
8/22
3. Sensitivity Test
-a procedure which determines how responsive the subject is to a certain outside stimuli
- this procedure will be used to determine how effective is the proportion of leaves
extract to water in killing Staphylococcus epidermidis
4. Zone of inhibition
- The clear region around the paper disc saturated with anti microbial agent on the agar
surface.
-This is the one that is being measured to determine the effectiveness of the raw material.
8 | P a g e
-
7/28/2019 wpaper (1)
9/22
CONCEPTUAL FRAMEWORK
9 | P a g e
-
7/28/2019 wpaper (1)
10/22
END NOTES
1. Miss Pinky Soursop Medicinal Uses.2012. http://www.all-about-philippine-fruits-and-
herbs.com/index.html.
10 | P a g e
SET-UPS
SETUP 1
SETUP 2
SETUP 3
SETUP 4
SETUP 5
SETUP 6
SETUP 7
Replicate 1
Replicate 2Replicate 3
Replicate 1Replicate 2
Replicate 3
Replicate 1Replicate 2
Replicate 3
Replicate 1Replicate 2
Replicate 3
Replicate 1Replicate 2
Replicate 3
Replicate 1Replicate 2
Replicate 3
Replicate 1Replicate 2
Replicate 3
http://www.all-about-philippine-fruits-and-herbs.com/index.htmlhttp://www.all-about-philippine-fruits-and-herbs.com/index.htmlhttp://www.all-about-philippine-fruits-and-herbs.com/index.htmlhttp://www.all-about-philippine-fruits-and-herbs.com/index.html -
7/28/2019 wpaper (1)
11/22
2. Guyabano Herbal Medicine .Medicinal Health Guide.
2011.http://www.medicalhealthguide.com/herb/guyabano.htm
3. Muller,Viana. Graviola (Annona Muricata). 8 Jan 2010 . Whole World Botanicals.
http://wholeworldbotanicals.com/herbal_graviola
4. Staphylococcus epidermidis. 2010. Medpedia.inc.
http://wiki.medpedia.com/Staphylococcus_epidermidis.
Review of Related Literature
Annonaceous acetogenins
11 | P a g e
http://wholeworldbotanicals.com/herbal_graviolahttp://wholeworldbotanicals.com/herbal_graviola -
7/28/2019 wpaper (1)
12/22
Annonaceous acetogenins are powerful phytochemicals that are found in guyabano
plants.Many bioactive compounds and phytochemicals have been found in guyabano as scientists
have been studying its properties since the 1940's. Its many uses in natural medicine has been
validated by this scientific research. The earliest studies were between 1941 and 1962. Several
studies over the years have demonstrated that leaf, bark, root, stem and seed extracts of Graviola
are antibacterial in vitro against numerous pathogens and that the bark has antifungal properties.
Graviola seeds demonstrated active antiparasitic properties in a 1991 study, and a leaf extract
showed to be active against malaria in two other studies in 1990 and 1993.1. Annonaceous
acetogenins are only found in the Annonaceae family (to which Guyabano belongs). In general,
various Annonaceous acetogenins in the plant family have been documented with antitumorous,
antiparasitic, pesticidal, antiprotozoal, antifeedant, anthelmintic, and antimicrobial activities.
Guyabano
Guyabano is a small, upright evergreen tree, 5-6 m high, with large, glossy, dark green
leaves. It produces a large, heart-shaped, edible fruit that is 15-23 cm in diameter, is yellow-
green in color, and has white flesh inside. Guyabano is indigenous to most of the warmest
tropical areas in South and North America, including the Amazon and Asia. The fruit is sold in
local markets in the tropics, where it is called guanabana in Spanish-speaking countries and
graviola in Brazil. The fruit pulp is excellent for making drinks and sherbets and, though slightly
sour-acid, can be eaten.
