Professional Documents
Culture Documents
CHAPTER I
THE PROBLEM AND SCOPE
Introduction
With the uprising population and commodities in the community, morbidity and
mortality: it is hard to avoid since the people in the society are also experiencing
phenomenal calamities like flood. Floods may carry microorganisms which can
contribute to poor wound healing.
The researchers conducted this study since the Carabao Grass (Paspalum
conjugatum) is known to have an antimicrobial property and is widely spread in local
areas and in most communities in Philippines.
In addition, the researchers want to help minimize the expenses in health services
most especially to the medications being bought. There are number of people living in far
flung areas that are unfamiliar to this kind of grass which can be a possible alternative
antimicrobial regimen. This research aims to prove that Carabao Grass (P.conjugatum)
has an antimicrobial agent. Through this, the researchers will be of help in disseminating
information to the people not only in remote areas but also to those people living in the
community.
The researchers aim to know more about the phytochemical analysis that are
found in Carabao Grass (P.conjugatum) and its potential action as an antimicrobial agent.
People are not aware about the idea of Carabao Grass as an antimicrobial agent (Ebadi,
2007) and through this research; people in the community are beneficial with regard to
the availability of resources.
Theoretical Framework
The study consists of antimicrobial assay, which then would determine the
attribute of P. conjugatum.
The researcher must endeavor and analyze things to equipped themselves and be
able to discover the significance of the study.
The Kirby-Bauer agar diffusion method of antimicrobial assay is a convenient and
efficient procedure to conduct the study. It is placing of filter paper disk with a
concentration of treatment and control extract. The diffusion will depend on the agar
properties and the molecular weight of the filter paper disk.
Antimicrobial activity may be observed based on the presence of the zones of
inhibition within the span of 24 hours. The slower the diffusion, the larger the molecular
weight present in the filter disk. The size of the inhibition zone will depend on the depth
of the agar.
There are some theories that can support the study of the researchers. Theories
like the Environmental Model. It focuses on the manipulation of the availability of
resources in the environment.
It is in this theory that focusing on changing and manipulating the environment in
order to put the patient in best possible conditions for nature to act. The said study has
relation to this theory for it benefits the community to lessen their expenses in health
services and can be an alternative antimicrobial regimen for people living far away from
health centers and pharmacies.
P. conjugatum as a promising
Institute of Technology (MSU-IIT) checked it. The preparation of the extract was in the
College of Nursing Microbiology Laboratory in Mindanao State University-Iligan
Institute of Technology (MSU-IIT), Iligan City. The stem leaves and roots were extracted
prior to assay.
Definition of Terms
Antimicrobial is an agent that kills microorganism or inhibits their growth.
Antimicrobial Assay- a method for quantitatively determining the concentration of an
antibiotic by its effect in inhibiting the growth of a susceptible microorganism.
Bacteria are microscopic living organism, usually one-celled, that can be found
everywhere.
Kirby-Bauer agar diffusion method -is a test which uses antibiotic-impregnated wafers
to test whether bacteria are affected by antibiotics
Minimum inhibition concentration is the lowest concentration of an antimicrobial
that will inhibit the visible growth of a microorganism after overnight incubation.
Mueller Hinton Agar is a microbial growth medium that is commonly used for
antibiotic susceptibility testing.
Zone of inhibition is an area of growth inhibition around a point source, within a
cultured organism on a solid medium, due to the action of growth-inhibitory substance
such as antimicrobial present at the source.
CHAPTER II
REVIEW OF RELATED LITERATURE
For over a thousand years, human beings primary sources for medicine are the
plants, until now people are developing newer medicines that are basically derived from
plants. Then and now has no apparently significant changes that are worthy to say that
people no longer use plants as a one of sources for medicines (Kala, 2011). On the 20 th
century plant become the emerging medicinal products that have been discovered using
extract and some modern medicines comes from pure compounds isolated from plants.
