Phytochemical composition and antibacterial activity of selected medicinal plants in a local community in the Philippines

Abstract

A survey of medicinal plants in a local community in Guimbal, Iloilo, Philippines identified ten plant species namely, hagonoy (Chromolaena odorata), pitogo (Cycas rumphii), adgao (Premna odorata), labnog (Ficus leucantatoma), talus (Homalomena rubescens), sinaw-sinaw (Peperomia pellucida), palochina (Senna alata), badyang (Alocasia macrorrhizos); bagacay (Bambusa vulgaris) and karupi (Alpinia sp). From these plants, two least-studied medicinal plants, B. vulgaris and Alpinia sp. were determined of their phytochemical composition and antibacterial properties following standard procedures. Aqueous and ethanolic extracts were prepared from these plants and the antibacterial activity of the extracts against two Gram-negative bacteria, Aeromonas hydrophila and Vibrio harveyi, and a Gram-positive, spore-forming bacterium, Bacillus albus were determined using Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC). Phytochemical analyses of B. vulgaris and Alpinia sp., indicated the presence of alkaloids, carbohydrates, glycosides, phytosterols, flavonoids and phenols and tannins. All extracts of the two medicinal plants inhibited the growth of A. hydrophila at a concentration of 0.5 g ml-1. The MBC and MIC for B. vulgaris aqueous extract and Alpinia sp. ethanolic extract against V. harveyi showed inhibition at 0.5g ml-1 except for the MIC of B. vulgaris aqueous extract which was at 0.25 g ml-1. There was no inhibition of B. albus from all extracts of both medicinal plants. These two medicinal plants can be further explored as potential sources of ingredients for the development of novel antibacterial drugs particularly in inhibiting Gram-negative bacteria.

Full text

171
Abastillas et al.
Int. J. Biosci.
2023
RESEARCH PAPER
RESEARCH PAPER RESEARCH PAPER
RESEARCH PAPER
OPEN ACCESS
OPEN ACCESSOPEN ACCESS
OPEN ACCESS
Phytochemical composition and antibacterial activity of
selected medicinal plants in a local community in the
Philippines
Maeryll K
risna A. Abastillas
1
, Thessa Joy R. Bagolcol
1
, Pamela B. Buro
1
,
Mark Igor B. Gayoba
1
, Joylyn C. Segurada
1
, Edda Brenda S. Yerro
1
,
Mary Lou C. Arabaca
1
, James L. Torres
1
, Christopher Marlowe A. Caipang
*2
1
Department of Biology, College of Liberal Arts,
Sciences, and Education,
University of San Agustin, Iloilo City 5000, Philippines
2
Division of Biological Sciences, College of Arts and Sciences, University of the Philippines Visayas,
Miag-ao 5023, Iloilo, Philippines
Key words:
Anti
-
microbial, Drug
discovery, Botanical, Plant extracts, Phytochemicals
http://dx.doi.org/10.12692/ijb/22.2.171
-
18
2
Article published on
February
09
,
20
2
3
Abstract
A survey of medicinal plants in a local
community in Guimbal, Iloilo, Philippines identified ten plant species
namely, hagonoy (Chromolaena odorata), pitogo (Cycas rumphii), adgao (Premna odorata), labnog (Ficus
leucantatoma), talus (Homalomena rubescens), sinaw-sinaw (Peperomia pellucida), palochina (Senna alata),
badyang (Alocasia macrorrhizos); bagacay (Bambusa vulgaris) and karupi (Alpinia sp). From these plants, two
least-studied medicinal plants, B. vulgaris and Alpinia sp. were determined of their phytochemical composition
and antibacterial properties following standard procedures. Aqueous and ethanolic extracts were prepared from
these plants and the antibacterial activity of the extracts against two Gram-negative bacteria, Aeromonas
hydrophila and Vibrio harveyi, and a Gram-positive, spore-forming bacterium, Bacillus albus were determined
using Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC).
Phytochemical analyses of B. vulgaris and Alpinia sp., indicated the presence of alkaloids, carbohydrates,
glycosides, phytosterols, flavonoids and phenols and tannins. All extracts of the two medicinal plants inhibited
the growth of A. hydrophila at a concentration of 0.5 g ml
-1
. The MBC and MIC for B. vulgaris aqueous extract
and Alpinia sp. ethanolic extract against V. harveyi showed inhibition at 0.5g ml
-1
except for the MIC of B.
vulgaris aqueous extract which was at 0.25 g ml
-1
. There was no inhibition of B. albus from all extracts of both
medicinal plants. These two medicinal plants can be further explored as potential sources of ingredients for the
development of novel antibacterial drugs particularly in inhibiting Gram-negative bacteria.
*
Corresponding Author: Christopher Marlowe A. Caipang  cmacaipang@yahoo.com
International Journal of Biosciences | IJB |
ISSN: 2220-6655 (Print), 2222-5234 (Online)
http://www.innspub.net
Vol. 22, No. 2, p. 171-182, 2023

