Documenti di Didattica
Documenti di Professioni
Documenti di Cultura
CHAPTER 1
conducting the study. It includes the introduction, statement of the problem, hypothesis,
significance of the study, scope and delimitation, and the definition of terms.
Introduction
In traditional herbal medicine, plants have been used for many years.
Therefore, plants have attained status of natural source of new and potent antimicrobial
agents. Medicinal plants are used as ethnomedicine in different countries around the
world and are source of natural products providing unlimited opportunity for new drugs
Traditional healers have long used plants to prevent or cure infectious conditions.
Plants are rich in a wide variety of secondary metabolites, such as chave antimicrobial
properties(Cowan, 2011).
metabolomics are nowadays applied in medicinal plant research and contribute to the
Anacardiaeceae which consists of about sixty genera and six hundred species.
Conventionally, the mango plant has several biological properties as like to treat mouth
sore throat and scurvy. It is also a source of vitamin A when eaten in combinations with
salt and honey for the treatment of blood disorders. Ground seeds and leaves have been
used as remedy for diabetes, as colic and to treat irritations, scorpion and bee
in order to assist a physician in selecting treatment options for his or her patients. The
antimicrobial impregnated filter paper disks. The presence or absence of growth around
the disks is an indirect measure of the ability of that compound to inhibit that organism
(Hudzicki, 2010).
osteoarticular, skin and soft tissue, pleuropulmonary, and device-related infections. The
past 2 decades have witnessed two clear shifts in the epidemiology of S.
particularly seen in infective endocarditis and prosthetic device infections, and second, an
epidemic of community-associated skin and soft tissue infections driven by strains with
certain virulence factors and resistance to β-lactam antibiotics( Davis et al., 2015).
(Mangifera indica) leaf extract which is abundant in our country, cheaper, eco-friendly,
3
and less hazardous. This study will give highlights on determining the capability of
Mango (Mangifera indica) leaf extract as an antimicrobial agent and its importance in
Staphylococcus aureus.
antimicrobial agents?
3.) Which is the most effective menstruum used for the extraction of Mango
4.) What is the significance of the study among the researchers in the field of
Medical Technology?
Hypothesis
For Students.The study will enlighten the minds of the students aboutMango (Mangifera
indica) leaf extract as an antimicrobial agent. It will serve as their reference or guide in
creating their own study. It will also help the students who are taking Medical
For Medical Technologists.The Medical Technologist will benefit on the said study in
the following terms: health – it is safer to use because of its organic components; Cost-
cutting – it is cheaper than the commercial antimicrobial products present in the market.
For Manufacturers. The manufacturers will benefit in the study because it will help
is abundant in our country or anywhere and it is less toxic than the chemical used in
making antimicrobial products. It will also give them the idea that .Mango (Mangifera
indica), is not only just a fruit we often buy at the market but also it can be use in the
For the Community.The study will also be beneficial in the community because Mango
(Mangifera indica) can be purchased in any market that would give them the opportunity
For the Future Researchers. The outcome of the study will be beneficial for the future
researchers. It would be a great help for the future researchers to be more creative and
5
more productive in conducting a study. The research will give highlight on how
The scope of the study was solely focused on the antimicrobial capability of
Mango (Mangifera indica) leaf extract against S. aureus. The researcher used Disk
diameter of inhibitory zones of the Mango (Mangifera indica) leaf extract to determine its
Definition of Terms
The following terms are defined conceptually and operationally for better
Antimicrobial. An agent that stops the growth of S. aureus. In this study, the
Mango (Mangifera indica) leaf extract was tested for its antimicrobial action.
Capability. This refers to the ability Mango (Mangifera indica) leaf extract as an
antimicrobial agent.
concentrated form. In this study, maceration method was used to produce the optimum
In this study, Mango (Mangifera indica) leaf extract was tested if it is an effective
antimicrobial agent.
motile cocci and usually found in grape-like clusters. It is found in the environment and is
a normal human flora, located on the skin and mucous membranes of most healthy
wide variety of clinical manifestations. In this study, it is one of the bacteria that was
CHAPTER 2
This chapter includes information from the literatures that are related to this
research and studies done by other researchers that will correlate for the further
Related Literature
Ayurvedic and indigenous medical systems for over 4000 years. Mangoes belong to
properties are attributed to different parts of mango tree. Mango is one of the most
popular of all tropical fruits. Mangiferin, being a polyphenolic antioxidant and a glucosyl
In recent years, there has been a dramatic increase in number of reported poisoning
activity, smoking, fermentation, and adding antimicrobial agents, have been used to
8
prevent spoilage of foods. The addition of antimicrobial agents has been a particularly
antimicrobial agents, those from natural sources are acceptable to consumers. Phenolic
compounds, tannins and flavonoids in mango seed are found to be responsible for
microbial; growth and by avoiding oxidative phosphorylation. So mango seed kernel oil
is used to kill abdominal worm and also given as a cure for vomiting, diarrhea and
hyperacidity. (Kittiphoom,2011)
Infectious diseases are one of the main causes of morbidity and mortality worldwide.
