ANTIMICROBIAL ACTIVITY OF GINGER (Zingiber officinale) AND
CALAMANSI (Citrus microcarpa) PEELS EXTRACTS AGAINST WOUNDINFECTING BACTERIA
A Research Paper
Presented to the Faculty of the
Senior High School
Science, Technology, Engineering, and Mathematics
Zamboanga del Norte National High School
Turno Campus, Dipolog City
In Partial Fulfillment of the Requirements for
Inquiries, Investigation, Immersion I
Experimental Research
NIÑO NEIL P. TAPIZ
DHINA ELIJAH LANE D. JAIME
AARON JHAN C. POTOY
JV FRANC CHRIS A. ABAD
NATHAN NYLE E. EVARDO
MARK JOSHUA E. LORION
MARY JOY B. JANOLINO
JOHN PAUL A. ACOPIADO
February 2023
ABSTRACT
Tapiz, N. N. P., Jaime, D. E. L. D., Lorion, M. J. E., Janolino, M. J. B.,
Potoy, A. J. C., Abad, J. F. C. A., Evardo, N. N. E. E., & Acopiado, J. P. A.
(2023). Antimicrobial Activity of Ginger (Zingiber officinale) and Calamansi
(Citrus microcarpa) Against Wound-Infecting Bacteria. Unpublished Research
Paper. Science, Technology, Engineering, and Mathematics. Senior High
School. Zamboanga del Norte National High School. Dipolog City
Due to the expensive medicines in the Philippines, the people can’t access
simple medications such as antimicrobial creams. This sparks the researchers to have a
deeper scientific research on alternative medicines. Medicinal plants have managed
several diseases caused by infections. Ginger peels possess both antioxidative and
antimicrobial properties and calamansi can be used to make antimicrobial juice that can
be used to treat pathogenic bacteria. Meanwhile, this study sought to investigate the
antimicrobial activity of calamansi peels extracts, ginger peels extracts, and calamansi
and ginger peels extracts against wound infecting bacteria. Parallel Group Design was
utilized throughout all of the experiments to compare the experimental groups.
Wherein, in investigating the antimicrobial activity of ginger and calamansi peels
extracts against wound infecting bacteria, there were 3 treatment groups which would
receive ginger extracts, calamansi extracts, and combination of ginger and calamansi
extracts, respectively. Zone of inhibition is used to gather the data. As a result,
calamansi peels extract has a mean of 0.8mm which is prescribed as “+” <15mm,
meaning it has a small effect. Meanwhile ginger peels extract has a mean of 0 mm
which is prescribed as “-” no zone, and the combination of calamansi and ginger peels
extracts also prescribed as “-” no zone for attaining 0 mm in zone of inhibition.
Afterwards, there was no significant difference between the variables after having a pvalue of 0.110, greater than the significant at 0.05 level as tested using One-Way
ANOVA. This study concludes based on its methodology that calamansi peels extracts
have observable antimicrobial activity against wound-infecting bacteria compared to
ginger peels extracts. Also, this study suggests using other medicinal plants and
exploring other extraction methods.
Keywords: calamansi, ginger, antimicrobial, wound-infecting bacteria, zone of
inhibition, parallel group design, extracts
Chapter I
THE PROBLEM AND ITS SCOPE
Introduction
In recent times, cases of occupational injuries and occupational accidents
decreased from 2017 to 2019. In 2019, 37,513 occupational accidents were reported
across all industries in the Philippines. Work-related accidents occurred in around 10.9
percent (4,186) of the total 38,305 establishments employing 20 or more workers
(Philippine Statistics Authority, 2022).The majority of those cases were open wounds
that could be applied with first aid treatment to avoid being infected as much as
possible.
Applying first aid treatment greatly helps the patient. First aid is immediate
medical attention given to an injured individual. The goal of first aid is to prevent
damage and eventual incapacity. In extreme instances, first assistance may be required
to keep the victim alive. These first aid measures should be available in every institution
and provided by the employer of the company. According to the Labor Code Law of
the Philippines, Article 156 is entitled "First Aid Treatment." In accordance with
regulations prescribed by the Department of Labor and Employment, every employer
is required to keep in their establishment the necessary first-aid medicines and
equipment that may be required based on the nature and conditions of the work.
First aid kits typically include over-the-counter medications and antiseptic
creams. However, in the Philippines, the high cost of medicines makes it difficult for
people to access these goods. A survey conducted by Pulse Asia in 2019, cited by the
Department of Health in the same year, revealed that 99% of Filipinos choose not to
purchase medications due to the high prices.
The high cost of medications in the Philippines has sparked researchers' interest
in conducting deeper scientific research on alternative medicines. Medicinal plants, for
example, had been found effective in treating various infectious diseases. Although the
specific causal variables within plant-based medicine are not well understood, the
success of a herbal medicine in treating infections highlights the significant effects of
numerous plants on bacterial, fungal, or parasitic infections (lonescu, 2017).
The antimicrobial properties of a certain plant wastes were utilized to create lowcost antiseptic topical medications. A study conducted by Afolayan A.J., Otang, W.M.
(2015) demonstrated the high antimicrobial properties of citrus lemon peels extract.
Additionally, the study conducted by Mbaeyi-Nwaoha, et al. (2013) concluded that
ginger peels possess both antioxidative and antimicrobial properties, indicating that no
part of the ginger plant can be considered waste. Cubias, J.G., et al., (2014) conducted
a study titled "Preparation of an antimicrobial extract from seeds, pulp, and peelings of
Philippine calamansi (Citrus mitis) fruit against E. coli (Escherichia coli)," which
showcased the extraction of antimicrobial juice from calamansi that can be utilized
against pathogenic bacteria.
Antiseptics were frequently employed in medical care to eliminate or halt the
growth of microorganisms on the skin and mucous membranes. They were also utilized
for cleansing hands and treating minor injuries. According to Robert(2022), antiseptics
were chemical agents that were applied to the skin to reduce the microbial count and
minimize the risk of infections.
Similar to earlier investigations, this one was concerned with figuring out which
variables have antibacterial properties. By examining the antibacterial activities of
ginger (Zingiber officinale) and calamansi (Citrus microcarpa) peel extracts, it sought
to fill in the knowledge gaps in those earlier studies.
Conceptual Framework
This study was anchored to research studies related to the antimicrobial
activities of ginger (Zingiber officinale) and Calamansi (Citrus microcarpa) calamansi.
The study of Shivakumar N, et al.,(2020) entitled “Ginger Cultivation and Its
Antimicrobial and Pharmacological Potentials” shows that ginger extracts had
antimicrobial activity against a wide spectrum of pathogenic organisms. Meanwhile, a
study conducted by Cubias, J.G., et al., (2014) entitled, Preparation of an antimicrobial
extract from seeds, pulp and peelings of Philippine calamansi (Citrus mitis) fruit against
E.coli (escherichia coli) shows that calamansi can be extracted to produce antimicrobial
juice, and can be used against pathogenic bacteria.
This study was similar to previous studies that focused on determining the
antimicrobial activities of certain variables. However, its objective was to address the
research gaps in those previous studies by exploring the antimicrobial properties of
ginger (Zingiber officinale) and Calamansi (Citrus microcarpa) peel extracts.
Figure 1 depicted the study's schema. The box located in the lower left portion
of the figure was labeled as the input, representing the independent variables, namely,
calamansi peels and ginger peels. The lower middle box of the figure was labeled as
the process, representing the intervening variable, which involved the extraction
process of calamansi and ginger peels. Finally, the box in the lower rightmost part of
the figure was labeled as the output, indicating the dependent variables, which were the
results of the antimicrobial activity of the extracts against wound infecting bacteria.
Input
Process
Output
•Calamansi
Peels
•Ginger Peels
Calamansi and
Ginger Peels
Extraction
Antimicrobial
Activity against
Wound-Infecting
Bacteria
Figure 1.1. Antimicrobial properties of calamansi (Citrus microcarpa) and ginger
(Zingiber officinale) against wound-infecting bacteria
Objective of the Study
The main objective of this study was to determine the antimicrobial activity of
extracts from ginger (Zingiber officinale) and calamansi (Citrus microcarpa) against
bacteria that causes wound infections. Additionally, the study aimed to formulate an
antiseptic cream using these extracts and evaluate its effectiveness in wound healing on
albino mice, comparing it to commercially available antiseptic topical creams. The
findings of this study would provide valuable insights for the development of a natural,
safe, and affordable alternative to existing commercial antiseptic creams.