172
Abastillas et al.
Int. J. Biosci.
2023
Introduction
In many tropical countries, medicinal plants are often
used on a regular basis as an alternative or
supplement to prescribed medicines that are hard to
obtain. With over 13000 plant species present, the
Philippines is considered one of the most important
biodiversity hotspots in the world (Nuneza et al.,
2021). People in many rural areas in the country rely
on traditional medicinal plants as remedies for almost
all ailments due to their accessibility, availability and
cultural acceptability (Hussain et al., 2018). However,
the vast knowledge of the economic and medical use
of many plants is yet to be discovered for the
advancement and development of novelties in drugs.
Natural products or derivatives account for more than
a third of all Food and Drug Administration-approved
medications and 48.6% of all cancer drugs registered.
Medicinal plants are one of the notable sources of
natural products. Many of its organs (Sofowara et al.,
2013) contain several phytochemicals, such as
flavonoids, alkaloids, tannins, and terpenoids, which
possess anti-microbial and antioxidant properties
(Talib and Mahasneh, 2010). The discoveries on the
presence of different bioactive and anti-microbial
components of medicinal plants helped researchers
track the sources of new and more effective drugs.
According to Oladeji (2016), some drugs believed to
be obtained from medicinal plants are aspirin,
atropine, artemisinin, colchicine, digoxin, ephedrine,
morphine and physostigmine.
Moreover, published studies each year helped further
recognize a variety of plants as medicinal. This
includes botanical surveys that allow people to gain a
better understanding of the various plant species that
exist in the environment. Plant surveys are also
crucial because they serve as a foundation for new
discoveries about medicinal plant applications and
approved therapeutic actions. A listing prepared by
Carag and Buot (2017) from available published
literature recorded at least 1000 medicinal plant
species and the common ailments each species is
utilized for. This checklist is an indication that the
Philippines has a rich medicinal flora and
practitioners of folk medicine possess extensive
knowledge of the medicinal properties of these
various plants. However, the rapid land degradation,
accelerated forest destructions, loss of biological
diversity, access to modern medicine, exposure to
modern culture, mobility, and displacement of
communities may affect the traditional knowledge
and the variety of the medicinal plants that are
present in a local community (Cordero and Alejandro,
2021). It is, therefore, urgent to document these data
in those local communities before it is totally
forgotten. The local community of Camangahan in
Guimbal, Iloilo is home to a diverse range of plant
species. Some are currently regarded as therapeutic
plants, but others have yet to be discovered for their
antibacterial properties. However, there is a scarcity
of research studies on the various medicinal plants
that are present and used in the community.
Moreover, their medicinal and antibacterial
properties are not well-documented. Hence, these
prompted the researchers to conduct a survey of
medicinal plants within the locality and to determine
the phytochemical composition and antibacterial
activities of the least-studied medicinal plants during
the survey.
Materials and methods
Study area
The study was conducted in the municipality of
Guimbal. This is a coastal municipality that is located
in the south-western part of Iloilo province. It is
situated between the coordinates of 1250 57’
longitude and 100 39’ latitude. It is 29 kilometers (18
mi) from Iloilo City and has a total land area of 4,461
hectares (11,020 acres). The municipality consists of
33 barangays (local communities), 22 of which are
located outside the town center and 11 are located
within the town center. Camangahan, a community
that is located outside of the town center, is abundant
in medicinal plants, but some of which are unknown.
Data gathering
Due to COVID restrictions and to avoid physical
contact, the gathering of information about the
medicinal plants that are being used by the local

173
Abastillas et al.
Int. J. Biosci.
2023
community was done through phone interviews. A
total of 30 local informants were interviewed by
phone and were only asked to mention the medicinal
plants they used and their traditional applications.
The names of the participants were kept anonymous
and personal information was not acquired. The
information gathered was treated with safety and
confidentiality.
Collection, enumeration, and description of
medicinal plants
Photographic data of uncommon medicinal plants
present was taken for proper photo documentation
and identification. The samples of these medicinal
plants were enumerated and identified using
published studies, personal interviews and
verification of botanists. Published literature were
also used to describe the botanical description,
phytochemical properties and medicinal uses of
plants.
Preparation of aqueous and ethanolic extracts
Preparation of both aqueous and ethanolic extracts
was done following the methods of Gonelimali et al.
(2018) with modifications. Prior to antibacterial
testing, the part of the plant tested was air-dried for
48 hours. The seeds of karupi (Alpinia sp.) were
utilized by the researchers, while the stems of bagacay
(Bambusa vulgaris) were used. The identities of these
plants were confirmed by the National Museum of
Natural History in Manila, Philippines, based on
close-up photos of the plant that showed its life
habits, the leaves, flowers and fruits that were sent
electronically. After air drying, the plant parts were
ground using a blender. In a clear container, 100 g of
powder from each tested plant material was soaked in
500 ml of distilled water and chilled for 24 hours.
Similarly, 100 g powder of each tested plant material
was soaked separately in another container with 500
ml of 80% ethanol and chilled for 24 hours.
Liquid extracts obtained were separated from the
solid residue by filtration using a cheesecloth. The
filtrates were placed into a beaker and dried using a
water bath. The residues were weighed, dissolved in
1% normal saline solution (NSS) to a final
concentration of 2 g ml
-1
, which served as the stock
solution, and kept at -20
o
C until use for the analyses.
Phytochemical analysis
In order to determine the phytochemical properties of
Bambusa vulgaris and Alpinia sp., the researchers
utilized the methods of Khalid et al. (2018) and
Shaikh and Patil (2020) with slight modifications.
The homogenate from the liquid extracts was used for
the analysis. The tests for the presence of alkaloids,
carbohydrates, glycosides, phytosterols, flavonoids,
phenols and tannins were carried out.
Preparation of inoculum
The antibacterial properties of medicinal plant
extracts were tested against two Gram-negative
bacteria: Aeromonas hydrophila and Vibrio harveyi,
and Bacillus albus, a Gram-positive, spore-forming
bacterium. The aforementioned broth cultures of
these bacteria were obtained from the Biology
Laboratory of the University of San Agustin. All
bacteria were cultured using Nutrient Broth (NB)
with the exception of V. harveyi, which was cultured
in NA with 1% sodium chloride.
After a 24-h incubation, ten-fold serial dilution of the
bacterial solutions was prepared and plated onto
Nutrient Agar medium and the bacterial count was
determined and expressed as colony-forming units
ml
-1
(CFU ml
-1
)). The bacterial solutions were diluted
to obtain a concentration of 1 × 10
4
CFU ml
-1
and were
utilized for the subsequent antibacterial assays.
Antibacterial assays
The minimum inhibitory concentration (MIC) was
determined using the broth microdilution method.
Each extract contained four different concentrations:
0.125g ml
-1
0.25g ml
-1
; 0.5g ml
-1
and 1g ml
-1
. Fifty
microliters (50 μl) of each extract with the specific
concentration was pipetted in each well of the 96 well
plates with three replicates. Each replicate was added
with an equal volume of the bacteria (A. hydrophila,
V. harveyi, B. albus) at a concentration of 1 × 10
4
CFU ml
-1
. The microplate was placed in an incubator