All parts of the Guyabano tree are used in natural medicine in the tropics, including the
bark, leaves, roots, fruit, and fruit seeds. Different properties and uses are attributed to the
different parts of the tree. Generally, the fruit and fruit juice are taken for worms and parasites, to
cool fevers, as a lactagogue (to increase mother's milk after childbirth), and as an astringent for
12 | P a g e
-
7/28/2019 wpaper (1)
13/22
diarrhea and dysentery. The crushed seeds are used as a vermifuge and anthelmintic against
internal and external parasites, head lice, and worms. The bark, leaves, and roots are considered
sedative, antispasmodic, hypotensive, and nervine, and a tea is made
for various disorders toward those effects. Researchers verified Guyabano leaf's hypotensive
properties in rats again in 1991. Many bioactive compounds and phytochemicals have been
found in Guyabano, as scientists have been studying its properties since the 1940s. Its many uses
in natural medicine have been validated by scientific research. Several studies over the years
have demonstrated that leaf, bark, root, stem, and seed extracts of Guyabano are antibacterial in
vitro against numerous pathogens, and that the bark has antifungal properties. Guyabano seeds
demonstrated active antiparasitic properties in a 1991 study, and a leaf extract showed to be
active against malaria in two other studies (in 1990 and 1993). The leaves, root, and seeds of
Guyabano demonstrated insecticidal properties, with the seeds demonstrating strong insecticidal
activity in an early 1940 study.2.
Antibacterial
An antibacterial is an agent that inhibits bacterial growth or kills bacteria. The term is
often used synonymously with the term antibiotic(s); today, however, with increased knowledge
of the causative agents of various infectious diseases, antibiotic(s) has come to denote a broader
range ofantimicrobial compounds, including anti-fungal and other compounds.
The term antibiotic was first used in 1942 by Selman Waksman and his collaborators in journal
articles to describe any substance produced by a microorganism that is antagonistic to the growth
of other microorganisms in high dilution. This definition excluded substances that kill bacteria,
but are not produced by microorganisms (such as gastric juices and hydrogen peroxide). It also
excluded synthetic antibacterial compounds such as the sulfonamides. Many antibacterial
13 | P a g e
http://en.wikipedia.org/wiki/Bacteriahttp://en.wikipedia.org/wiki/Synonymhttp://en.wikipedia.org/wiki/Antimicrobialhttp://en.wikipedia.org/wiki/Anti-fungal_medicationhttp://en.wikipedia.org/wiki/Selman_Waksmanhttp://en.wiktionary.org/wiki/antagonismhttp://en.wikipedia.org/wiki/Gastric_juiceshttp://en.wikipedia.org/wiki/Hydrogen_peroxidehttp://en.wikipedia.org/wiki/Chemical_synthesishttp://en.wikipedia.org/wiki/Sulfonamide_(medicine)http://en.wikipedia.org/wiki/Bacteriahttp://en.wikipedia.org/wiki/Synonymhttp://en.wikipedia.org/wiki/Antimicrobialhttp://en.wikipedia.org/wiki/Anti-fungal_medicationhttp://en.wikipedia.org/wiki/Selman_Waksmanhttp://en.wiktionary.org/wiki/antagonismhttp://en.wikipedia.org/wiki/Gastric_juiceshttp://en.wikipedia.org/wiki/Hydrogen_peroxidehttp://en.wikipedia.org/wiki/Chemical_synthesishttp://en.wikipedia.org/wiki/Sulfonamide_(medicine) -
7/28/2019 wpaper (1)
14/22
compounds are relatively small molecules with a molecular weight of less than 2000 atomic
mass units.
With advances in medicinal chemistry, most of today's antibacterials chemically are
semisynthetic modifications of various natural compounds. These include, for example, thebeta-
lactam antibacterials, which include thepenicillins (produced by fungi in the genusPenicillium),
the cephalosporins, and the carbapenems. Compounds that are still isolated from living
organisms are the aminoglycosides, whereas other antibacterialsfor example, the
sulfonamides, the quinolones, and the oxazolidinonesare produced solely by chemical
synthesis. In accordance with this, many antibacterial compounds are classified on the basis of
chemical/biosynthetic origin into natural, semisynthetic, and synthetic. Another classification
system is based on biological activity; in this classification, antibacterials are divided into two
broad groups according to their biological effect on microorganisms: bactericidal agents kill
bacteria, andbacteriostatic agents slow down or stall bacterial growth3.