Ethno medicine helps researcher to conduct new study with basis of this traditional plant
(Bernhoft, 2008).
The medicinal effect of the plants that is used as ethno medicines or for the
development of new medicine is the reason that plant produces secondary metabolites
that are no longer needed but these metabolites are the ones that give effect for medicinal
use (Kala, 2000& Parr, et al, 2000). Developed countries are using this secondary by
isolating the compound to create a more potent and effective medicine however,
developing countries like our country opted for herbal medicines, due to the lack of
facilities and fund to able isolate pure compounds (Hudzicki, 2009).
The medicinal benefits of this herb are dated in all the early records of the human
existence. It started during the early Chinese era, which is dated more than 4000 years
ago up to our generation today, the extracts of these herbs and plants are still being used
to treat diseases today. Medicinal herbs are being used extensively in our generation and
are being expanded into much noncomplex forms. This study is then focused on the
synthesize as an antioxidant, antimicrobial, and many more. Other studies found out
those flavonoids are just not antioxidants but also exhibits antiallergenic, antiviral, antiinflammatory and vasodilation actions (Pietta, 2000).
However, to test the antimicrobial activity Paspalum conjugatum and is to
conduct an in vitro study by which testing it through susceptibility of common bacteria
that are commonly the causes of infections such as Staphylococcus aureus (Uwaezuoke,
& Aririatu, 2005). Through Kirby-Bauer agar diffusion method,the in vitro study is used
to test the susceptibility of bacteria against the extracts of PaspalumConjugatum, which
is efficient and reliable in screening compound of natural extract of the plant (Sarker, et
al, 2007).
It has been alarming that more and more of bacteria are resistant to drugs
including Staphylococcus aureus (Locksley, et al, 1982). As antibacterial becomes to
falter from its efficacy, it also has never ending complaints of the side effects that brought
together upon taken (Uwaezuoke, &Aririatu, 2005). It is also the same in the locality but
rather the inevitable growing population and poverty, that drags health services to
implacably unavailable to those who needed it most (Chan, 2010 & WHO, 2011).
With the majority found on common damp environments, Pseudomonas
aeruginosa is a rod-shaped, strict aerobe. Furthermore, it is regularly found in soilbut
more in waterborne creatures. However, colonization has been found in numerous
different plants and organisms, thus it appeared to bring a colony of more than fifty
percent of the people.
Pseudomonas aeruginosa will be an entrepreneurial pathogen that camwood
actuate infections that regularly bring about hospitalization and are frequently lives
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through contact with creatures or persons (Center for Disease Control and Prevention,
2014).
E. coli comprises of a various gathering of microbes. Pathogenic E. coli strains
are sorted into pathotypes. Six pathotypes are connected with looseness of the bowels and
all things considered are alluded to as diarrhea genicE. coli.
Synthesis
Treatment of infectious diseases in this millennium shows significant side effects.
Too much intake of this medicine might cause the bacteria to produce resistance to this
drug, thusthis dilemma is inescapable. To help and minimize this problem, an in vitro
study was conducted that would determine the susceptibility of bacteria against the
extract of Paspalum conjugatum.
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CHAPTER 3
METHODOLOGY
Research Design
The study was carried out through in vitro study where a strict and proper
observation is required in conducting the antimicrobial assay and keeping it from
contamination wherein samples are cultured bacteria and isolated in a prepared agar, it
included two types of bacteria. Two from Gram-positive bacteria (Staphylococcus aureus
and Pseudomonas aeruginosa) and one from Gram negative bacteria (Escherichia coli).
The bacteria were cultured and obtained in the microbiology laboratory of the
Department of Biological Sciences. Kirby-Bauer zone of inhibition test was used in
determining antimicrobial activity of the decoction extract.
In the Kirby-Bauer antimicrobial test, microbial suspension containing bacteria
were spread evenly onto the surface of the solidified medium. The plates were allowed to
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solidify for 15 minutes before the test discs were placed at equidistance from each other.