174
Abastillas et al.
Int. J. Biosci.
2023
at 28-30
o
C °C for 18–24 h. The control contained only
nutrient broth added with an equal volume of each
bacterium. MIC was determined as the lowest
concentration of either the aqueous and ethanolic
extract, where no visible growth of the bacteria was
observed in each well. The visual turbidity of the wells
was noted after incubation to confirm the MIC value.
The minimum bactericidal concentration (MBC) was
determined by the lowest concentration that inhibited
99.9% of bacterial growth. This was done by streaking
individual wells containing the mixtures of the plant
extracts and the bacteria onto the Nutrient Agar (NA)
plates for B. albus and A. hydrophila; and
Thiosulfate-Citrate-Bile Salts-Sucrose (TCBS) agar
plates for V. harveyi. The plates were incubated for
24 h at 28-30
o
C and observed for bacterial growth.
The lowest concentration that did not result in
bacterial growth was the MBC value for that
particular plant extract.
Results
Medicinal plants in Camangahan, Guimbal Iloilo
A total of ten (10) medicinal plants were identified
from the locality (Table 1). These included: Hagonoy
(Chromolaena odorata); Pitogo (Cycas rumphii);
Adgao; (Premna odorata); Labnog (Ficus
leucantatoma Merr.); Talus (Homalomena
rubescens (Roxb.) Kunth); Sinaw-sinaw (Peperomia
pellucida); Palochina (Senna Alata); Badyang
(Alocasia macrorrhizos); Bagacay (Bambusa
vulgaris) and Karupi (Alpinia sp).
Table 1. Enumeration and description of the medicinal plants in Camangahan, Guimbal, Iloilo based on
botanical description, phytochemical components and medicinal uses.
Medicinal Plant Botanical Description Phytochemical Components Medicinal Uses
Hagonoy (Chromolaena odorata)
Habit: A perennial shrub that grows up to 3–7 m zones
(Vijayaraghavan et al. 2017)
Leaves: Arrowhead-shaped (6–12 cm in length and 3–
7 cm in width) with three veins in pitchfork-shaped
appearance
Flowers: Have 5–25 tubular florets per head, each 10
mm long that are either white, purple, pink, or blue.
Seeds: brown-gray to black in color and is 4–5 mm
long (Sirinthipaporn and Jiraungkoorskul, 2017).
Leaves of this plant have been
found to be a rich source of
flavonoids saponin
triterpenoids, tannins, and
organic acids.
(Vijayaraghavan et al., 2017)
Wound healing, antibacterial,
antispasmodic, antiprotozoal,
antitrypanosomal, antifungal,
antihypertensive, anti-inflammatory,
astringent, diuretic, hepatotropic
immunomodulatory and anticancer
effects (Vijayaraghavan et al., 2017).
Pitogo
(Cycas rumphii)
Habit: A small tree or shrub that grows up to 10 m in
height with the trunk diameter reaches up to 400 mm.
Bark is gray with diamond and rectangular shape.
(Khan et al. 2011).
Leaves: 1.5-2.5 m long, ends with a paired glossy
pinnae or a spine 1-3 mm in length
Fruit: Sarcotesta has 3-4 mm thickness
Seeds: Green-orange color, 4.5-5 cm long, 3-3.5 cm in
diameter, flattened-ovoid shape (Hill, 1994).
Cycasin, β-glycosidase;
amentoflavone; podocarpus
flavone A, 2,3-dihydro
amentoflavone; 2,3-dihydro
hinoki flavone; isoginkgetin
and bilobetin (Khan et al.,
2011)
Effective for malignant ulcers, sore
throats, wounds healing, piles, boils,
itchy skin lesions, nephritic pains,
edematous swellings, dizziness,
headaches and tuberculosis (Khan et al.,
2011).
Adgao
(Premna odorata)
Habit: An evergreen small tree or shrub nearly 10 m
tall with diameter breast height ranging between 15–
30 cm.
Leaves: Leaves are opposite, ovate, hairy and green in
color, of 7–20 cm in length and 4–13.5 cm in width
Flowers:
flowers are pale green, yellowish or white that
are in inflorescences of 4–15 cm long
Fruits: globose drupe-like fruit with fleshy mericarps
(Youseff et al., 2021)
Alkaloids, anthraquinone,
saponins and steroids
(Mollejon and Mollejon,
2019).
Stomachache, headache, phlegm, cough,
and tuberculosis (Lirio et al., 2014).
Labnog
(Ficus septica)
Habit: A dioecious tree that grows 25 m tall and 20 cm
in diameter with a smooth, gray bark
Leaves: The leaves are smooth and shining, oblong-
ovate to elliptic ovate, margin is entire, 10-20 cm in
length with sharp point and pointed base.
Tannins, alkaloids, 2-deoxy
sugars, quaternary base, and
benzopyrone nucleus
(Vital et al., 2010)
Used as diuretic, analgesic, laxative, anti-
microbial, and antifungal. It is also
effective in treating fever, colds,
diarrhea, and cough (Jangad and
Licardo, 2018; Haryanti et al., 2021)