Staphylococcus Epidermidis
Staphylococcus epidermidis is a gram positive bacterium and one of 33 known species
belonging to the genus Staphylococcus. It is part of the human skin flora and consequently part
of the human microbiome. Although S. epidermidis is not usually pathogenic, patients with
compromised immune systems are often at risk for developing an infection. These infections can
be both nosocomial or community acquired, but they pose a greater threat to hospital patients. S.
epidermidis is also a major concern for people with catheters or other surgical implants because
it is known to causebiofilms that grow on these devices.4. When S. epidermidis causes illness in
humans, it most commonly results in many clinical conditions like blood stream infections,
Endocarditis, Cerebrospinal fluid (CSF) shunt infection, Peritoneal dialysis catheter infection,
14 | P a g e
http://en.wikipedia.org/wiki/Small_moleculehttp://en.wikipedia.org/wiki/Molecular_weighthttp://en.wikipedia.org/wiki/Atomic_mass_unithttp://en.wikipedia.org/wiki/Atomic_mass_unithttp://en.wikipedia.org/wiki/Medicinal_chemistryhttp://en.wikipedia.org/wiki/Semisynthetichttp://en.wikipedia.org/wiki/Beta-lactam_antibiotichttp://en.wikipedia.org/wiki/Beta-lactam_antibiotichttp://en.wikipedia.org/wiki/Penicillinhttp://en.wikipedia.org/wiki/Penicilliumhttp://en.wikipedia.org/wiki/Cephalosporinhttp://en.wikipedia.org/wiki/Carbapenemhttp://en.wikipedia.org/wiki/Aminoglycosidehttp://en.wikipedia.org/wiki/Sulfonamideshttp://en.wikipedia.org/wiki/Quinolonehttp://en.wikipedia.org/wiki/Oxazolidinonehttp://en.wikipedia.org/wiki/Biosynthesishttp://en.wikipedia.org/wiki/Bactericidehttp://en.wikipedia.org/wiki/Bacteriostatic_agenthttp://en.wikipedia.org/wiki/Gram_positivehttp://en.wikipedia.org/wiki/Staphylococcushttp://en.wikipedia.org/wiki/Immune_systemhttp://en.wikipedia.org/wiki/Nosocomialhttp://en.wikipedia.org/wiki/Cathetershttp://en.wikipedia.org/wiki/Biofilmshttp://wiki.medpedia.com/Endocarditishttp://en.wikipedia.org/wiki/Small_moleculehttp://en.wikipedia.org/wiki/Molecular_weighthttp://en.wikipedia.org/wiki/Atomic_mass_unithttp://en.wikipedia.org/wiki/Atomic_mass_unithttp://en.wikipedia.org/wiki/Medicinal_chemistryhttp://en.wikipedia.org/wiki/Semisynthetichttp://en.wikipedia.org/wiki/Beta-lactam_antibiotichttp://en.wikipedia.org/wiki/Beta-lactam_antibiotichttp://en.wikipedia.org/wiki/Penicillinhttp://en.wikipedia.org/wiki/Penicilliumhttp://en.wikipedia.org/wiki/Cephalosporinhttp://en.wikipedia.org/wiki/Carbapenemhttp://en.wikipedia.org/wiki/Aminoglycosidehttp://en.wikipedia.org/wiki/Sulfonamideshttp://en.wikipedia.org/wiki/Quinolonehttp://en.wikipedia.org/wiki/Oxazolidinonehttp://en.wikipedia.org/wiki/Biosynthesishttp://en.wikipedia.org/wiki/Bactericidehttp://en.wikipedia.org/wiki/Bacteriostatic_agenthttp://en.wikipedia.org/wiki/Gram_positivehttp://en.wikipedia.org/wiki/Staphylococcushttp://en.wikipedia.org/wiki/Immune_systemhttp://en.wikipedia.org/wiki/Nosocomialhttp://en.wikipedia.org/wiki/Cathetershttp://en.wikipedia.org/wiki/Biofilmshttp://wiki.medpedia.com/Endocarditis -
7/28/2019 wpaper (1)
15/22
Urinary Tract Infection, Infection inprosthetic joints, Infection of vascular grafts (tubes inserted
into blood vessels to bypass areas of blockage or damage), Infection in newborn children and
Infection of breast implants5.. The natural environment ofS. epidermidis is the human body and
usually originates from disease. Since the bacteria usually lives on the skin and nares of all
human beings and is a nosocomial pathogen, it is imporant to be able to identify the specific
strains. S. epidermidis is the most common staphylococcus on the human skin. In addition, S.
epidermidis also covers 90%-100% staphylococci from the nares when S. aureus is not present.