Statement of the Problem
This study has specific aims to address the following questions:
1. What is the antimicrobial activity of ginger extracts in terms of the Zone of
Inhibition?
2. What is the antimicrobial activity of calamansi extracts in terms of the Zone of
Inhibition?
3. What is the antimicrobial activity of ginger and calamansi extracts in terms of
the Zone of Inhibition?
4. What are the significant differences in the antimicrobial properties between
ginger (Zingiber officinale) peel extract, calamansi (Citrus x microcarpa) peel
extract, and the combination of ginger (Zingiber officinale) and calamansi
(Citrus x microcarpa) peel extracts in terms of the Zone of Inhibition?
Hypothesis
Based on the problems mentioned, hypothesis were formulated.
H0: There is no significant difference between the antimicrobial properties of
ginger (Zinger officinale) peels extract, calamansi (citrus x microcarpa) peels extracts,
and combination of ginger (Zinger officinale) and (citrus x microcarpa) peels extracts
in terms of zone of inhibition.
Significance of the Study
This study undertaken by the researchers conveyed valuable information
which was grounded on its objectives. This study, however, was not undertaken without
its beneficiaries in order to become more substantial and useful. Thus, the study was
deemed significant to the following.
Community. This study would help the people in the community find innovative but
affordable ways to produce alternative antiseptic cream as a first aid to open wounds.
Environment. The materials needed are the peels of calamansi and ginger which are
commonly thrown away. With this, agricultural waste would be lessened which can benefit
our environment.
Health Institutions. This study will be significant to the health institution to procure new
methods or medicine in dealing with wound-infecting bacteria. Also, health institutions can
produce low-cost antiseptic creams that are effective in treating wound infections. This can
be particularly beneficial in low-cost income areas where people may not be able to afford
expensive medications.
Other Researchers.
This study covers information involving the medication as an
approach to healing wounds. Thus, the result of this study can be used for future discussions
on the capabilities of medication for other types of wound concerns.
Scope and Delimitations
The study aimed to investigate the antimicrobial activity of ginger and
calamansi peel extracts against wound-infecting bacteria. It involved three
experimental groups: ginger peel extracts, calamansi peel extracts, and a combination
of calamansi and ginger peel extracts. The purpose was to measure the effectiveness of
the extracts in inhibiting bacterial growth.
In the study, the researchers aimed to evaluate the inhibitory effects of the three
experimental groups on wound-infecting bacteria. They prepared their own bacterial
agar growth medium to culture the bacteria and applied their extracts to measure the
diameter of the zone of inhibition for each of the three treatment groups. It's important
to note that this experiment focused solely on investigating the antimicrobial activity
of ginger and calamansi peel extracts against wound-infecting bacteria. The researchers
did not conduct further studies on the bacteria present in the wound, nor did they test
specific bacteria from wounds.
The study was conducted at Zamboanga del Norte National High School, Turno
Campus, and at the house of one of the researchers. It took place between March 2023
and May 2023.
Definition of Terms
To provide a clearer and better understanding of the terms used in the study,
the following are defined:
Agar. A gelatinous material derived from algae, specifically used as a culture medium
of bacteria and other cells for diagnostic or laboratory purposes. In this study it is used
to test the microbial activity of the ginger and calamansi extract through a zone of
inhibition.
Agar Solution. A hydrophilic colloid extracted from certain seaweeds of the
rhodophyceae class. It is insoluble in cold water but soluble in boiling water.
Antiseptic. Relating to or denoting substances that prevent the growth of disease
causing microorganism. In this study, antiseptic refers to the ability of the low-cost
topical medication cream to prevent the growth of microorganisms and disinfect the
wound, thus reducing the risk of infection.
Antimicrobial Activity. A collective term for all active principles (agents) that inhibit
the growth of bacteria, prevent the formation of microbial colonies, and may destroy
microorganisms. In this study, antimicrobial activity refers to the ability of the extracts
obtained from ginger and calamansi peels to inhibit the growth or kill wound-infecting
bacteria.
Bacteria Solution. A liquid preparation called a bacterial solution contains bacteria
suspended in a medium or solvent. It is frequently employed in scientific studies,
academic research, and commercial applications.
Calamansi (Citrus microcarpa). A citrus fruit that is commonly used in Southeast
Asian cuisine and traditional medicine for its antimicrobial and anti-inflammatory
properties. In this study, the peel of the Citrus microcarpa fruit is used to extract
bioactive compounds with potential antimicrobial properties.
Ethyl Alcohol (Ethanol). An organic compound, it is a volatile, flammable, colorless
liquid with a characteristic wine-like odor and pungent taste. In this study, ethanol is
used for the extraction of calamansi and ginger peels.
Ethanolic Extraction. A method of employing ethanol (ethyl alcohol) as a solvent to
extract bioactive chemicals or compounds from plant material. It is a commonly utilized
technique in many different industries, including food processing, herbal medicine, and
pharmaceuticals.
Kirby Bauer Diffusion Susceptibility Protocol Test. A method used to get the zone
of inhibitions of the extracts. In this study, disks with known concentrations of an
antimicrobial agent are placed on the surface of an agar plate containing a
nonselective medium that has been inoculated with a suspension of a strain to conduct
the disk-diffusion susceptibility test.
Peels Extracts. Refers to the active compounds that are extracted from the outer layer
of ginger and calamansi fruits. In this study, extracts refers to the concentrated form of
bioactive compounds obtained from ginger and calamansi peels by using solvents such
as water or alcohol.
Ginger (Zingiber officinale). A flowering plant that is widely used as a spice and in
traditional medicine for its anti-inflammatory and antioxidant properties. In this study,
the rhizome or root of the ginger plant is used to extract bioactive compounds with
potential antimicrobial properties.
Wound-Infecting Bacteria. Refers to bacteria that cause infections in wounds, such
as Staphylococcus aureus and Pseudomonas Aeruginosa.
Zone of Inhibition. Used to determine the susceptibility or resistance of pathogenic
bacteria to antibacterial agents.
Chapter II
REVIEW OF RELATED LITERATURE AND STUDIES
This chapter discusses foreign and local journals, articles, and studies associated
with the present research study.
Related Literature
Ginger (Zingiber officinale)
Ginger is a spice that has been used for medicinal purposes for thousands of
years. It has several health benefits, including anti-inflammatory, antioxidant, and
antimicrobial properties. One of the most well-known health benefits of ginger is its
antiseptic properties.
Several studies have shown that ginger has strong antimicrobial activity against
a variety of bacteria, viruses, and fungi. The present study showed the potent
antimicrobial activity of the ginger extract against the all tested bacterial pathogens,
Islam (2014). In the study of Xin et al.(2020) Ginger is used as a spice in foods and
beverages because of its characteristic spicy aroma and taste. In addition, it is an
excellent source of many bioactive compounds, including bioactive phenols (gingerols,
shogaols, and zingerones). Ginger essential oil (GEO) is the volatile oil extracted from
the root of ginger. Due to its unique fragrance and biological activity, it has a very broad
development prospects in the pharmaceutical, food, and cosmetics industries
Another study of Jundishapur et al,.(2016) found that ginger extract was
effective against several types of fungi, including Candida albicans, which is a
common cause of yeast infections. According to the same study above, it has been
demonstrated that ginger extract has potent antifungal properties against fluconazoleresistant Candida albicans species isolated from patients with genital candidiasis.
Indeed, in traditional medicine, ginger is administered to cure movement inabilities,
nausea, and vomiting during pregnancy. More importantly, apart from sedation and
drowsiness, there is no report of any side effects for ginger
Furthermore, a study of Jung San et al,.(2013) found that ginger extract had
antiviral activity against the respiratory syncytial virus (RSV), which is a common the
study of Shrestha et al., (2020) conducted a study in a tertiary care hospital in Nepal to
investigate the antibiotic resistance patterns of bacterial strains isolated from wound
infections. They found that the bacterial strains were highly resistant to commonly used
antibiotics, highlighting the urgent need for alternative treatments.