175
Abastillas et al.
Int. J. Biosci.
2023
Flowers: uniovulate female flowers and male flowers in
single or in pairs and are axillary (Conchou 2014).
Fruits: 1 mm long with tubercles (Mustaqim 2020)
Seeds: Orbicular cotyledons, present in female plants
but absent in males. (Conchou et al., 2014).
Talus
(Homalomena rubescens (Roxb.)
Kunth)
Habit: A leafy herbaceous plant that grows 10 cm tall
Leaves: Green-gray color, triangular, 8-10 cm long, 3-5
cm wide, pinnately netted vein
Flowers: Small, gray-brown color, 2-5 inflorescences
Fruits: absent
Seeds: Ellipsoid or elongate, endosperm copious,
embryo axile, significantly costate (Van et al., 2021).
Monoterpene hydrocarbons,
sesquiterpenes (Van et al.,
2021).
Antibacterial, and antioxidant
(Van et al., 2021).
Sinaw-sinaw
(Peperomia pellucida)
Habit: A herb that grows branched and upwar d up to 4
cm. Stems are succulent, round, and 5 mm thick. Spikes
are present with green pigment, slender, and 1-6 cm long.
Leaves: Heart-
shaped with alternate pattern, smooth, and
transparent.
Flowers: arranged in spike inflorescence about 2-6 cm
long, enclosed by round bracts
Fruits: Dry, indehiscent, round, 0.5 mm wide
Seeds: Dot-like appearance and attached to some fruiting
spikes (Ooi et al., 2012).
Secoligans, phytosterols,
tetrahydrofuran lignans,
steroids, tannins, xanthone
glycoside carbohydrates,
flavonoids, apiols, and
triterpenoids (Ooi et al.,
2012).
Used as treatment for gout and arthritis,
urinary tract inflammations,
constipation, kidney diseases, boils,
conjunctivitis, hypertension, tumors,
abscesses, breast cancer, convulsions,
and lowers cholesterol
(Ooi et al., 2012).
Palochina
(Senna Alata)
Habit: A shrub that produces flowers, grows 1-4 m tall,
proliferating in humid areas.
Leaves: The leaves are oblong that consists 5 to 14 leaflet
sets, intertwined bracts, and strong petioles that are 2-3
mm long.
Flowers: Condensed, bright yellow in color, with 7
stamens, and a ovary
Fruit: Thick, 10 x 15 in size, crystal-like pod, brown
Seeds: Brown and diamond-shaped (Oladeji, 2020).
Flavonoids, alkaloids,
anthraquinone,
cannabinoid, phenolics,
tannins, terpene, and
saponins
(Oladeji, 2020).
Used to treat diabetes, typhoid,
ringworms, scabies, malaria, asthma,
eczema, herpes, blotch, hepatitis,
gastroenteritis, syphilis, and gut
parasitosis (Oladeji, 2020).
Badyang
(Alocasia macrorrhizos)
Habit: An erect perennial, rhizomatous, monoecious
plant with short trunk that grows up to 5 m tall
Leaves: Large, ovate and can grow 1.8 m long and 1.2 m
wide, apex is pointed, cordate base, margins are slightly
undulate
Flowers: Green to white spathe encloses a greenish
spadix, inflorescences are large and form in clusters
Fruits: Green to scarlet berries, fleshy, 8 cm long,
ellipsoid or ovoid
Seeds: reddish, grows on the spadix (Lim, 2015).
Alkaloids, oxalic acid,
alocasins A-E, oleic acid,
linoleic acid, B-lectins,
ascorbic acid, and
cyanogenic glycosides
(Lim, 2015).
Serve as treatment for toothache,
abdominal pains, influenza, diarrhea,
malaria, tuberculosis, typhoid fever,
headaches, inflammations, rheumatism,
diabetes, etc.
(Lim, 2015).
Bagacay (Bambusa vulgaris)
Habit: An erect plant that can grow 20 m tall and 10 cm
in diameter. Stems are yellowish or yellowish-green
Leaves: Leaves have spikelets that are oblong and
clustered near the branches of inflorescence, lanceolate,
35 cm long and 4 cm wide
Flowers: Light-brown, rare
Fruits: Rare or absent due to irregular meiosis
Seeds: Produces in large amount in 1-3 years and
eventually dies
(Zheng et al., 2020).
Carbohydrates, glycosides,
saponins, alkaloids,
flavonoids, phenolics and
tannins, phytosterols, and
triterpenoids (Fitri et al.,
2020).
Used for kidney diseases, fever, diarrhea,
inflammations, measles, gonorrhea,
tuberculosis, wounds, and ulcers
(Owolabi and Lajide, 2015).