Nature is a generous source of compounds, with the potential to treat diseases, including
infectious diseases. Studies exploiting the mechanism of action and the structure-activity
aspects of these natural compounds may provide both additional antimicrobial leads and
drugs, and also significant insight into potential possibilities to overcome the
Plants are used for the treatment of diseases and novel drugs continue to be
developed through research from plants. There are more than 20,000 plant species
used in traditional medicines and they are prospective reservoirs for new
because their phytochemicals may lead to new drug discoveries.For several years,
there has been an increased interest in the study of different extracts obtained from
search for new antimicrobial agents is an important line of research because of the
and have shown an evolution in resistance to various pharmaceutical drugs. Because the
levels of resistant pathogenic microorganisms are on the rise, novel and effective
antimicrobial agents are needed to contain this epidemic. Natural products obtained from
various plant species have been used for maintaining health as evidenced by their
Plants have been used as food, spice, and medicine since ancient time. Much
before the discovery of the existence of microorganisms, plants were an integrated part of
the traditional medicine of many communities used for the treatment of various infectious
In the recent years, research on medicinal plants has attracted a lot of attentions
globally. Large body of evidence has accumulated to demonstrate the promising potential
treatment of human diseases. Plants are rich in a wide variety of secondary metabolites
such as tannins terpenoids, alkaloids, flavonoids, etc, which have been found in vitro to
skin and skin structure infections as well as osteoarticular infections in the pediatric
10
mobile genetic elements that transfer resistance and virulence from other bacterial species
and staphylococcal strains. External pressures (namely, overuse of antibiotics) can also
part of normal human flora. A common body area of S aureus colonization is the anterior
nares. Colonization is a risk factor for infection, and infection often occurs with the
Related Studies
A. Foreign
(Mangiferaindica) stem bark extracts was determined using disk diffusion, agar and broth
dilution methods. In disk diffusion method, inhibition zone sizes were used to determine
the susceptibility of S. aureus to the extracts. The results showed that the stem-bark
showed the highest inhibition zone diameter of 25 mm, followed by ethyl acetate, water
and hexane extracts with inhibition zone diameter of 22 mm, 14 mm and 10 mm,
concentration dependent, in agar and broth dilution methods. The plant extracts were
shown to have a MIC range of 0.62 mg/ml to 4.17 mg/ml, in agar dilution method.
Results from the broth dilution method had a MIC range of 0.16 mg/ml to 1.25 mg/ml.
11
The control (ampicillin) was however, more effective than plant extracts since only a
concentration of 0.03 mg/ml in agar dilution and 0.001 mg/ml in broth dilution method
were effective to inhibit the growth of S. aureus. The extracts were shown to be
presence of phyto-compounds such as alkaloids and tannins which are known to inhibit
bacterial growth by different mechanisms from those of synthetic drugs. These phyto-
Rawal et al (2010), Basella rubra were evaluated by measuring the inhibition zones using
Disk Diffusion method. The test organisms are Staphylococcus aureus, Escherichia coli,
Leaves were shade dried, powdered and then extracted with water, 95% ethanol and
petroleum ether for 48 hours using Soxhlet apparatus. The filterates were collected and
evaporated to dryness under reduced pressure using a Rota evaporator. The dried extracts
were stored in dry sterilized small containers at 4˚C. The ethanolic extract showed
aqueous extract (13.4±1.2 mm) and petroleum ether (5.6±0.62 mm) at a concentration of
Synthesis
(Mangifera indica) has an antimicrobial activity against bacterial cells. The study tested
Mango (Mangifera indica) plant for its antimicrobial property same with this study. The
study conducted by Takudzwa et al, used disk diffusion method on agar and broth, while
this study only used agar diffusion method. Takudzwa et al used methanol, water ethyl
acetate, and hexane as extracting solvents while this study used methanol, ethanol, and
water. The study used dried mango stem bark while this study used fresh mango leaves.