A study by Abdullahi et al., (2019) reviewed the literature on wound infections
and antimicrobial resistance in small animal practice. They found that the emergence
of antimicrobial-resistant bacterial strains has become a major challenge in the
treatment of wound infections in small animals. The authors recommend the
development of alternative antimicrobial strategies to address this issue. In the
concluding study of Puca et al., (2019) also investigated antibiotic resistance in
bacterial pathogens and its potential implications in the treatment of infected wounds.
They discuss the various factors contributing to the emergence of antibiotic resistance,
including the overuse and misuse of antibiotics. The authors suggest that the
development of new antimicrobial agents and the implementation of effective infection
control measures are crucial in combating antibiotic resistance.
These related studies highlight the growing concern of antibiotic resistance in
wound infection bacteria and the urgent need for alternative treatments and strategies
to address this issue as a cause of respiratory infections. Jung San et al,.(2013) Fresh,
but not dried, ginger is effective against HRSV-induced plaque formation on airway
epithelium by blocking viral attachment and internalization.
Calamansi (Citrus mircrocarpa)
One study by Trakranrungsie and Chatchawanchonteera et al., (2016)
investigated the antimicrobial activity of calamansi peel extract obtained through
ultrasound-assisted extraction against several foodborne pathogens. The study found
that the extract exhibited significant antimicrobial activity against all tested bacteria,
suggesting its potential for use as a natural preservative in food and as an antiseptic
agent in wound care.
In the study of Jayaweera et al., (2019), a combination of ginger and calamansi
peel extracts obtained through supercritical fluid extraction was used for the
development of a low-cost antiseptic cream for wound care. The study found that the
cream exhibited significant antimicrobial activity against wound-infecting bacteria
such as Staphylococcus aureus, suggesting the potential of calamansi peel extract for
use in low-cost wound care products. A study by Giron et al., (2017) investigated the
antibacterial activity of calamansi peel extract obtained through maceration against
Staphylococcus aureus. The study found that the extract exhibited significant
antibacterial activity, which was attributed to the presence of various bioactive
compounds such as flavonoids, alkaloids, and tannins.
These literatures suggested that calamansi peel extracts had significant potential
for use as natural antimicrobial agents in wound care. Further research is needed to
optimize the extraction methods and formulations of these extracts for effective use in
wound care products.
Plant Extracts/Extraction
According to Abdullahi, et al. (2022), the most important sources of
biomolecules that can be screened from plant components are thought to be plant
extracts. These biomolecules can be extracted from medicinal plants using a variety of
solvents and extraction techniques. The former focused on the antimicrobial properties
of plant extracts and their potential as natural alternatives to synthetic antimicrobials,
while the latter provides an overview of the traditional use of medicinal plant extracts
in healthcare systems around the world.
The use of plant extracts as antimicrobials is becoming increasingly popular due
to the rise of antibiotic-resistant bacteria. It emphasizes the potential of plant extracts
as a new generation of antimicrobials that could help combat this issue. It shows various
plant extracts that have been shown to possess antimicrobial activity against a range of
pathogens. It also highlights the advantages of using plant extracts over synthetic
antimicrobials, such as lower toxicity, fewer side effects, and greater availability.
In the study by Trakranrungsie and Chatchawanchonteera (2016), they
evaluated the antimicrobial activity of calamansi peel extract obtained through
ultrasound-assisted extraction against several foodborne pathogens. The study found
that the extract exhibited significant antimicrobial activity against all tested bacteria,
suggested its potential for use as a natural preservative in food and as an antiseptic agent
in wound care. A study by Jayaweera et al. (2019) The study utilized extracts from
ginger and calamansi as the main active ingredients due to their well-known
antimicrobial activity. The results indicated that the cream had significant antimicrobial
activity against wound-infecting bacteria.
Antimicrobial
The study of Akinmoladun, et al.,(2018) investigated the antibacterial properties
of ethanolic extract of ginger rhizome and Citrus microcarpa (Calamondin) peel
extracts and the synergistic activity of ginger and lemon extracts against methicillinresistant Staphylococcus aureus (MRSA). The ethanolic extract of ginger rhizome
showed significant antibacterial activity against several wound-infecting bacteria,
including S. aureus, Pseudomonas aeruginosa, and Escherichia coli. The extract also
exhibited potent antioxidant and antiproliferative activities.
The Citrus microcarpa (Calamondin) peel extracts showed antibacterial activity
against various foodborne pathogens, including S. aureus, E. coli, Salmonella typhi,
and Listeria monocytogenes. According to the study Zahoor, et al.,(2020)
Phytochemical analysis of the extract revealed the presence of several active
compounds, including flavonoids, tannins, and phenols. In the study of Ayuob et al.,
(2013) synergistic activity of ginger and lemon extracts against MRSA was
investigated, and the results showed that the combination of both extracts had a
significant inhibitory effect against MRSA compared to the individual extracts. The
study suggests that combining ginger and lemon extracts may enhance the antimicrobial
activity against MRSA, a notorious bacterium known for its resistance to various
antibiotics.
This literature provided evidence of the potential of natural products such as
ginger and citrus extracts as sources of antimicrobial agents that could be useful in
developing low-cost antiseptic topical medication creams for the treatment of wound
infections.
Wound-Infecting Bacteria
Infections that develop in a cut or other skin wound are called wound infections.
Many microorganisms, such as Staphylococcus aureus, Streptococcus pyogenes, and
Pseudomonas aeruginosa, can result in wound infections. Through a skin break, these
germs can enter the body and lead to an infection. It's critical to prevent wound
infections in order to avoid problems like abscesses, cellulitis, or even sepsis.
Related Studies
Ginger Antimicrobial Activity
A study by Koffi-Nevry et al. (2019) evaluated the in vitro antimicrobial activity
of ginger and garlic extracts against bacterial strains isolated from wound infections.
The results showed that both extracts exhibited significant antimicrobial activity against
the tested bacteria, suggesting their potential as natural antimicrobial agents for wound
healing. The study of Chakraborty and Ghosh (2020) reviewed the antimicrobial
activity of ginger extract against foodborne pathogens and its potential application in
food systems. The review summarized several studies that demonstrated the
antimicrobial properties of ginger extract against various foodborne pathogens,
including Escherichia coli, Salmonella, and Listeria monocytogenes.
The study conducted by Osho et al. (2020) investigated the antibacterial and
phytochemical properties of ginger extract against selected foodborne pathogens. The
results showed that ginger extract had significant antibacterial activity against the tested
bacteria, including Staphylococcus aureus, Escherichia coli, and Salmonella enterica.
The study also identified several bioactive compounds in the ginger extract that were
responsible for its antibacterial activity. A study conducted by Ghasemi and Heydari
(2020) reviewed the potential of ginger and its active compounds as candidates for the
prevention and treatment of infections caused by Gram-positive bacteria. The review
discussed the antimicrobial activity of ginger extract and its bioactive compounds
against different types of Gram-positive bacteria, including Staphylococcus aureus,
Streptococcus mutans, and Enterococcus faecalis.
These related studies suggest that ginger extract and its bioactive compounds
have significant antimicrobial activity against various types of bacteria, including those
that cause wound infections and foodborne illnesses. Therefore, they have potential
applications in the development of natural antimicrobial agents for various purposes.
Calamansi Antimicrobial Activity
In the study of Villaflores et al. (2017) evaluated the antimicrobial activity of
calamansi fruit extracts and essential oils against foodborne pathogens, and found that
the extracts exhibited inhibitory effects against all tested pathogens. The Study of
Ongsakul et al. (2019) investigated the antimicrobial activity of Citrus microcarpa fruit
extracts against some human pathogenic bacteria, and found that the extracts had the
potential to be used as natural antimicrobial agents.
In the Study of Mohammadifard et al. (2021) studied the antibacterial activity
of calamansi peel extracts against foodborne pathogens and found that the extracts
exhibited significant antibacterial activity, with potential use as a natural alternative to
synthetic antimicrobial agents. Chantarasataporn et al. (2020) evaluated the
antimicrobial and antioxidant activities of calamansi peel extracts and found that the
extracts exhibited potent antimicrobial and antioxidant activities, suggested that they
could be used as natural preservatives and antioxidants in food products.