176
Abastillas et al.
Int. J. Biosci.
2023
Karupi (alpinia sp.)
Habit: A perennial herb with stems that are 1-2 m tall and
2.5 cm in diameter.
Leaves: Oblong-lanceolate, aromatic, root tubers
Flowers: The flower has a hollow toothed calyx that often
split on one side
Fruits: Round, red berries
Seeds: obtusely angular
(Chouni and Paul, 2018)
Tannins, glycosides,
phenols, carbohydrates,
monoterpenes, sterols,
sesquiterpenes, and
flavonoids such as
quercetin, kaempferol,
alpinin, and galangin
(Chouni and Paul, 2018)
Used for fungal skin disorders,
antirheumatic, dyspepsia, ulcers,
bronchitis, diabetes, obesity,
inflammations, diuretic, chest pains, sore
throat, and kidney diseases
(Chouni and Paul, 2018).
Phytochemical components of B. Vulgaris and
Alpinia sp.
Table 2 shows the phytochemical compounds present
in both karupi (Alpinia sp.) and bagacay (Bambusa
vulgaris). Both plants were positive for alkaloids,
carbohydrates, glycosides, phytosterols, flavonoids
and phenols and tannins.
Minimum Inhibitory Concentration (MIC) and
Minimum Bactericidal Concentrations (MBC) of
various plant extracts
MIC of various extracts against A. hydrophila
Table 3 shows the result of MIC of aqueous and
ethanolic extracts of B. vulgaris and Alpinia sp.
against A. hydrophila. Turbidity was not observed in
all extracts at concentrations of 0.5g ml
-1
and 1g ml
-1
,
indicating that bacterial growth was inhibited. Thus,
the lowest concentration of extracts that can inhibit
the growth of bacteria is 0.5 g ml
-1
.
MBC of various extracts against A. hydrophila
Absence in the growth of A. hydrophila was observed
in all extracts at concentrations of 0.5g ml
-1
and 1g ml
-
1
. Thus, the lowest concentration of the extracts that
can inhibit the growth of the bacteria is 0.5g/ml
(Table 4).
Table 2. Phytochemical compounds present in B. vulgaris and A. graminea.
Aqueous Sample Alkaloids Carbohydrates Glycosides Phytosterols Flavonoids Phenols and Tannins
B. vulgaris + + + + + +
Alpinia sp. + + + + + +
Positive (+): phytochemical is present; Negative (–): phytochemical is absent.
MIC of various extracts against V. harveyi
The MIC of the aqueous and ethanolic extracts of B.
vulgaris as well as the aqueous extract of Alpinia sp.
against V. harveyi is 0.25g ml
-1
. On the other hand,
the MIC of the ethanolic extracts of Alpinia sp. is 0.5g
ml
-1
(Table 5).
MBC of various extracts against V. harveyi
The MBC of the ethanolic extract of B. vulgaris and
aqueous extract of Alpinia sp. against V. harveyi was
0.25g ml
-1
. However, it was at 0.5 g ml
-1
for the
aqueous extract of B. vulgaris and ethanolic extract of
Alpinia sp. (Table 6).
Table 3. Minimum Inhibitory Concentrations (MIC) of aqueous (aq.) and ethanolic (eth.) extracts against A.
hydrophila.
Extract
Concentration
1g ml
-1
0.5g ml
-1
0.25g ml
-1
0.125g ml
-1
A.
B. vulgaris
(aq.)
-
-
+
+
B.
B. vulga
ris
(eth.)
-
-
+
+
C.
Alpinia sp
. (aq.)
-
-
+
+
D.
Alpinia sp.
(eth.)
-
-
+
+
Positive (+): Turbidity indicating growth; Negative (–): No turbidity indicating absence of growth.
MIC of various extracts against B. albus
The turbidity was present in all concentrations of
ethanolic and aqueous extracts indicating positive
results for bacterial growth. There was no inhibition
in all concentrations for Bacillus albus (Table 7).
MBC of various extracts against B. albus
The presence of bacterial growth was observed in all

177
Abastillas et al.
Int. J. Biosci.
2023
extracts. Thus, at all concentrations, there was no
inhibition of bacterial growth (Table 8).
Discussion
A total of ten (10) medicinal plants including
Hagonoy (Chromolaena odorata); Pitogo (Cycas
rumphii); Adgao; (Premna odorata); Labnog (Ficus
septica.); Talus (Homalomena rubescens (Roxb.)
Kunth); Sinaw-sinaw (Peperomia pellucida);
Palochina (Sena alata); Badyang (Alocasia
macrorrhizos); Bagacay (Bambusa vulgaris) and
Karupi (Alpinia sp.) were identified from the local
community. The majority of these plants were
classified as shrubs, herbs, trees, or erect plants.
Table 4. Minimum Bactericidal Concentrations (MBC) of aqueous (aq.) and ethanolic (eth.) extracts against A.
hydrophila.
Extract
Concentration
1g ml
-1
0.5g ml
-1
0.25g m
l
-1
0.125g ml
-1
A.
B. vulgaris
(aq.)
-
-
+
+
B.
B. vulgaris
(eth.)
-
-
+
+
C.
Alpinia sp
. (aq.)
-
-
+
+
D.
Alpinia sp.
(eth.)
-
-
+
+
Positive (+): Indicating growth; Negative (–): Indicating absence of growth.
Table 5. Minimum Inhibitory Concentrations (MIC) of aqueous (aq.) and ethanolic (eth.) extracts against V.
harveyi.
Extract
Concentration
1g ml
-1
0.5g ml
-1
0.25g ml
-1
0.125g ml
-1
A.
B. vulgaris
(aq.)
-
-
-
+
B.
B. vulgaris
(eth.)
-
-
-
+
C.
Alpinia sp
. (aq.)
-
-
-
+
D.
Alpinia sp.
(
eth.)
-
-
+
+
Positive (+): Turbidity indicating growth; Negative (–): No turbidity indicating absence of growth.
The morphology of the leaves, flowers, fruits, and
seeds varies among the ten plants identified.
Phytochemicals such as flavonoids, triterpenoids,
tannins, alkaloids, anthraquinone, saponins,
phytosterols, and phenolics have been found in these
plants (Vital et al., 2010; Vijayaraghavan et al., 2017;
Chouni and Paul, 2018). In addition, C. odorata, F.
septica, and H. rubescens have also been identified to
have antibacterial, antifungal and anticancer effects
(Vijayaraghavan et al., 2017; Jangad and Licardo,
2018; Haryanti et al., 2021; Van et al., 2021).
Furthermore, C. odorata, F. septica, P. pellucida, S.
alata, A. macrorrhizos, and B. vulgaris have are
regarded as diuretic, analgesic, laxative, anti-
inflammatory agent and has been used for treating
many illnesses including headache, stomachache,
urinary tract infections, tuberculosis and diabetes
(Ooi et al., 2012; Vijayaraghavan et al., 2017, Jangad
and Licardo, 2018; Haryanti et al., 2021).
Table 6. Minimum Bactericidal Concentrations (MBC) of aqueous (aq.) and ethanolic (eth.) extracts against V.
harveyi.
Extract
Concentration
1g ml
-1
0.5g ml
-1
0.25g ml
-1
0.125g ml
-1
A.
B. vulg
aris
(aq.)
-
-
+
+
B.
B. vulgaris
(eth.)
-
-
-
+
C.
Alpinia sp
. (aq.)
-
-
-
+
D.
Alpinia sp.
(eth.)
-
-
+
+
Positive (+): Indicating growth; Negative (–): Indicating absence of growth.