The top cover of plates was marked as each filter disc paper was placed to create zones of
inhibition. The petri dish of culture group bacteria consists of four filter paper, two (2) for
the plant extract and two (2) for standard group. The assay was left for 18 to 24 hours.
For the zones of inhibition, Vernier caliper was used in measuring.
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Fresh plant parts were collected near the Andrada heights subdivision, Iligan
City.Fresh plant materials were washed and cleaned thoroughly under running tap water,
and then brought to the College of Nursing (CON) of Mindanao State University Iligan
Institute of Technology (MSU-IIT)afterdecoction.
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the negative and positive group and two other filter paper discs impregnated with various
extracts.
After the assay procedure is completed, the Petri dishes were incubated in an
inverted position at 37C for 18 to 24 hours for bacteria to allow growing. The following
day, observation was made.
Zones of Inhibition
The diameters of the zone of inhibition (ZOI), defined by the clear area devoid of
growth, were measured twice. The anti-microbial activities were determined by the ratio
of the ZOI diameters of the extracts to that standard antibiotic is in the same petri dish.
This was measured through Vernier caliper of the distance on the inhibition from filter
paper disc.
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CHAPTER 4
RESULTS AND DISCUSSION
Table 1 displays the results of the antimicrobial assay done with the Carabao
Grass decoction on the four types of organisms namely: Bacillus subtilis, Escherichia
coli, Pseudomonas aeruginosaand Staphylococcus aureus.
The samples given were tested against a positive control, which is
chloramphenicol, a broad-spectrum antibiotic drug, and a negative control, which is
sterile water. The results showed that Carabao Grass decoction has no antimicrobial
activity on all of the test organisms employed in the study. The samples given did not
yield Zones of Inhibition (ZOI).
Table 1 Average Zones of Inhibition (ZOI) in millimeters (mm)
R1
R2
R3
Negative
control
(Distilled
Water)
Bacillus subtilis
22.24
Staphylococcus aureus
17.08
Escherichia coli
12.00
Pseudomonas aeruginosa
4.25
Decoction Extract
Test Organism
Positive control
(Chloramphenic
ol)
Based on the result, the table showed no antimicrobial activity of the Carabao
decoction extract. It yielded no zones of inhibition (ZOI) from replicates one, two, and
three.
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The Carabao grass contains chemical compounds such as the flavonoids that are
capable of killing microorganisms. However, the data is in contrary to the study of
Antimicrobial Activity of Betula pendula ,in which the decoction of buds and flowers of
the plant yielded positive inhibition in the Zone of Inhibition with the same bacteria
sample.(Duric et.al., 2013) in which it has antimicrobial effects. The distilled water on
the other hand as negative control yielded an expected zeros result because it does not
contain any antimicrobial properties. The positive control, which is the Chloramphenicol,
yielded antimicrobial activity against the four test organisms such as Bacillus subtilis
(22.24), Staphylococcus aureus (17.08), Escherichia coli (12.00), and Pseudomonas
aeruginosa (4.25 mm).The positive control is a broad-spectrum antibiotic that is
commonly used in the hospital due to its known killing effects on microbes, availability
in the pharmacy, and affordability. In comparison, the decoction extract is no match to the
positive control.
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CHAPTER 5
SUMMARY OF FINDINGS, CONCLUSIONS AND RECOMMENDATIONS
Summary
The study of the Antimicrobial Activity of Carabao Grass (Paspalumconjugatum)
is a quantitative experimental research. It was done to gather data and facts in the
management of various infections and at the same time a cost-saving intervention to
people in the community that are not financially stable.
The antimicrobial potency of Carabao grass decoction extract was assessed
through the Kirby-Bauer Disc Diffusion Test.