These related studies suggest that calamansi extracts may have potential as a
natural alternative to synthetic antimicrobial agents in the development of low-cost
antiseptic topical medication cream, especially when combined with other natural
extracts such as ginger.
Ginger and Calamansi Antimicrobial Activity
A study of Karuppiah et al,.(2012) investigated the antimicrobial activity of
ginger against a range of bacteria, including Escherichia coli, Staphylococcus aureus,
and Bacillus subtilis. The researchers found that ginger extract exhibited significant
antibacterial activity against all the tested bacteria, and concluded that ginger could be
a useful natural antimicrobial agent.
Another study published by Bascon et al., (2020) evaluated the antimicrobial
activity of calamansi against a range of pathogenic bacteria, including Salmonella
typhimurium, Staphylococcus aureus, and Escherichia coli. The researchers found that
calamansi extract exhibited strong antimicrobial activity against all the tested bacteria,
and suggested that calamansi could be a potential source of natural antimicrobial agents.
Wang (2020) investigated the synergistic antimicrobial activity of ginger and
calamansi against Escherichia coli and Staphylococcus aureus. The researchers found
that a combination of ginger and calamansi extracts exhibited stronger antimicrobial
activity against the bacteria than either extract alone, and concluded that the two plants
could be used together as natural antimicrobial agents in food preservation.
A study published by FBFS (2021) examined the antimicrobial activity of
ginger and extracts against various foodborne pathogens, including Listeria
monocytogenes, Salmonella enterica, and Escherichia coli. The researchers found that
ginger extracts exhibited significant antimicrobial activity against the tested pathogens,
and suggested that they could be used as natural preservatives in food products.
Overall, these studies suggest that both ginger and calamansi have potent
antimicrobial properties and could be used as natural alternatives to synthetic
antimicrobial agents in food preservation and other applications. However, more
research was needed to fully understand the mechanisms underlying their antimicrobial
activity and to determine their efficacy and safety in different contexts.
Wound-Infecting Bacteria Resistance
In the study of Shrestha et al., (2020) conducted a study in a tertiary care hospital
in Nepal to investigate the antibiotic resistance patterns of bacterial strains isolated from
wound infections. They found that the bacterial strains were highly resistant to
commonly used antibiotics, highlighting the urgent need for alternative treatments.
A study by Pires et al., (2019) reviewed the literature on wound infections and
antimicrobial resistance in small animal practice. They found that the emergence of
antimicrobial-resistant bacterial strains has become a major challenge in the treatment
of wound infections in small animals. The authors recommend the development of
alternative antimicrobial strategies to address this issue. In the concluding study of
Jneid et al., (2019) also investigated antibiotic resistance in bacterial pathogens and its
potential implications in the treatment of infected wounds. They discuss the various
factors that contributed to the emergence of antibiotic resistance, including the overuse
and misuse of antibiotics. The authors suggest that the development of new
antimicrobial agents and the implementation of effective infection control measures are
crucial in combating antibiotic resistance.
These related studies highlight the growing concern of antibiotic resistance in
wound infection bacteria and the urgent need for alternative treatments and strategies
to address this issue.
Synthesis
These related studies and literature suggest that calamansi and ginger peels have
high antibacterial and antimicrobial potency against different strains of bacteria. Instead
of relying only on commercially made antimicrobial extracts, it acted as an avenue to
look for materials sourced locally that have antimicrobial activities. Wherein extracts
coming from these variables can be used for formulation of low cost antiseptic cream,
especially that demand on low cost antiseptic cream rises.
Chapter III
METHODOLOGY
This chapter presents the sequential methods employed in conducting the
research study and encompasses the research design, necessary materials and
equipment, data gathering procedures, and statistical treatments utilized in the study.
Research Design
The study used a Parallel Group throughout all of the experiments to compare
the experimental groups. In investigating the microbial activity of ginger and calamansi
peels extracts against wound-infecting bacteria, three treatment groups received ginger
extracts, calamansi extracts, and a combination of ginger and calamansi extracts,
respectively. The antimicrobial activity of ginger and calamansi peels extracts in
inhibiting wound-infecting bacteria, in terms of zone of inhibitions, was assessed.
Materials and Equipment
Laboratory Apparatus
(20) 90 mm petri dish
(2) 100 mL Graduated Cylinder
(4) Dropper
(2) 50 mL Graduated Cylinder
(2) forceps
(3) 1 liter beaker
(3) stirring rod
(2) clippers
(5) 500 mL beaker
(2) dissecting kit
Digital Weighing Scale
Vials
Inoculating Rod (L shape)
Inoculating Rod (Triangle)
Equipment
Blender
Incubator
Refrigerator
double boiler
Bullet Blender
Chemicals
Ethanol
Nutrient Agar Solution
Distilled Water
Others
Ginger (Zingiber officinale) peels
Sterile Cotton Swabs
Calamansi (Citrus microcarpa) peels
Aluminum Foil
Sterile Gloves
Details of the Procedure
A. Collection of Materials
The researchers purchased calamansi and ginger from the local market, along
with ethanol and aluminum foil. Additionally, nutrient agar powder, sterile droppers,
sterile cotton swabs, glass vials, and surgical needles were obtained at the local store.
For the remaining materials and apparatus, the researchers borrowed them from
the DOST Chemistry Laboratory, the Senior High School Laboratory at Zamboanga
del Norte National High School - Turno Campus, and their personal equipment.
B. Preparation of Calamansi and Ginger Peels
The researchers planned to purchase 2 kilograms each of ginger and calamansi.
The ginger and calamansi were washed first with tap water and then with distilled water
to ensure cleanliness. Subsequently, the gingers and calamansi were peeled, and their
peels were sun-dried until they reached the desired level of dryness. Once dried, the
peels were further processed into smaller pieces using a bullet blender until they are
reduced to the desired size.
C. Ethanol Extraction of the Calamansi and Ginger Peels
Sixty (60) grams of calamansi peel powder were placed inside a 500 mL beaker
and soaked in 500 ml of 70% ethanol. The beakers were covered with aluminum foil
and left undisturbed in a dry, room temperature area for 24 hours. Similarly, sixty (60)
grams of ginger peel powder were placed in another 500 mL beaker and soaked in 500
ml of 70% ethanol. It was covered with aluminum foil and left in a dry, room
temperature area for 24 hours. Additionally, a mixture of 30 grams of calamansi peel
powder and 30 grams of ginger peel powder were soaked in 500 ml of 70% ethanol
inside a separate 500 mL beaker. This mixture was covered with aluminum foil and
placed in a dry, room temperature area for 24 hours.
Afterwards, the liquid extracts undergo evaporation using the open-dish
evaporation method until they reach a viscous consistency. The viscous extracts from
the three experimental groups were filtered using cheesecloth and transferred into 500
ml reagent bottles. The bottles were stored in a refrigerator for further use.
Furthermore, sterilized round filter papers with a diameter of 6mm will be
soaked in each of the extracts for a period of 24 to 48 hours.
D. Preparation of Agar Growth Solution and Agar Growth Medium
Two (2) liters of distilled water was mixed with 56 grams of nutrient agar. It
was boiled on an electric stove and stirred constantly to ensure that the media was
completely dissolved. Afterwards, the sterilizations were carried out at the Philippine
Red Cross in Dipolog City. At 15 psi and 121° C, the nutrient agar was sterilized in 15
minutes. The other materials were washed with dishwashing soap and water. After
washing, the apparatuses were wiped with a kitchen towel. All laboratory apparatus
used were wrapped with aluminum foil thoroughly and sterilized using an autoclave
with assistance of the Philippine Red Cross - Zamboanga del Norte Chapter laboratory.
It was gently withdrawn from the heat after the heating procedure and was left to cool.
After sterilization, the sterilized nutrient agar solution was distributed into the eight
(20) Petri dishes and kept in the refrigerator to prevent contamination.
E. Obtaining the bacteria
The bacteria were obtained from a wound of a subject being asked by the
researcher through the following accurate research ethics. The researchers swabbed the
wounds using sterile cotton based swabs by gently streaking the wounds. Enough
pressure was used to express the fluids from the wounds. Then the sterile cotton swabs
with wound-infecting bacteria were mixed into a sterile distilled water to produce a
wound-infecting bacteria solution.