When S. aureus is present the S. epidermidis amount decreases drastically. The formation of
biofilm allows S. epidermidis to attach and grow on biomedical devices and be released into the
blood to infect new areas. The increase use of intravascular catheters has caused a similar
increase in S. epidermidis infections. The increase causes a problem since S. epidermidis is
resistant to methicillin and all penicillins, penems, carbapanems, and cephalosporins which are
commonly used antibiotics. S. epidermidis has also been found to be more resistant to antibiotics
than other species
Although there are no one specific viruelence factors ofS. epidermidis, the ability to form
biofilm is one of the virulence factors. The biofilm allows the bacteria cells to adhere to an inert
or living areas .When a biofilm has formed it becomes harder to treat since the cells inside the
biofilm are guarded from antibiotics and the immune system . Biofilm also releases a host
immune response to antigens which prevents the removal of the biofilm and may also result in
tissue damage. The bacteria can be released into the blood from biofilms and start new infections
by attachment to the medical devices, thus devices will be required to be removed .
Some preventive strategies for infections are to provide prohylatic antibiotic therapy to cover
surgical insertions from temporary intravascular devices. There are also reports that warn to not
15 | P a g e
http://wiki.medpedia.com/Urinary_Tract_Infectionhttp://wiki.medpedia.com/Joint_Replacementhttp://wiki.medpedia.com/Urinary_Tract_Infectionhttp://wiki.medpedia.com/Joint_Replacement -
7/28/2019 wpaper (1)
16/22
use antibiotic prohylaxis, especially vancomycin for dialysis. Aseptic techniques should be used
for catheter infections to prevent contamination. New techniques concentrate on physical
electrical barriers for colonization and using biomaterials with antimicrobial agents already
inside. However these new methods have not been tried in the clinical settings6..
Biofilms
Biofilms are communities of microbial cells that grow on living or inert surfaces and
surround themselves with secreted polymers. Many bacterial species like staphylococcus
epidermidis form biofilms, and their study has revealed them to be complex and diverse. The
structural and physiological complexity of biofilms has led to the idea that they are coordinated
and cooperative groups, analogous to multicellular organisms.7.. Biofilms are usually found on
solid substrates submerged in or exposed to an aqueoussolution, although they can form as
floating mats on liquid surfaces and also on the surface of leaves, particularly in high humidity
climates. Given sufficient resources for growth, a biofilm will quickly grow to be macroscopic
(visible to the naked eye). Biofilms can contain many different types of microorganism,
e.g. bacteria,archaea,protozoa, fungi and algae; However, some organisms will form single-
species films under certain conditions.7. Biofilms have been found to be involved in a wide
variety of microbial infections in the body, by one estimate 80% of all infections. Infectious
processes in which biofilms have been implicated include common problems such asurinary
tract infections, catheterinfections, middle-ear infections, formation ofdental plaque,gingivitis,
coating contact lenses, and less common but more lethal processes such asendocarditis,
infections in cystic fibrosis, and infections of permanent indwelling devices such as
jointprostheses andheart valves. More recently it has been noted that bacterial biofilms may
16 | P a g e