178
Abastillas et al.
Int. J. Biosci.
2023
The phytochemical analysis of B. vulgaris and Alpinia
sp. revealed the presence of alkaloids, carbohydrates,
glycosides, phytosterols, flavonoids and phenols and
tannins. The presence of phytochemicals has also
been found in the same genus of Alpinia sp. which is
Alpinia officinarum. It was stressed in the study by
Balamurugan et al. (2019) that the alcoholic extracts
with the use of established screening methods,
discovered the presence of phenols, tannins,
alkaloids, flavonoids, steroids, and quinones. In the
study of Fitri et al. (2020), B. vulgaris has been
reported to contain carbohydrates, glycosides,
saponins, alkaloids, flavonoids, phenolics and
tannins, phytosterols, and triterpenoids.
Table 7. Minimum Inhibitory Concentrations (MIC) of aqueous (aq.) and ethanolic (eth.) extracts against B.
albus.
Extract
Concen
tration
1g ml
-1
0.5g ml
-1
0.25g ml
-1
0.125g ml
-1
A.
B. vulgaris
(aq.)
+
+
+
+
B.
B. vulgaris
(eth.)
+
+
+
+
C.
Alpinia sp
. (aq.)
+
+
+
+
D.
Alpinia sp.
(eth.)
+
+
+
+
Positive (+): Turbidity indicating growth; Negative (–): No turbidity indicating absence of growth.
Both the minimum inhibitory concentration (MIC)
and minimum bactericidal concentration (MBC)
results of B. vulgaris and Alpinia sp. against A.
hydrophila, a Gram-negative bacterium, indicated
minimum inhibition of all extracts at 0.5g ml
-1
. In V.
harveyi, all extracts except ethanolic Alpinia sp.
exhibited a MIC at 0.25 g ml
-1
. The MBC for the
ethanolic extract of B. vulgaris and aqueous extract of
Alpinia sp. was 0.25g ml
-1
, whereas the aqueous
extract of B. vulgaris and ethanolic extract of Alpinia
sp. was at 0.5g ml
-1
. This clearly demonstrates that B.
vulgaris and Alpinia sp. are effective antibacterial
agents against these two gram-negative bacteria, A.
hydrophila and V. harveyi.
Previous studies have revealed that bamboo possesses
antioxidant, anticancer and anti-microbial activities.
Bioactive compounds found in B. vulgaris, a bamboo
species, include alkaloids, flavonoids, saponins, and
tannins that contribute to its antibacterial activity
(Tanaka et al., 2014). In the previous findings of
Zhiang et al. (2019), ethanolic extracts from leaves of
B. vulgaris have been found to exert minimum
inhibitory effects against a variety of bacterial species,
namely S. aureus and B. subtilis, at a concentration of
20g ml
-1.
(Zhiang et al., 2019). Additionally, the
ethanolic extracts and hot aqueous extracts of B.
vulgaris showed MIC values ranging at
concentrations 31.25 - 125 g ml
-1
against several
gastro-intestinal microorganisms such as E.coli, K.
pneumoniae, P. mirabilis, and S. typhi in the study of
Ogu et al. (2011).
The phytochemical compounds present in Alpinia sp.,
such as tannins, alkaloids, flavonoids and saponins,
have been detected to possess antibacterial activity
(Rini et al. 2018). The same phytochemicals were
found to be present in the study which largely
contributes to the potent antibacterial activity of the
aqueous and ethanolic extracts of Alpinia sp. against
A. hydrophila and V. harveyi. In the study of
Oonmetta-aree et al. (2006), among the four plants
tested, the ethanolic extracts of Alpinia galanga
demonstrated the highest inhibitory activity against
S. aureus.
The findings of the study showed the minimum
inhibitory concentration (MIC) of the ethanolic
extract at 0.325 mg ml
-1
and the minimum
bactericidal concentration (MBC) at 1.3 mg ml
-1
using
the broth diluteion method). However, in the study of
Voravuthikunchai et al. (2005), aqueous extracts of A.
galanga did not show any inhibitory activities against
S. aureus.