F. Applying the Wound-Infecting Bacteria to the Agar Growth Medium
Zero point five (0.2) ml of the wound-infecting bacteria solution was poured
down on each 7 petri dishes, then the spread plate method in inoculation was performed
using an L-shaped inoculating rods. The plates were inverted to avoid any potential
moisture that could interfere with the growth of the bacterial colonies. Then the plates
were incubated at 37° C for 24 hours. The cultured bacteria were checked for microbial
growth after the 24 hours of incubation. The researchers randomly selected successful
plates where microbial growth can be seen to be used for the zone of inhibition of the
extractions.
Using forceps, the filter paper discs which are soaked in each extraction were
placed in the proper location of the petri dishes. Three (3) experimental groups were
made. One (1) experimental group has calamansi peels extract treatment, one (1)
experimental group also has a ginger peels extract, and one (1) experimental group had
the combination of extractions of calamansi and ginger peels. The researchers used
kirby bauer disk diffusion susceptibility test protocol wherein five (5) 6mm
antimicrobial sample disks were placed inside each agar medium while following
proper distance from each antimicrobial sample disks. The petri dishes were labeled
accordingly, they were inverted and incubated at 37° C for 24 hours, and were
rechecked for the next 24 hours incubating after the initial rechecking.
Figure 2 shows the schema of the set-up.
Treatment 1: Calamansi Peels Extract
Treatmet
Treatment 2: Ginger Peels Extract
Treatmet
Treatment 3: Calamansi and Ginger
Peels Extract Treatment
Figure 2. Kirby-Bauer Disk Diffusion Susceptibility Protocol Test placement of
antimicrobial disc sample in a agar medium
G. Data Collection
After the final incubation, a zone of inhibition test was used. They were tested
using the disk diffusion assay against a strain of a common wound bacteria. The
diameters of the inhibition zones were measured using a ruler following the KirbyBauer method. The data in each experimental group was collected and added, and then
the mean of each experimental group was calculated. The diameter was prescribed as
follows: “-” when no antimicrobial effect is noticed, “+” when the zone of inhibition is
<15 mm, and “++” when it is between 15 to 20 mm (Suflet et al., 2021).
Meanwhile, it undergoes a One-Way ANOVA test to determine the significant
difference between the data gathered from the experimental groups.
Statistical Treatment
Mean
The process of calculating the average involves determining the mean value of
a set of collected data. This calculation is performed manually by adding up all the
quantities and then dividing the sum by the total number of quantities. In the context of
the study, to obtain the average diameter of the zone of inhibition for each experimental
group, the arithmetic mean was utilized. This involves calculating the sum of the
diameters of the zones of inhibition and dividing it by the total number of measurements
within each experimental setup.
𝑥=
Where:
∑
𝑥
𝑥i
= sum of all diameters
= number of items
One-Way ANOVA
𝑥𝑥
∑
𝑥
One-Way ANOVA were used to determine if there is a significant difference
between the means of the groups in each variable. Especifically, this statistical tool was
used to determine the significant difference between the antimicrobial properties of
ginger (Zinger officinale) peels extract, calamansi (citrus x microcarpa) peels extracts,
and combination of ginger (Zinger officinale) and (citrus x microcarpa) peels extracts
in terms of zone of inhibition.
Data Collection Procedure
Figure 3 shows the flowchart of the data collection procedure.
A.Collection of
Materials
B. Preparation of
Calamansi and Ginger
Peels
C. Ethanol Extraction
of the Calamansi and
Ginger Peels
F. Applying WoundInfecting Bacteria to
the Growth Medium
E. Obtaining the
bacteria
D. Preparation of
Agar Growth Solution
and Agar Growth
Medium
G. Data Collection
Research Ethics
H. Statistical
Treatment
In this study, the researchers took precautions to ensure that the results were not
biased. For obtaining the wound-infecting bacteria specimen, the researchers sought the
volunteer's agreement by providing a letter of agreement that outlined the purpose and
procedures of the study. During data collection, the researchers wore appropriate
personal protective equipment to ensure their safety.
Moreover, when utilizing the laboratory and apparatuses, the researchers
handled them with care, avoiding any damage. They also maintain the cleanliness of
the laboratory to prevent any contamination or interference with the results. By
adhering to these practices, the researchers aim to minimize bias and maintain the
integrity of the study.
Chapter IV
RESULTS AND DISCUSSION
This chapter presented the results and discussion of the data gathered
throughout the experiment. Results and discussions are ordered following the Statement
of the Problem of the study.
Problem 1. What is the antimicrobial activity of calamansi peels extracts in terms
of Zone of Inhibition?
Table 1. Zone of inhibition of calamansi peels extract.
Zone of Inhibition (mm)
Extracts
Calamansi
Extract
Replicates
Peels
Mean
A
B
C
D
E
2
0
0
2
0
0.8
Prescribed
Values
“+” <15mm
Table 1 showed the gathered data of zones of inhibition of calamansi peels
extracts against wound infecting bacteria. The mean for the treatment of calamansi
peels extract was 0.8mm which was prescribed as “+” <15mm, indicating a small effect.
This indicated that the calamansi peels extract applied to the agar medium
containing wound infecting bacteria had small effects. This finding supported the study
of Giron et al., (2017) which found that calamansi peels extracts exhibited antibacterial
activity against Staphylococcus aureus. It also aligned with the study of Trakranrungsie
and Chatchawanchonteera et al., (2016) which demonstrated that calamansi peels
extract showed significant antimicrobial activity against all tested foodborne pathogens.
Problem 2. What is the antimicrobial activity of ginger peels extracts in terms of
Zone of Inhibition?
Table 2. Zone of inhibition of ginger peels extract.
Zone of Inhibition (mm)
Extracts
Ginger Peels Extract
Replicates
Mean
A
B
C
D
E
0
0
0
0
0
0
Prescribed
Values
“-” no zone
Table 2 presented the gathered data of zones of inhibition of ginger peels
extracts against wound infecting bacteria. The mean for the treatment of calamansi
peels extract was 0mm which was prescribed as “-”, indicating no effect according to
Suflet et al., 2021.
This indicated that the ginger peels extract treatment applied to the agar medium
containing wound infecting bacteria had no effects. This contradicted the study of Islam
(2024) which demonstrated the potent antimicrobial activity of the ginger extract
against the all tested bacterial pathogens. However, a study of Abdullahi et al., (2019)
reviewed the literature on wound infections and antimicrobial resistance in small animal
practice and found that the emergence of antimicrobial-resistant bacterial strains has
become a major challenge in the treatment of wound infections in small animals. This
suggested that some bacterial pathogens have developed antimicrobial resistance to
different antimicrobial medications.
Problem 3. What is the antimicrobial activity of ginger and calamansi extracts in
terms of Zone of Inhibition?
Table 3. Zone of inhibition of calamansi and ginger peels extract.
Zone of Inhibition (mm)
Extracts
Calamansi
and
Ginger Peels Extract
Replicates
Mean
A
B
C
D
E
0
0
0
0
0
0
Prescribed
Values
“-” no zone
Table 3 presented the gathered data of zones of inhibition of calamansi and
ginger peels extracts against wound infecting bacteria. The mean for the treatment of
calamansi peels extract was 0mm which was prescribed as “-”, indicating no effect
according to Suflet et al., 2021.
This indicated that the calamansi and ginger peels extract treatment applied to
the agar medium containing wound infecting bacteria had no effects. This results
contradicted the study of Ongsakul et al. (2019) which investigated the antimicrobial
activity of Citrus microcarpa fruit extracts against some human pathogenic bacteria
and found that the extracts had the potential to be used as natural antimicrobial agents.
However, a research study by Abdullahi et al. (2019) reviewed the literature on wound
infections and antibiotic resistance in a small animal practice and discover that the
development of bacterial strains resistant to antibiotics has become a significant
obstacle in the management of wound infections in small animals. This demonstrates
how some bacterial infections have become resistant to various antimicrobial drugs.
Problem 4. What is the significant difference between the antimicrobial properties
of ginger (Zinger officinale) peels extract, calamansi (citrus x microcarpa) peels
extracts, and combination of ginger (Zinger officinale) and (citrus x microcarpa)
peels extracts in terms of zone of inhibition.
Table 4. One-Way ANOVA of different treatment groups.