http://en.wiktionary.org/wiki/substratehttp://en.wikipedia.org/wiki/Aqueoushttp://en.wikipedia.org/wiki/Solutionhttp://en.wikipedia.org/wiki/Bacteriahttp://en.wikipedia.org/wiki/Archaeahttp://en.wikipedia.org/wiki/Protozoahttp://en.wikipedia.org/wiki/Fungushttp://en.wikipedia.org/wiki/Algaehttp://en.wikipedia.org/wiki/Urinary_tract_infectionshttp://en.wikipedia.org/wiki/Urinary_tract_infectionshttp://en.wikipedia.org/wiki/Urinary_tract_infectionshttp://en.wikipedia.org/wiki/Urinary_catheterizationhttp://en.wikipedia.org/wiki/Otitis_mediahttp://en.wikipedia.org/wiki/Dental_plaquehttp://en.wikipedia.org/wiki/Gingivitishttp://en.wikipedia.org/wiki/Gingivitishttp://en.wikipedia.org/wiki/Contact_lenseshttp://en.wikipedia.org/wiki/Endocarditishttp://en.wikipedia.org/wiki/Endocarditishttp://en.wikipedia.org/wiki/Cystic_fibrosishttp://en.wikipedia.org/wiki/Prosthesishttp://en.wikipedia.org/wiki/Heart_valve_prosthesishttp://en.wikipedia.org/wiki/Heart_valve_prosthesishttp://en.wiktionary.org/wiki/substratehttp://en.wikipedia.org/wiki/Aqueoushttp://en.wikipedia.org/wiki/Solutionhttp://en.wikipedia.org/wiki/Bacteriahttp://en.wikipedia.org/wiki/Archaeahttp://en.wikipedia.org/wiki/Protozoahttp://en.wikipedia.org/wiki/Fungushttp://en.wikipedia.org/wiki/Algaehttp://en.wikipedia.org/wiki/Urinary_tract_infectionshttp://en.wikipedia.org/wiki/Urinary_tract_infectionshttp://en.wikipedia.org/wiki/Urinary_catheterizationhttp://en.wikipedia.org/wiki/Otitis_mediahttp://en.wikipedia.org/wiki/Dental_plaquehttp://en.wikipedia.org/wiki/Gingivitishttp://en.wikipedia.org/wiki/Contact_lenseshttp://en.wikipedia.org/wiki/Endocarditishttp://en.wikipedia.org/wiki/Cystic_fibrosishttp://en.wikipedia.org/wiki/Prosthesishttp://en.wikipedia.org/wiki/Heart_valve_prosthesis -
7/28/2019 wpaper (1)
17/22
impair cutaneous wound healing and reduce topical antibacterial efficiency in healing or treating
infected skin wounds.
It has recently been shown that biofilms are present on the removed tissue of 80% of
patients undergoing surgery for chronicsinusitis. The patients with biofilms were shown to have
been denuded ofcilia and goblet cells, unlike the controls without biofilms who had normal cilia
and goblet cell morphology. Biofilms were also found on samples from two of 10 healthy
controls mentioned. The species of bacteria from interoperative cultures did not correspond to
the bacteria species in the biofilm on the respective patient's tissue. In other words, the cultures
were negative though the bacteria were present.
Biofilms can also be formed on the inert surfaces of implanted devices such as catheters,
prosthetic cardiac valves and intrauterine devices.
New staining techniques are being developed to differentiate bacterial cells growing in living
animals, e.g. from tissues with allergy-inflammations . 8
END NOTES
17 | P a g e
http://en.wikipedia.org/wiki/Sinusitishttp://en.wikipedia.org/wiki/Sinusitishttp://en.wikipedia.org/wiki/Ciliumhttp://en.wikipedia.org/wiki/Goblet_cellshttp://en.wikipedia.org/wiki/Goblet_cellshttp://en.wikipedia.org/wiki/Sinusitishttp://en.wikipedia.org/wiki/Ciliumhttp://en.wikipedia.org/wiki/Goblet_cells -
7/28/2019 wpaper (1)
18/22
1. Annonaceous Acetogenins. 2011.Pub Med, National Library of Medicine.
http://www.immupro.com/graviola.htm.
2. Guyabano(Annona Muricata). http://guyabano.com/Page_2.html.
3. Antibacterial.19 Nov 2012.Wikimedia Foundation, Inc.
http://en.wikipedia.org/wiki/Antibacterial
4. Staphylococcus Epidermidis 21 February 2013. Wikimedia Foundation, Inc.
http://en.wikipedia.org/wiki/Staphylococcus_epidermidis
5. Staphylococcus epidermidis. 2010. Medpedia.inc.
http://wiki.medpedia.com/Staphylococcus_epidermidis.
6. Staphylococcus Epidermidis. 22 April 2011 . Microbewiki Foundation Inc.
http://microbewiki.kenyon.edu/index.php/Staphylococcus_epidermidis.
7. Biofilm Bacteria 2 Jan 2012. Autoimmunity Research Foundation.
http://mpkb.org/home/pathogenesis/microbiota/biofilm
8. Biofilm. 23 Feb 2013 . Wikimedia Foundation Inc.
http://en.wikipedia.org/wiki/Biofilms
Methodology
18 | P a g e
http://en.wikipedia.org/wiki/Antibacterialhttp://wiki.medpedia.com/Staphylococcus_epidermidishttp://en.wikipedia.org/wiki/Antibacterialhttp://wiki.medpedia.com/Staphylococcus_epidermidis -
7/28/2019 wpaper (1)
19/22
Methods of research
The methods of research used in the study are experimental and observation method.