In the study conducted by Rawal et al (2010), they used diffusion method same with this
study. And also they used ethanolic, aqueous, and petroleum ether as an extracting
solvent. Soxhlet apparatus were used to obtain the extract while in this study maceration
Local
Bacteria”. The guava leaves were extracted in four different solvents of increasing
polarities (hexane, methanol, ethanol, and water). The efficacy of these extracts was
tested against those bacteria through a well-diffusion method employing 50 μL leaf-
extract solution per well. According to the findings of the antibacterial assay, the
methanol and ethanol extracts of the guava leaves showed inhibitory activity against
gram-positive bacteria, whereas the gram-negative bacteria were resistant to all the
solvent extracts. The methanol extract had an antibacterial activity with mean zones of
13
inhibition of 8.27 and 12.3 mm, and the ethanol extract had a mean zone of inhibition of
utilized using the transect-plot method. Ethnomedicinal uses and the mode of
ethnomedicinal plants: Crinum asiaticum L., Pavettaindica L., Bauhinia purpurea L.,
Mollugo pentaphylla L., and Cinnamomummercadoi S. Vidal were selected for the anti-
microbial test against cultured bacterial strains; the Escherichia coli and Staphylococcus
aureus using the disc diffusion method. The mode of preparation of the extracts follows
the traditional method of the indigenous people. Chloramphenicol, on the other hand, was
used as positive control. Thirty-three out of 61 identified species were locally claimed to
have medicinal value and are known to cure stomach disorders, poisoning, bleeding,
cough, fever and wounds, among others. Antimicrobial test showed greater inhibitory
Extracts of Twelve Common Medicinal Plants from the Philippines”. The antibacterial
twelve common medicinal plants from the Philippines obtained through pounding and
solvent extraction was evaluated using disc Agar diffusion. The microorganisms tested
aeruginosa. Susceptibility test results showed that different extracts (1000 mcg/disc) from
ten plants inhibited growth with a mean zone of inhibition range of 15 to 30 mm against
S. aureus, E. coli, and P. aeruginosa. The common medicinal plants which showed highly
(Moringa), Carmona retusa (Wild tea), Citrus aurantifolia (Lemon), Citrus sinensis
(Orange), Allium sativum (garlic), and Allium cepa (onion). Preliminary phytochemical
and steroids/terpenes. The results suggest that the different plant extracts contain
with very strong antibacterial activity and validates the ethno-medical use in the
treatment of bacterial skin diseases and other forms of bacterial infections. However,
many reputed plant antibacterials have very variable activities depending on certain
conditions.
Synthesis
Gram-Positive Bacteria”, guava leaf extracts was tested for its effectiveness as an
antimicrobial agent. The extraction method was also carried out through maceration
technique (less the drying up of leave). The similarity of their study from the researcher’s
study is, the researcher also used maceration technique to obtain the extract of the subject
which is Mango ( Mangifera indica) leaves. The study used hexane, methanol ,ethanol
15
and, water were used as an extracting solvent while this study used ethanol, methanol,
and water.
extracted using traditional method, by boiling the leaves in tap water, while in this study
the researcher used maceration technique to extract Mango (Mangifera indica) leaves.
The similarity of their study to the researcher’s study is, they used the different subjects
but the same purposes, Bandibas et al., used the leaves of five ethnomedicinal plants:
Crinum asiaticum L., Pavettaindica L., Bauhinia purpurea L., Mollugo pentaphylla L.,
and Cinnamomummercadoi S., while the researchers used Mango (Mangiferaindica) leaf
extract as an antimicrobial and the study afore mentioned used positive control same with
this study.
Common Medicinal Plants from the Philippines”. conducted by Penecilla et al., (2011),
the twelve medicinal plant extract were obtained using maceration technique and it used
disk diffusion method to measure the susceptibility of plant extracts against the bacterial
cells being tested same methods used in this study And also both studies utilized
controls.
Conceptual Framework
information that the researcher gathered, foreign and local studies and literature, proves
bacterial cells.
In this study, the researcher used disk diffusion method, which is the most common
technique in extracting Mango (Mangifera indica) leaves, 50% ethanol, 50% methanol,
and water were used as an extracting solvent. The researchers aim to determine the if
aureus.
Paradigm
Mango (Mangiferaindica)
leaf extracts Resistant/Susceptible
Figure 1 shows the scope of the the study to determine if Mango (Mangifera
indica) leaf extract can inhibit Staphylococcus aureus. The researchers used Mango
17
(Mangifera indica) leaf extract as the experimental variable, while the erythromycin had
been used as a control variable. Maceration and kirby bauer disk diffusion are utilized.
produced.