179
Abastillas et al.
Int. J. Biosci.
2023
Table 8. Minimum Bactericidal Concentrations (MBC) of aqueous (aq.) and ethanolic (eth.) extracts against B.
albus.
Extract
Concentration
1g ml
-1
0.5g ml
-1
0.25g ml
-1
0.125g ml
-1
A.
B. vulgaris
(aq.)
+
+
+
+
B.
B. vulgaris
(eth.)
+
+
+
+
C.
Alpinia sp
. (aq.)
+
+
+
+
D.
Alpinia sp.
(eth.)
+
+
+
+
Positive (+): Indicating growth; Negative (–): Indicating absence of growth.ive (–): Indicating absence of growth.
The results of MIC and MBC for B. albus are not
similar to the two Gram-negative bacteria
mentioned previously. B. albus is a spore-forming,
Gram-positive, facultative anaerobe bacterium. The
presence of turbidity in all extracts indicates
bacterial growth. The reason for this is that spore-
forming bacterium is resistant to heat, chemicals,
and other agents, making them tough to kill. This
explains why extracts obtained from Alpinia sp. and
B. vulgaris are ineffective antibacterial agents
against B. albus. as these bioactive substances are
not able to penetrate the thick cell wall components
of the spores. In a related study, Bacillus subtilis
which is also a Gram-positive spore-forming
bacteria, was used as a test organism for
determining the antibacterial activity of B. vulgaris
(Zhiang et al., 2019). However, the results indicated
that B. vulgaris was found to be an effective
antibacterial agent at a concentration of 20g ml
-1
. It
is most likely that Gram-positive bacteria are
inhibited by extracts from medicinal plants at higher
doses, and these will be explored in future studies.
Conclusion
The results from this study showed that aqueous and
ethanolic extracts from B. vulgaris and Alpinia sp.
contained phytochemical components that possess
potent bactericidal activities against Gram-negative
bacteria, A. hydrophila and V. harveyi. These two
least-studied medicinal plants can be further explored
as potential sources of bioactive substances for the
development of novel antibacterial drugs.
Acknowledgment
The authors greatly acknowledge the support
provided by the Friar administrators of the University
of San Agustin and the technical assistance of the
laboratory staff at the Department of Biology, College
of Liberal Arts, Sciences, and Education of the same
university.
References
Balamurugan V, Velurajan S, Palanisamy A.
2019. Phytochemical analysis of Alpinia officinarum
and to test its anti -oxidant, anti-microbial, anti-
inflammatory and anti -arthritic activity.
International Journal of Advance Research and
Innovative Ideas in Education 5, 1125-1140.
Carag H, Buot Jr IE. 2017. A checklist of the orders
and families of medicinal plants in the Philippines.
Sylvatrop, the Technical Journal of Philippine
Ecosystems and Natural Resources 27(1&2), 49-9.
Chouni A, Paul S. 2018. A review on phytochemical
and pharmacological potential of Alpinia galanga.
Pharmacognosy Journal 10(1), 09-15.
http://dx.doi.org/10.5530/pj.2018.1.2.
Conchou L, Cabioch L, Rodriguez LJV,
Kjellberg F. 2014. Daily rhythm of mutualistic
pollinator activity and scent emission in Ficus
septica: ecological differentiation between co-
occurring pollinators and potential consequences for
chemical communication and facilitation of host
speciation. PloS ONE 9(8), e103581.
https://doi.org/10.1371/journal.pone.0103581.

180
Abastillas et al.
Int. J. Biosci.
2023
Cordero C, Alejandro GJ. 2021. Medicinal plants
used by the indigenous Ati tribe in Tobias Fornier,
Antique, Philippines. Biodiversitas Journal of
Biological Diversity 22(2).
https://doi.org/10.13057/biodiv/d220203.
Fitri A, Asra R, Rivai H. 2020. Overview of the
traditional, phytochemical and pharmacological uses
of gold bamboo (Bambusa vulgaris). World Journal
of Pharmacy and Pharmaceutical Sciences 9(8), 21.
Gonelimali FD, Lin J, Miao W, Xuan J,
Charles F, Chen M, Hatab SR. 2018.
Antimicrobial properties and mechanism of action of
some plant extracts against food pathogens and
spoilage microorganisms. Frontiers in Microbiology
9, 1639.
https://doi.org/10.3389/fmicb.2018.01639.
Haryanti S, Rahmawati N, Sholikhah I,
Widiyastuti Y. 2021. Ficus septica, an ecosystem
keystone species induced ROS-mediated cytotoxicity
in HepG2 hepatocarcinoma cells. Earth
Environmental Science 905(1), 012101.
http://dx.doi.org/10.1088/1755-1315/905/1/012101.
Hill KD. 1994. The Cycas rumphii complex
(Cycadaceae) in New Guinea and the western Pacific.
Australian Systematic Botany 7(6), 543-567.
https://doi.org/10.1071/SB9940543.
Hussain W, Badshah L, Ullah M. 2018.
Quantitative study of medicinal plants used by the
communities residing in Koh-e-Safaid Range,
Northern Pakistani-Afghan borders. Journal of
Ethnobiology and Ethnomedicine 14(1), 1-18.
https://doi.org/10.1186/s13002-018-0229-4.
Jangad AMA, Licardo ADMB. 2018. Acute and
chronic toxicity studies of lagnob (Ficus septica
Burm. F. 1768) fruit extract on albino rats (Rattus
norvegicus). Doctoral dissertation, Thesis. University
of San Carlos-Josef Baumgartner Learning Resource
Center. Talamaban, Cebu City, Philipines.
Khalid S, Shahzad A, Basharat N, Abubakar
M, Anwar P. 2018. Phytochemical screening and
analysis of selected medicinal plants in Gujrat.
Journal of Phytochemistry and Biochemistry 2(1), 1-
3.
Khan AV, Ahmed QU, Khan AA, Shukla I. 2011.
Antibacterial activity of Cycas rumphii Miq. leaves
extracts against some tropical human pathogenic
bacteria. Research Journal of Microbiology 6(10),
761.
Lim TK. 2015. Alocasia macrorrhizos. Edible
Medicinal and Non Medicinal Plants: Volume 9,
Modified Stems, Roots, Bulbs. Springer, London.
429-442 p.
Lirio SB, Macabeo AP, Paragas EM, Knorn M,
Kohls P, Franzblau SG, Wang Y, Aguinaldo
MA. 2014. Antitubercular constituents from Premna
odorata Blanco. Journal of Ethnopharmacology 154
(2), 471-474.
https://doi.org/10.1016/j.jep.2014.04.015
Mollejon B, Mollejon C. 2019. Anti-microbial
activities and phytochemical screening of the Premna
odorata Blanco alagaw leaf extract. International
Journal of Trend in Scientific Research and
Development 3, 343-345.
Nuneza O, Rodriquez B, Nsaid JG. 2021.
Ethnobotanical survey of medicinal plants used by the
Mamanwa tribe of Surigao del Norte and Agusan del
Norte, Mindanao, Philippines. Biodiversitas 22(6).
https://doi.org/10.13057/biodiv/d220634.
Ogu GI, Madagwu EC, Eboh OJ. 2011.
Antibacterial activities of three medicinal plants
against some gastro-intestinal microorganisms.
International Journal of Natural and Applied Sciences
7(3), 248-256.
Oladeji O. 2016. The characteristics and roles of
medicinal plants: some important medicinal plants in
Nigeria. Natural Products: An Indian Journal 12(3),
102.