Experimental method will be used to test the effectiveness of guyabano (Anonna muricata)
leaves against the bacteria Staphylococcus epidermidis and observation method to determine the
range of the zone of inhibition of the proportions. This will determine what proportion is the
most effective in killing the pathogenic bacteria Staphylococcus epidermidis.
The study is mainly PHBA(Potentially Hazardous Biological Agents) because it focuses
in killing pathogenic bacteria that can harm living organism. Thus this study focuses in killing
S.epidermidis using the extract from guyabano leaves
Methods of Collecting Data
The methods used for collecting data is experimental method and library method . The
experimental method used in the study is concerned with the extraction of the guyabano leaves
against S. epidermidis. It is more appropriate to use the experimental method in collecting
specific and reliable data. Library method is gathering information through books, literatures and
articles and is used to find related literatures to help answers of the specific problems.
Statistical Treatment
The statistical treatment will be used to show the results of experimentations through
mathematical graphs.Each proportion has 3 replicates in which we will get the mean and
compare which proportion more effective.
Materials
19 | P a g e
-
7/28/2019 wpaper (1)
20/22
To get the extract, the researcher will be utilizing a total of 180 grams guyabano leaves,
and 120 mL distilled water. Mortar and pestle, beaker, cheese cloth, weighing scale and plastics
bags will also be used.
Procedure: Extraction
Concentration 1 (negative control)
First, the 20ml distilled water will be measured using the beaker. Then it will be poured
into the small bottle using a funnel and will be labeled as CONCENTRATION 1.
Concentration 2 (30 grams of guyabano leaves) (Pure extract)
First, the 30g of guyabano leaves will be labeled and will be placed inside a plastic bag
and pounded using the mortar and pestle. Using the cheese cloth, the pounded leaves will be
squeezed until the pure extract is obtained. The extract will be then placed in test tube B and will
be transferred into a small bottle and will be labeled as CONCENTRATION 2.
Concentration 3 (10 grams of guyabano leaves and 20 ml of water)
The 10g of guyabano leaves will be labeled and placed inside a plastic bag. Then, a 50ml
beaker will be filled up with 20 ml of distilled water and will be poured in the plastic bag with
the leaves in it and will be pounded. To get the extract, same process in concentration 2 will be
used. The extract will be then placed in test tube C and will be transferred into a small bottle and
will be labeled as CONCENTRATION 3.
Concentration 4 (20 grams of guyabano leaves and 30 ml of water)
First, the 20g of guyabano leaves will be labeled and placed inside a plastic bag. Then, a
50ml beaker will be filled up with 30 ml of distilled water and will be poured in the plastic bag
20 | P a g e
-
7/28/2019 wpaper (1)
21/22
with the leaves in it and will be pounded. To get the extract, same process in concentration 2 will
be used. The extract will be then placed in test tube D and will be transferred into a small bottle
and will be labeled as CONCENTRATION 4.
Concentration 5 (30 grams of guyabanoleaves and 30 ml of water)
First, the 30g of guyabanoleaves will be labeled and placed inside a plastic bag. Then, a
50ml beaker will be filled up with 20 ml of distilled water and will be poured in the plastic bag
with the leaves in it and will be pounded. To get the extract, same process in concentration 2 will
be used. The extract will be then placed in test tube E and will be transferred into a small bottle
and will be labeled as CONCENTRATION 5.
Concentration 6 (40 grams of guyabanoleaves and 30 ml of water)
First, the 40g of guyabano leaves will be labeled and placed inside a plastic bag. Then, a
50ml beaker will be filled up with 20 ml of distilled water and will be poured in the plastic bag
with the leaves in it and will be pounded. To get the extract, same process in concentration 2 will
be used. The extract will be then placed in test tube F and will be transferred into a small bottle
and will be labeled as CONCENTRATION 6.
Concentration 7 (50 grams of guyabanoleaves and 30 ml of water)
First, the 50g of guyabanoleaves must bewill be labeled and placed inside a plastic bag.
Then, a 50ml beaker will be filled up with 20 ml of distilled water and will be poured in the
plastic bag with the leaves in it and will be pounded. To get the extract, same process in
concentration 2 will be used. The extract will be then placed in test tube G and will be
transferred into a small bottle and will be labeled as CONCENTRATION 7.
21 | P a g e
-
7/28/2019 wpaper (1)
22/22
22 | P a g e