International Journal of Biosciences | IJB | ISSN: 2220-6655 (Print), 2222-5234 (Online) http://www.innspub.net Vol. 22, No. 2, p. 171-182, 2023 RESEARCH PAPER OPEN ACCESS !" % $ + & ' # ! ' ( $ ! + " $ !) !' ! ! ( * ! , ' $- !!! "# # "# $ & ' !$( % "# Key words: Anti-microbial, Drug discovery, Botanical, Plant extracts, Phytochemicals ! " # 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 vulgaris aqueous extract which was at 0.25 g ml -1. -1 except for the MIC of B. 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.  171 Abastillas et al. Introduction practitioners of folk medicine possess extensive In many tropical countries, medicinal plants are often knowledge of the medicinal properties of these used on a regular basis as an alternative or various plants. However, the rapid land degradation, supplement to prescribed medicines that are hard to accelerated forest destructions, loss of biological obtain. With over 13000 plant species present, the diversity, access to modern medicine, exposure to Philippines is considered one of the most important modern culture, mobility, and displacement of biodiversity hotspots in the world (Nuneza et al., communities may affect the traditional knowledge 2021). People in many rural areas in the country rely and the variety of the medicinal plants that are on traditional medicinal plants as remedies for almost present in a local community (Cordero and Alejandro, all ailments due to their accessibility, availability and 2021). It is, therefore, urgent to document these data cultural acceptability (Hussain et al., 2018). However, in those local communities before it is totally the vast knowledge of the economic and medical use forgotten. The local community of Camangahan in of many plants is yet to be discovered for the Guimbal, Iloilo is home to a diverse range of plant advancement and development of novelties in drugs. species. Some are currently regarded as therapeutic plants, but others have yet to be discovered for their Natural products or derivatives account for more than antibacterial properties. However, there is a scarcity a third of all Food and Drug Administration-approved of research studies on the various medicinal plants medications and 48.6% of all cancer drugs registered. that are present and used in the community. Medicinal plants are one of the notable sources of Moreover, natural products. Many of its organs (Sofowara et al., properties are not well-documented. Hence, these 2013) contain several phytochemicals, such as prompted the researchers to conduct a survey of flavonoids, alkaloids, tannins, and terpenoids, which medicinal plants within the locality and to determine possess anti-microbial and antioxidant properties the phytochemical composition and antibacterial (Talib and Mahasneh, 2010). The discoveries on the activities of the least-studied medicinal plants during presence of different bioactive and anti-microbial the survey. their medicinal and antibacterial components of medicinal plants helped researchers track the sources of new and more effective drugs. Materials and methods According to Oladeji (2016), some drugs believed to Study area be obtained from medicinal plants are aspirin, The study was conducted in the municipality of atropine, artemisinin, colchicine, digoxin, ephedrine, Guimbal. This is a coastal municipality that is located morphine and physostigmine. in the south-western part of Iloilo province. It is situated between the coordinates of 1250 57’ Moreover, published studies each year helped further longitude and 100 39’ latitude. It is 29 kilometers (18 recognize a variety of plants as medicinal. This mi) from Iloilo City and has a total land area of 4,461 includes botanical surveys that allow people to gain a hectares (11,020 acres). The municipality consists of better understanding of the various plant species that 33 barangays (local communities), 22 of which are exist in the environment. Plant surveys are also located outside the town center and 11 are located crucial because they serve as a foundation for new within the town center. Camangahan, a community discoveries about medicinal plant applications and that is located outside of the town center, is abundant approved therapeutic actions. A listing prepared by in medicinal plants, but some of which are unknown. Carag and Buot (2017) from available published literature recorded at least 1000 medicinal plant Data gathering species and the common ailments each species is Due to COVID restrictions and to avoid physical utilized for. This checklist is an indication that the contact, the gathering of information about the Philippines medicinal plants that are being used by the local has a rich 172 Abastillas et al. medicinal flora and community was done through phone interviews. A 1% total of 30 local informants were interviewed by concentration of 2 g ml-1, which served as the stock normal saline solution (NSS) to a final phone and were only asked to mention the medicinal solution, and kept at -20oC until use for the analyses. plants they used and their traditional applications. The names of the participants were kept anonymous Phytochemical analysis and personal information was not acquired. The In order to determine the phytochemical properties of information gathered was treated with safety and Bambusa vulgaris and Alpinia sp., the researchers confidentiality. utilized the methods of Khalid et al. (2018) and Shaikh and Patil (2020) with slight modifications. Collection, enumeration, and description of The homogenate from the liquid