The study was conducted within the period of January to May 2016 in Iligan City,
specifically at MSU- IIT College of Nursing and the Department of Biological Sciences
(DBS) in the College of Science and Mathematics (CSM). As to sample, plants were
gathered in the local area of Iligan City, cleansed from roots to leaves before subject to
decoction. Upon obtaining the sample, extract of Carabao Grass were then forwarded to
laboratory for antimicrobial assay. The samples given were tested against a
chloramphenicol, a broad-spectrum antibiotic as a positive control and distilled water as
negative control. Results of the antimicrobial assay were collected after 24 hours of
determining Zones of Inhibition (ZOI).
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Conclusion
The results showed that the Carabao grass (Paspalum conjugatum) had no
antimicrobial activity in all three replicates of the four test microorganisms such as the
Bacillus subtilis, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa.
The decoction extract was of no match against the broad-spectrum antibiotic, which is the
Chloramphenicol that yielded antimicrobial activity. In this finding, it could be the said
herbal plant usually used and practiced in the community were not that efficient in
deterring the microbes as used for wound healing. Despite the findings, it was not
conclusive to say that all Carabao grass decoction extract has no effect on killing
microorganisms. Further investigation should be done before we can conclude that the
said sample is not potent for antimicrobial effect.
Recommendation
The following recommendations are drawn on this study based on the results:
1. To conduct a more extensive research and experimentation for Carabao Grass
(Paspalum conjugatum) not only for antimicrobial testing but also for the
antifungal testing and phytochemical analysis.
2. To obtain bigger sample of the Carabo grassfor a better antimicrobial activity . We
also recommend the future researchers to reconduct the antimicrobial testing and
if the result will still be zero they should inform and conduct an awareness
program to the community that this herb or grass is not a potent herb to be used as
an alternative antimicrobial agent to treat their diseases.
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BIBLIOGRAPHY
BOOKS
Austin, D. F., FL. (2004).Florida Ethnobotany, pp. 482. Suite 300 Boca Raton, CRC
press. 6000 Broken Sound Parkway, Norway.
Cambie, R.C., Ash, J., (1994). Fijian Medicinal Plants. pp. xvi. CSIRO. Australia.
KALA,CP (2006). Medicinal Plants and Sustainable Development. pp. 43-56. Nova
Science Publishers Inc. New York.
ARTICLE
Perry, L.M., (1980) Medicinal plants of East and South East Asia. 620 pp. MIT Press.
Cambridge. Massachusetts.
JOURNAL
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Multiple
antibiotic
resistant
Staphylococcus
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WEB SOURCE
Pseudomonas
aeruginosa.
Retrieved
from
subtilis.
Retrieved
http://web.mst.edu/~microbio/BIO221_2009/B_subtilis.html
from
May
25,
23
2016Escherichia
coli
retrieved
from
http://www.microbelibrary.org/component/resource/laboratory-
test/3189-kirbybauer- disk-diffusion-susceptibility-test-protocol
Carabao Grass for Kidney Disease. (2014, December 20). Retrieved from
http://www.pkdclinic.org/chinese-herbs/2140.html
Burkill, H.M. (1985). The useful plants of west tropical Africa, Vol 2. Retrieved from
http://plants.jstor.org/stable/10.5555/al.ap.upwta.2_659
Russo, E. B., Headache Treatments by Native People of the Ecuadorian Amazon: A
Preliminary
Cross-Disciplinary Assessment. Retrieved from
http://manu.montana.com/paper.html
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APPENDIX
A
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Mindanao State
University
Iligan Institute of Technology
College of Nursing
Dear Sir,
Good Day!
The students listed below from MSU-IIT College of Nursing will be
conducting an experimental research in partial fulfilment of the requirements
for the degree of Bachelor of Science in Nursing.
In line with this, we would like to request from your department to
authenticate the provided herbal plant to be used of the said experiment.
This will help us fulfil our Nursing Research and hopefully present a research
paper on the same school year.
Hoping for your kind consideration and approval. More power and God
Bless!
Truly yours,
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APPENDIX
B
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