Between
Groups
Sum of
Square
df
Mean
Squares
F
p-value
2.133
2
1.067
2.667
.110
Decision
on H0
Accept H0
Within
Groups
Total
4.800
6.933
12
.400
14
*significant at 0.05 level
Table 4 presented the results obtained from One-Way ANOVA, indicating a pvalue of 0.110. This was tested at alpha level 0.05 (F=2.667, p=0.110). Since the
computed p-value was greater than 0.05, the decision was to accept the null hypothesis,
suggesting no significant difference in the zone of inhibition among the three extracts.
Hence, their antimicrobial activity was closely similar to each other.
This indicated that calamansi peels extract, ginger peels extract, and calamansi
and ginger peels extract treatments applied to the agar medium containing bacteria
were closely similar in terms of their antimicrobial activity, as assessed by the zones of
inhibition.
Comparing the zone of inhibition among the three treatments, it was observed
that the calamansi peels extract treatment showed a noticeable zone of inhibition
compared to the other treatments performed. This result was consistent with the study
conducted by Bascon et al., (2020) which evaluated the antimicrobial activity of
calamansi against a range of pathogenic bacteria, including Salmonella typhimurium,
Staphylococcus aureus, and Escherichia coli. The researchers found that calamansi
extract exhibited strong antimicrobial activity against all the tested bacteria, suggesting
that calamansi could be a potential source of natural antimicrobial agents.
Furthermore, this result was supported by Mohammadifard et al. (2021) who
addressed the antibacterial activity of calamansi peel extracts against foodborne
pathogens. Their study found that the extracts exhibited significant antibacterial
activity, indicating their potential use as a natural alternative to synthetic antimicrobial
agents.
`
Chapter V
SUMMARY OF FINDINGS, CONCLUSIONS, AND RECOMMENDATIONS
This chapter presented the summary of findings, conclusions, and
recommendations.
Summary of Findings
The following were the findings of the study:
1. Antimicrobial Activity of Calamansi Peels extracts:
In terms of the antimicrobial activity of calamansi extracts, the zone of
inhibitions test results showed an observable zone of inhibitions for two
replicates (A, D) from 5 used replicates (A, B, C, D, E). This indicated that the
calamansi peels extracts had exhibited small antimicrobial activity against the
wound-infecting bacteria.
2. Antimicrobial Activity of Ginger Peels Extracts :
In terms of the antimicrobial activity of ginger extracts, the zone of
inhibitions test results showed no observable zones of inhibition for all
replicates (A, B, C, D, E). This indicates that ginger extracts did not exhibit
significant antimicrobial activity against the tested wound-infecting bacteria.
3. Antimicrobial Activity of Ginger Peels and Calamansi Peels Extracts:
In terms of the antimicrobial activity of ginger and calamansi extracts,
the zone of inhibitions test results showed no observable zones of inhibition for
all replicates (A, B, C, D, E). This suggested that the combination of ginger and
calamansi extracts did not enhance the antimicrobial activity against the tested
wound-infecting bacteria compared to individual extracts.
4. The significant difference between the antimicrobial properties of ginger
(Zingiber officinale) peels extract, calamansi (Citrus x microcarpa) peels
extract, and the combination of ginger and calamansi peels extracts in terms of
the zone of inhibition:
Based on the One-Way ANOVA analysis (Table 4), which aimed to
determine the significant difference between the treatment groups, the results
yielded a non-significant p-value of 0.110 at a significance level of 0.05
(F=2.667, p=0.110). Consequently, the null hypothesis was accepted, indicating
that there was no significant difference observed in the antimicrobial properties
of ginger peels extract, calamansi peels extract, and the combination of ginger
and calamansi peels extracts.
Conclusions
The researchers hereby conclude the following based on the findings of the
study:
1. The antimicrobial activity of calamansi peels extracts in terms of zone of
inhibition showed a zone of inhibition less than 15 mm, indicating a moderate
antimicrobial effect. Calamansi peels extract inhibited some level of inhibition
against the tested wound-infecting bacteria.
2. Based on the antimicrobial activity of ginger peels extracts in terms of zone of
inhibition, no zone of inhibition was observed, indicating that ginger extract did
not exhibit significant antimicrobial activity against the wound-infecting
bacteria tested.
3. When combining ginger peels and calamansi peels extracts, no zone of
inhibition was observed, suggesting that the combination did not enhance the
antimicrobial activity against the wound-infecting bacteria compared to
individual extracts.
4. To conclude, the results indicated that there was no significant difference in the
antimicrobial properties of ginger peels extract, calamansi peels extract, and the
combination of ginger and calamansi peels extracts in terms of the zone of
inhibition.
Nonetheless,
calamansi
peels
extract
showed
promising
antimicrobial activity, highlighting its potential as an effective natural
antimicrobial agent for combatting wound-infecting bacteria.
Recommendations
The following are the recommendations of the researchers:
1. Further study is required to fully explore the potential of these extracts as
antimicrobial agents in wound care, additional study is necessary to examine
the antibacterial activity of ginger and calamansi peel extracts against bacteria
that cause wound infection.
2. Conduct additional experiments with a wider variety of wound-infecting
bacterial strains to assess the range of antibacterial activity displayed by ginger
and calamansi peels.
3. Additional research should be done to locate and define the specific chemicals
found in these extracts that are responsible for their antibacterial activity in
order to clarify the underlying mechanism of action.
4. Choose pertinent wound-infecting bacterial strains for testing: It's critical to
select pertinent wound-infecting bacterial strains. Staphylococcus aureus,
Pseudomonas aeruginosa, Escherichia coli, and Enterococcus faecalis are
typical wound pathogens.
5. Invest in high-end laboratory apparatus to have more accurate findings in your
experiment.
6. Choosing a thinner filter paper disk to be used in disc diffusion is highly
recommended, for the reason that thicker filter papers absorb more and lesser
diffusion of extract due to its high absorbency.
7. Explore other alternative herbal medicines for antimicrobial activity and also
explore other extraction methods.
REFERENCES
Ayuob, N. N., El-Hawary, S. S., Ammar, S. M., & El-Sheekh, M. M. (2013).
Bioactivity and chemical composition of essential oils of four Egyptian
plants against antibiotic-resistant bacterial strains. Industrial Crops and
Products, 49, 83-87. https://doi.org/10.1016/j.indcrop.2013.04.040
Akinmoladun AC, Olaleye MT, Aiyegoro OA, et al. Chemical composition and
antibacterial activity of the essential oil of Syzygium aromaticum
(clove). Afr J Microbiol Res. 2018;12(6):130-135.
Bhatt, H. B., & Smith, R. J. (2012). Folic Acid Supplementation for the
Prevention of Neural Tube Defects: US Preventive Services Task Force
Recommendation
Statement.
JAMA,
307(1),
42–43.
https://doi.org/10.1001/jama.2011.1913
Chakraborty, D., & Ghosh, U. (2020). Antimicrobial activity of ginger
(Zingiber officinale) extract against foodborne pathogens and its
application in food systems: A review. Food Reviews International, 123. https://doi.org/10.1080/87559129.2020.1773464
Chakraborty, A., & Chatterjee, M. (2021). Comparative effectiveness of
Vitamin D3 and dietary Vitamin E on peroxidation of lipids and
enzymes of the hepatic antioxidant system in Sprague–Dawley rats.
International Journal for Vitamin and Nutrition Research, 67(1), 2-11.
https://doi.org/10.1024/0300-9831.67.1.2
Chantarasataporn, P., Kaewmanee, T., Lertsiri, S., & Lertsiri, S. (2020).
Evaluation of antimicrobial and antioxidant activities of calamansi
(Citrus microcarpa) peel extracts. Agriculture and Natural Resources,
54(5), 465-471. https://doi.org/10.34044/j.anres.2020.54.5.06
Ghasemi, Y., & Heydari, R. (2020). Ginger (Zingiber officinale) and its active
compounds as potential candidate for the prevention and treatment of
infection caused by Gram-positive bacteria: A review. Journal of
Essential
Oil
Research,
32(3),
189-208.
https://doi.org/10.1080/10412905.2020.1740642
Giron, L. M. P., Cabbuag, L. B., & Saplala, C. A. (2017). In vitro antibacterial
activity of Citrus microcarpa peel extract against Staphylococcus
aureus. Journal of Applied Biology & Biotechnology, 5(04), 62-67.
https://doi.org/10.7324/JABB.2017.50411
Jayaweera, S., Bhattarai, S., Tang, S. Y., & Toh, G. W. (2019). Development of
a low-cost topical antiseptic cream using natural extracts for wound
care.