CHAPTER 3
RESEARCH METHODOLOGY
This chapter presents the method and procedure that will employed in this research.
It incorporates the research design, techniques, instruments and data gathering procedure.
Research Design
(Mangiferaindica)leaf extract was used as an experimental variable and subjected for its
Research Subjects
The researcher used Mango (Mangiferaindica) leaf extract as the main subject and
it was used to test its sensitivity against S. aureus. Mueller Hinton Agar was used to test
the subject’s antimicrobial capability against S. aureus. The researchers used 3 MHA
Research Locale
extract was subjected for identification tests to prove the presence of the active
18
constituents. The ampoule which contains the S. aureus was gathered from the Medical
technology department laboratory, are propagate for culture and sensitivity testing. The
tests was be conducted at the laboratory of Central Luzon Doctors’ Hospital Educational
Research Instruments
Materials Function
19
Alcohol lamp for the sterilization of inoculating loop
Gathering
Analytical balance for weighing of Mango (Mangiferaindica)
Procedure
leaves and reagent for preparing agar plate.
Phase
Beaker Used to contain the pounded mango leaves
1: Collection
soaked in different menstruum.
and
Mueller hinton agar plate serves as a medium for antimicrobial
Preparation
susceptibility testing.
of Plant
collected
Filter paper impregnated with Mango (Mangifera
from the
indica) leaf extract and was placed on a
Mango
Mueller-Hinton (MH) agar plate,
(Mangifera
Funnel used when transferring liquid into a narrow
indica) trees.
mouthed container
Whole leaves
growth.
the medium.
20
then crush using mortar and pestle. Using beakers, crushed leaves was weighed 10 grams,
then soaked to 50% ethanol, 50% menthanol, and water respectively (1:1). Then covered
with parafilm to avoid evaporation and contamination for 24 hours at room temperature.
After 24 hours of soaking in different menstruum, the mixtures was filtered using filter
paper. The residues was discarded while the filtrates was collected.
Technolody Department laboratory. Collected isolates was sub-cultured for 24 hours and
The researchers went to Central Luzon State University to authenticate the plant
Disks was prepared by the use of puncher to punch the filter paper into pieces.
Each measures 6 mm. The disks were soaked with Mango (Mangiferaindica) leaf extract
for 30 minutes.
Sensitivity Method
Inoculation of Plates
21
2. Rotate the swab against the side of the tube (above the fluid level) using firm pressure,
3. Inoculate the dried surface of a MH agar plate by streaking the swab three times over
the entire agar surface; rotate the plate approximately 60 degrees each time to ensure an
4. Rim the plate with the swab to pick up any excess liquid.
6. Leaving the lid slightly ajar, allow the plate to sit at room temperature at least 3 to 5
minutes, but no more than 15 minutes, for the surface of the agar plate to dry before
1. Place the appropriate antimicrobial-impregnated disks on the surface of the agar, using
either forceps to dispense each antimicrobial disk one at a time, or a multidisk dispenser
b. Place the dispenser over the agar plate and firmly press the plunger once to
c. Lift the dispenser off the plate and using forceps sterilized by either cleaning
them with an alcohol pad or flaming them with isopropyl alcohol, touch each disk on the
plate to ensure complete contact with the agar surface. This should be done before
replacing the petri dish lid as static electricity may cause the disks to relocate themselves
d. Do not move a disk once it has contacted the agar surface even if the disk is not
in the proper location, because some of the drug begins to diffuse immediately upon
e. To add disks one at a time to the agar plate using forceps, place the MH plate
on the template. Sterilize the forceps by cleaning them with a sterile alcohol pad and
allowing them to air dry or immersing the forceps in alcohol then igniting.
f. Using the forceps carefully remove one disk from the cartridge.
g. Partially remove the lid of the petri dish. Place the disk on the plate over one of
the dark spots on the template and gently press the disk with the forceps to ensure
complete contact with the agar surface. Replace the lid to minimize exposure of the agar
h. Continue to place one disk at a time onto the agar surface until all disks have
been placed.
2. Once all disks are in place, replace the lid, invert the plates, and place them in a 35°C
Interpretation of Results
23
extracts).
dosage of a drug.
Statistical Treatment
(Mangiferaindica) leaf extract against S. aureus, the researchers encoded the result in a
tabular form to compare the inhibitory activity of Mango (Mangiferaindica) leaf extract
against S. aureus.