181
Abastillas et al.
Int. J. Biosci.
2023
Oladeji O, Adelowo F, Oluyori A, Bankole D.
2020. Ethnobotanical Description and Biological
Activities of Senna alata. Evidence-Based
Complementary and Alternative Medicine 2020,
2580259.
https://doi.org/10.1155/2020/2580259.
Ooi DJ, Iqbal S, Ismail M. 2012. Proximate
composition, nutritional attributes and mineral
composition of Peperomia pellucida L.
(Ketumpangan Air) grown in Malaysia. Molecules
(Basel, Switzerland) 17(9), 11139–11145.
https://doi.org/10.3390/molecules170911139.
Oonmetta-aree J, Suzuki T, Gasaluck P,
Eumkeb G. 2006. Antimicrobial properties and
action of galangal (Alpinia galanga Linn.) on
Staphylococcus aureus. Lebensmittel-Wissenschaft
Technologies-Food Science and Technology 39(10),
1214-1220.
https://doi.org/10.1016/j.lwt.2005.06.015.
Owolabi M, Lajide L. 2015. Preliminary
phytochemical screening and anti-microbial activity
of crude extracts of Bambusa vulgaris Schrad. Ex J.C.
Wendl. (Poaceae) from south-western Nigeria.
American Journal of Essential Oils and Products 3
(1), 42-45.
Rini C, Rohmah J, Widyaningrum L. 2018. The
antibacterial activity test galanga (Alpinia galangal)
on the growth of bacteria Bacillus subtilis and
Escherichia coli. IOP Conference Series: Materials
Science and Engineering 420, 012142.
http://dx.doi.org/10.1088/1757-899X/420/1/012142.
Sirinthipaporn A, Jiraungkoorskul W. 2017.
Wound healing property review of Siam weed,
Chromolaena odorata. Pharmacognosy Reviews 11
(21), 35–38.
http://dx.doi.org/10.4103/phrev.phrev_53_16.
Shaikh J, Patil M. 2020. Qualitative tests for
preliminary phytochemical screening: an overview.
International Journal of Chemical Study 8(2), 603-
608.
Sofowora A, Ogunbodede E, Onayade A. 2013.
The role and place of medicinal plants in the
strategies for disease prevention. African Journal of
Traditional, Complementary, and Alternative
Medicines 10(5), 210–229.
http://dx.doi.org/10.4314/ajtcam.v10i5.2.
Talib WH, Mahasneh AM. 2010. Anti-microbial,
cytotoxicity and phytochemical screening of
Jordanian plants used in traditional medicine.
Molecules 15(3), 1811-1824.
https://doi.org/10.3390/molecules15031811.
Tanaka A, Zhu Q, Tan H, Horiba H, Ohnuki K,
Mori Y, Yamauchi R, Ishikawa H, Iwamoto A,
Kawahara H, Shimizu K. 2014. Biological
activities and phytochemical profiles of extracts from
different parts of bamboo (Phyllostachys
pubescens). Molecules 19(6), 8238–8260.
https://doi.org/10.3390/molecules19068238
Van HT, Nguyen QP, Tran GB, Huynh NTA.
2021. Chemical composition and antibacterial
activities of Homalomena vietnamensis bogner & vd
nguyen (Araceae). Journal of Microbiology,
Biotechnology and Food Sciences 2021, 201-204.
http://dx.doi.org/10.15414/jmbfs.2020.10.2.201-204.
Vijayaraghavan K, Rajkumar J, Bukhari SN,
Al‑
‑‑
‑Sayed B, Seyed MA. 2017. Chromolaena
odorata: a neglected weed with a wide spectrum of
pharmacological activities (Review). Molecular
Medicine Reports 15, 1007-1016.
https://doi.org/10.3892/mmr.2017.6133.
Voravuthikunchai SP, Phongpaichit S,
Subhadhirasakul S. 2005. Evaluation of
antibacterial activities of medicinal plants widely used
among AIDS patients in Thailand. Pharmaceutical
Biology 43(8), 701-706.
https://doi.org/10.1080/13880200500385194.

182
Abastillas et al.
Int. J. Biosci.
2023
Vital P, Velasco R, Demigillo J, Rivera W.
2010. Antimicrobial activity, cytotoxicity and
phytochemical screening of Ficus septica Burm and
Sterculia foetida L. leaf extracts. Journal of Medicinal
Plants Research 4(1), 58-63.
http://dx.doi.org/10.5897/JMPR09.400.
Youssef FS, Ovidi E, Musayeib NMA, Ashour
ML. 2021. Morphology, anatomy and secondary
metabolites investigations of Premna odorata Blanco
and evaluation of its anti-tuberculosis activity using
in vitro and in silico studies. Plants 10(9), 1953.
https://doi.org/10.3390/plants10091953.
Zheng X, Lin S, Fu H, Wan Y, Ding Y. 2020. The
bamboo flowering cycle sheds light on flowering
diversity. Frontiers in Plant Sciences 11, 381.
https://doi.org/10.3389/fpls.2020.00381.
Zhiang K, Sari R, Apridamayanti P. 2019.
Minimum inhibitory concentration etermination of
Bambusa vulgaris leaves extract against skin and
digestive diseases bacteria. Kartika: Jurnal Ilmiah
Farmasi 7(1), 1-5.
http://dx.doi.org/10.26874/kjif.v7i1.158.