Journal
of
Wound
Care,
28(8),
528-533.
https://doi.org/10.12968/jowc.2019.28.8.528
Jneid, J., Cassir, N., Schuldiner, S., Esseily, F., & Kempf, M. (2019). Antibiotic
resistance in bacterial pathogens and its potential implications in the
treatment of infected wounds. Frontiers in Microbiology, 10
Jones, L., Othman, M., Dowswell, T., Alfirevic, Z., Gates, S., Newburn, M.,
Jordan, S., Lavender, T., & Neilson, J. P. (2022). Pain management for
women in labour: an overview of systematic reviews. Cochrane
Database
of
Systematic
Reviews,
3,
CD009234.
https://www.cochrane.org/CD011712
Kaur, I. P., & Kakkar, S. (2015). Nanotherapy for posterior eye diseases. Journal
of
Controlled
Release,
206,
76-95.
https://doi.org/10.1016/j.jconrel.2015.03.025
Koffi-Nevry, R., Kouassi, K. C., & Kouamé, L. P. (2019). In vitro evaluation
of the antimicrobial activity of ginger (Zingiber officinale) and garlic
(Allium sativum) extracts against bacterial strains isolated from wound
infections. Journal of medicinal plants research, 13(2), 28-34.
Menniti-Ippolito, F., Da Cas, R., Traversa, G., Santuccio, C., Felicetti, P.,
Tartaglia, L., & Carle, F. (2022). Spontaneous reporting of adverse
reactions to herbal medicinal products in Italy: An analysis of the period
2002-2021.
PLoS
ONE,
16(11),
e0260137.
https://doi.org/10.1371/journal.pone.0260137
Mohammadifard, M., Akhavan Sepahi, A., Rahnama, M., & Tarighi, S. (2021).
Antibacterial activity of Calamansi (Citrus microcarpa) peel extracts
against foodborne pathogens. Journal of Food Safety, 41(2), e12818.
https://doi.org/10.1111/jfs.12818
National Academies of Sciences, Engineering, and Medicine. (2019). The
safety and quality of abortion care in the United States. National
Academies Press. https://www.ncbi.nlm.nih.gov/books/NBK507853/
Ongsakul, M., Chanthai, S., & Prangthip, P. (2019). Antimicrobial activity of
Citrus microcarpa fruit extracts against some human pathogenic
bacteria. Agriculture and Natural Resources, 53(4), 315-319.
https://doi.org/10.34044/j.anres.2019.53.4.02
Osho, A., Oguntoye, S. O., & Oyekunle, J. A. (2020). Antibacterial and
phytochemical properties of ginger (Zingiber officinale) against selected
foodborne
pathogens.
Heliyon,
6(4),
e03838.
https://doi.org/10.1016/j.heliyon.2020.e03838
Philippine Statistics Authority. (2023). Highlights of the 2019/2020 Integrated
Survey of Labor and Employment (ISLE) Module on Occupational
Injuries.
Retrieved
May
17,
2023,
from
https://psa.gov.ph/content/highlights-20192020-integrated-surveylabor-and-employment-isle-module-occupational-injuries
Pires, D., Diamantino, T., Santos, L., Fernandes, R., & de Pinho, P. G. (2019).
Wound infection and antimicrobial resistance in small animal practice:
A
review.
Veterinary
Sciences,
6(3),
74.
https://doi.org/10.3390/vetsci6030074
Rakholiya, K. D., Kaneria, M. J., & Chanda, S. V. (2013). Medicinal plants
as alternative sources of therapeutics against multidrug-resistant
pathogenic microorganisms based on their antimicrobial
potential and synergistic properties. In Fighting Multidrug
Resistance with Herbal Extracts, Essential Oils and Their
Components (pp. 165–179). Elsevier.
Rajesh, E., & Sankari, L. S. (2019). Antimicrobial activity of ginger extract
against oral biofilm. In E. Rajesh & L. S. Sankari (Eds.), Ginger
Cultivation and Its Antimicrobial Activity (pp. 101-110). Springer.
https://books.google.com.ph/books?id=AGH9DwAAQBAJ&pg=PA10
1
Sardar, S., Chakraborty, A., & Chatterjee, M. (2021). Comparative
effectiveness of Vitamin D3 and dietary Vitamin E on peroxidation of
lipids and enzymes of the hepatic antioxidant system in Sprague–
Dawley rats. International Journal for Vitamin and Nutrition Research,
67(1), 2-11. https://doi.org/10.1024/0300-9831.67.1.2
Shrestha, S., Malla, S., Maharjan, E., Karki, S., & Poudyal, P. (2020). Antibiotic
resistance in wound infection bacteria in a tertiary care hospital of
Nepal.
BMC
Research
Notes,
13(1),
1-7.
https://doi.org/10.1186/s13104-020-05023-1
Silversides, J. A., Lappin, E., & Ferguson, A. J. (2006). C-reactive protein
measurement in patients with unstable angina improves the prediction
of
cardiovascular
risk.
Heart,
92(6),
789-791.
https://doi.org/10.1136/hrt.2005.077578
Solberg, K. E. (2016). The WTO Agreement on Trade-Related Aspects of
Intellectual Property Rights and Access to Medicines. Asian Journal of
WTO & International Health Law and Policy, 11(1), 193-212.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4842230/
Speroff, T., Nwosu, S., Boutwell, A., & Costanza, M. E. (2018). Organisational
culture: variation across hospitals and connection to patient safety
climate. Quality & Safety in Health Care, 19(6), 592-596.
https://doi.org/10.1136/qshc.2009.039511
Suflet, D. M., Popescu, I., Pelin, I. M., Ichim, D. L., Daraba, O. M., Constantin,
M., & Fundueanu, G. (2021). Dual Cross-Linked Chitosan/PVA
Hydrogels Containing Silver Nanoparticles with Antimicrobial
Properties.
Pharmaceutics,
13(9),
1461.
https://doi.org/10.3390/pharmaceutics13091461
Trakranrungsie, N., & Chatchawanchonteera, A. (2016). Antimicrobial activity
of Citrus hystrix DC. (Kaffir lime) peel extract against foodborne
pathogens and application in Thai fish cake. LWT - Food Science and
Technology, 65, 557-563. https://doi.org/10.1016/j.lwt.2015.08.027
Vaismoradi, M., Salsali, M., Ahmadi, F. (2022). Patient Safety: Nursing
Students’ Perspectives and the Role of Nursing Education to Provide
Safe Patient Care. International Journal of Nursing Practice, 18(5), 471479. https://doi.org/10.1111/j.1440-172X.2012.02061.x
Villaflores, O. B., Chen, Y. S., Chen, Y. K., et al. (2017). Antimicrobial Activity
of Calamansi (Citrus microcarpa) Fruit Extracts and Essential Oils
against Foodborne Pathogens. Journal of Food Protection, 80(3), 434440. https://doi.org/10.4315/0362-028X.JFP-16-369
Zahoor, M., Jan, M. S., Hanif, M. A., Bilal, M., & Rahim, F. (2020). Chemical
composition and antimicrobial activity of essential oil of Zingiber
officinale against bacterial pathogens. Pakistan Journal of
Pharmaceutical Sciences, 33(3)
APPENDIX A: Production of Extracts and Agar Growth Medium
A. Collection of Materials
B. Preparation of Calamansi and
Ginger Peels
C. Ethanolic Extractions of Peels
D. Open Surface Evaporation
E. Preparation of Agar Solution
F. Sterilization of Materials
G. Making of Agar Medium
H. Soaking of Sterilized Filter
Papers
I. Obtaining Bacteria
J. Applying Wound Infecting
Bacteria
K. Checking for Bacterial Growth
L. Adding of Filter Paper Discs
M. Measuring the Zone of Inhibition
APPENDIX : Gantt Chart
APPENDIX: Expenses
ITEM
QUANTITY
PRICE
Agar Powder
1 tube
Php 4,500.00
Inoculating Rods
30 pieces
Php 90.00
Sterile Swabs
10 pieces
Php 100.00
Ginger
2-kilo
Php 120.00
Calamansi
2-kilo
Php 200.00
Filter Paper
6 pieces
Php 120.00
Vials
6 pieces
Php 30.00
Gauze Pad
1 roll
Php 40.00
Cotton
1 tube
Php 60.00
Sterile Gloves
6 pairs
Php 120.00
TOTAL
-
Php 5,380.00
CURRICULUM VITAE
Nino Neil P. Tapiz
Purok Makuguihon, Fisherman’s Village,
Miputak, Dipolog City
ninotapiz@gmail.com
PERSONAL BACKGROUND
Date of Birth:
Place of Birth:
Citizenship:
Religion:
Height:
Weight:
Father’s Name:
Father’s Occupation:
Mother’s Name:
Mother’s Occupation:
Home Address:
Language Spoken:
Skill/s:
October 02, 2004
Dipolog City
Filipino
Roman Catholic
165 cm
50 kg
Albino R. Tapiz Jr.
Private Employee
Nilda P. Tapiz
Self Employee
Purok Makuguihon, Fisherman’s Village, Miputak,
Dipolog City
Cebuano
Graphic Designing, Journalism, Public Speaking
EDUCATION ATTAINMENT
Senior High School
Junior High School
Zamboanga del Norte High
School
National
High School (ZNNHS),
Turno, Dipolog City
Zamboanga del Norte High
School
National
High School (ZNNHS),
Turno, Dipolog City
2021 - 2023
2017 - 2019
Miputak National High
School
Miputak, Dipolog City
2019 - 2021
Dipolog Pilot School
Demonstration School,
Dipolog City
2011 - 2013
Miputak East Central
School
Miputak, Dipolog City
2013 -2017
Grade School
CURRICULUM VITAE
Dhina Elijah Lane D. Jaime
Malvar St.
Estaka, Dipolog City
jaimedhinaelijahlanedelacruz@gmail.com
PERSONAL BACKGROUND
Date of Birth:
Place of Birth:
Citizenship:
Religion:
Height:
Weight:
Father’s Name:
Father’s Occupation:
Mother’s Name:
Mother’s Occupation:
Home Address:
Language Spoken:
Skill/s:
February 19, 2005
Cebu City
Filipino
Roman Catholic
4’11 ft
49 kg
Joseph R. Jaime
N/A
Jurelyn D. Jaime
N/A
Malvar St. Estaka, Dipolog City
English; Filipino ( Bisaya)
Fluency in English and Filipino, Writing, Music
EDUCATION ATTAINMENT
Senior High School
Junior High School
Grade School
Zamboanga del Norte High
School National High
School (ZNNHS), Turno,
Dipolog City
Zamboanga del Norte
National High School
(ZNNHS), General Luna
Street, Dipolog City
East Central School,
Palawan Puerto Princesa
2021 - 2023
2017-2021
2011 - 2017
CURRICULUM VITAE
Aaron Jhan C Potoy
Andres Bonifacio Street
Bagting, Dapitan City
ajpotoy@gmail.com
PERSONAL BACKGROUND
Date of Birth:
Place of Birth:
Citizenship:
Religion:
Height:
Weight:
Father’s Name:
Father’s Occupation:
Mother’s Name:
Mother’s Occupation:
Home Address:
Language Spoken:
Skill/s:
February 21, 2005
Dipolog City
Filipino
Roman Catholic
172 cm
78 kg
Bonifacio R. Potoy Jr.
Engineer
Meriam C. Potoy
Office Worker
Andres Bonifacio Street, Bagting, Dapitan City
English and Filipino
Unidentified
Senior High School
2021 - 2023
Zamboanga del Norte
National High School
Turno Campus
High School Level
2017 - 2021
Jose Rizal Memorial State
University Main Campus
Grade School
2011 - 2017
Dapitan City Central
School
CURRICULUM VITAE
Mark Joshua E. Lorion
Taga, Municipality of Katipunan
marklorion143@gmail.com
PERSONAL BACKGROUND
Date of Birth:
Place of Birth:
Citizenship:
Religion:
Height:
Weight:
Father’s Name:
Father’s Occupation:
Mother’s Name:
Mother’s Occupation:
Home Address:
Language Spoken:
Skill/s:
December 10, 2004
Dipolog City
Filipino
Roman Catholic
160 cm
55 kg
Romeo B. Lorion
Deceased
Marjorie E. Lorion
OFW
Taga, Municipality of Katipunan
English and Filipino
Singing, Dancing, Drawing
Senior High School
2021 - 2023
Zamboanga del Norte
National High School
Turno Campus
High School Level
2017 - 2021
Zamboanga del Norte
National High School
Grade School
2011 - 2017
Dipolog Pilot
Demonstration School
CURRICULUM VITAE
Nathan Nyle Eddor C. Evardo
Central Brgy, Dipolog City
nathaneddor@gmail.com
PERSONAL BACKGROUND
Date of Birth:
Place of Birth:
Citizenship:
Religion:
Height:
Weight:
Father’s Name:
Father’s Occupation:
Mother’s Name:
Mother’s Occupation:
Home Address:
Language Spoken:
Skill/s:
May 2, 2004
Dipolog City
Filipino
Pentacostal
178cm
64kg
Eddor S. Evardo
Joana Paula C. Evardo
Bonifacio St. Central Brgy, Dipolog City
English and Filipino
Multi instrumental skill, sketching, cooking
Senior High School
2021 - 2023
Zamboanga del Norte
National High School
Turno Campus
High School Level
2017 - 2021
Zamboanga del Norte
National High School
Grade School
2011 - 2017
Dipolog Pilot
Demonstration School
CURRICULUM VITAE
Jv Franc Chris A. Abad
Dawo, Dapitan City
jvabad98@gmail.com
PERSONAL BACKGROUND
Date of Birth:
Place of Birth:
Citizenship:
Religion:
Height:
Weight:
Father’s Name:
Father’s Occupation:
Mother’s Name:
Mother’s Occupation:
Home Address:
Language Spoken:
Skill/s:
Dipolog City
Filipino
Roman Catholic
60kg
Raymund B. Abad
Chief Executive Officer
Jo Anne A. Abad
Admin Officer
Dawo, Dapitan City
English and Filipino
Multi instrumental skill, singing, ballet, pole dancing
Senior High School
2021 - 2023
Zamboanga del Norte
National High School
Turno Campus
High School Level
2017 - 2021
Jose Rizal Memorial State
University
Grade School
2011 - 2017
Dipolog Pilot
Demonstration School
CURRICULUM VITAE
Mary Joy B. Janolino
Gulayon, Dipolog City
janolinomaryjoy@gmail.com
PERSONAL BACKGROUND
Date of Birth:
June 26, 2005
Place of Birth:
Dipolog City
Citizenship:
Filipino
Religion:
Seventh day Adventist
Height:
148cm
Weight:
57kg
Father’s Name:
Marlon D. Janolino
Father’s Occupation:
Government Employee
Mother’s Name:
Jocelyn B. Janolino
Mother’s Occupation:
N/A
Home Address:
Gulayon, Dipolog City
Language Spoken:
English and Filipino
Skill/s:
Dancing, Drawing, Painting
Senior High School
2021 - 2023
Zamboanga del Norte
National High School
Turno Campus
High School Level
2017 - 2021
Zamboanga del Norte
National High School
Turno Campus
Grade School
2011 - 2017
Dipolog Pilot
Demonstration School
CURRICULUM VITAE
John Paul Acopiado
Address: Barangay Turno, Dipolog City
Email: jpacopiado2021@gmail.com
PERSONAL BACKGROUND
Place of Birth:
Dipolog City
Date of Birth:
July 29, 2003
Citizenship:
Filipino
Religion:
Roman Catholic
Height:
5’7 ft
Weight:
82 kg
Father’s Name:
Apolonio T. Acopiado
Occupation:
Watchman
Mother’s Name:
Nida A. Acopiado
Occupation:
Private Employee
Home Address:
Turno, Dipolog City
Language:
English; Filipino (Bisaya)
Skills:
Music
Senior High School
Zamboanga del Norte
National High School
2021-2023
Junior High School
Zamboanga del Norte
National High School
2017-2020
Grade School
Dipolog Pilot
Demonstration School
2011-2017