extracts was used for medicinal plants the analysis. The tests for the presence of alkaloids, Photographic data of uncommon medicinal plants carbohydrates, glycosides, phytosterols, flavonoids, present was taken for proper photo documentation phenols and tannins were carried out. and identification. The samples of these medicinal plants were using interviews and The antibacterial properties of medicinal plant verification of botanists. Published literature were extracts were tested against two Gram-negative also used to describe the botanical description, bacteria: Aeromonas hydrophila and Vibrio harveyi, studies, and Preparation of inoculum identified published enumerated personal phytochemical properties and medicinal uses of and Bacillus albus, a Gram-positive, spore-forming plants. bacterium. The aforementioned broth cultures of these bacteria were obtained from the Biology Preparation of aqueous and ethanolic extracts Laboratory of the University of San Agustin. All Preparation of both aqueous and ethanolic extracts bacteria were cultured using Nutrient Broth (NB) was done following the methods of Gonelimali et al. with the exception of V. harveyi, which was cultured (2018) with modifications. Prior to antibacterial in NA with 1% sodium chloride. testing, the part of the plant tested was air-dried for 48 hours. The seeds of karupi (Alpinia sp.) were After a 24-h incubation, ten-fold serial dilution of the utilized by the researchers, while the stems of bagacay bacterial solutions was prepared and plated onto (Bambusa vulgaris) were used. The identities of these Nutrient Agar medium and the bacterial count was plants were confirmed by the National Museum of determined and expressed as colony-forming units Natural History in Manila, Philippines, based on ml-1 (CFU ml-1)). The bacterial solutions were diluted close-up photos of the plant that showed its life to obtain a concentration of 1 × 104 CFU ml-1 and were habits, the leaves, flowers and fruits that were sent utilized for the subsequent antibacterial assays. electronically. After air drying, the plant parts were ground using a blender. In a clear container, 100 g of Antibacterial assays powder from each tested plant material was soaked in The minimum inhibitory concentration (MIC) was 500 ml of distilled water and chilled for 24 hours. determined using the broth microdilution method. Similarly, 100 g powder of each tested plant material Each extract contained four different concentrations: was soaked separately in another container with 500 0.125g ml ml of 80% ethanol and chilled for 24 hours. microliters (50 μl) of each extract with the specific -1 0.25g ml -1; 0.5g ml -1 and 1g ml -1. Fifty concentration was pipetted in each well of the 96 well Liquid extracts obtained were separated from the plates with three replicates. Each replicate was added solid residue by filtration using a cheesecloth. The with an equal volume of the bacteria (A. hydrophila, filtrates were placed into a beaker and dried using a V. harveyi, B. albus) at a concentration of 1 × 104 water bath. The residues were weighed, dissolved in CFU ml-1. The microplate was placed in an incubator 173 Abastillas et al. at 28-30oC °C for 18–24 h. The control contained only 24 h at 28-30oC and observed for bacterial growth. nutrient broth added with an equal volume of each The lowest concentration that did not result in bacterium. MIC was determined as the lowest bacterial growth was the MBC value for that concentration of either the aqueous and ethanolic particular plant extract. extract, where no visible growth of the bacteria was observed in each well. The visual turbidity of the wells Results was noted after incubation to confirm the MIC value. Medicinal plants in Camangahan, Guimbal Iloilo A total of ten (10) medicinal plants were identified The minimum bactericidal concentration (MBC) was from the locality (Table 1). These included: Hagonoy determined by the lowest concentration that inhibited (Chromolaena odorata); Pitogo (Cycas rumphii); 99.9% of bacterial growth. This was done by streaking Adgao; individual wells containing the mixtures of the plant leucantatoma (Premna odorata); Merr.); Labnog Talus (Ficus (Homalomena extracts and the bacteria onto the Nutrient Agar (NA) rubescens (Roxb.) Kunth); Sinaw-sinaw (Peperomia plates pellucida); Palochina Thiosulfate-Citrate-Bile Salts-Sucrose (TCBS) agar (Alocasia macrorrhizos); plates for V. harveyi. The plates were incubated for vulgaris) and Karupi (Alpinia sp). for B. albus and A. hydrophila; and (Senna Alata); Bagacay Badyang (Bambusa Table 1. Enumeration and description of the medicinal plants in Camangahan, Guimbal, Iloilo based on botanical description, phytochemical components and medicinal uses. Medicinal Plant Hagonoy (Chromolaena odorata) Botanical Description Phytochemical Components Habit: A perennial shrub that grows up to 3–7 m zones Leaves of this plant have been Medicinal Uses Wound healing, antibacterial, (Vijayaraghavan et al. 2017) found to be a rich source of antispasmodic, antiprotozoal, Leaves: Arrowhead-shaped (6–12 cm in length and 3– flavonoids saponin antitrypanosomal, antifungal, 7 cm in width) with three veins in pitchfork-shaped triterpenoids, tannins, and antihypertensive, anti-inflammatory, appearance organic acids. astringent, diuretic, hepatotropic Flowers: Have 5–25 tubular florets per head, each 10 (Vijayaraghavan et al., 2017) immunomodulatory and anticancer mm long that are either white, purple, pink, or blue. effects (Vijayaraghavan et al., 2017). Seeds: brown-gray to black in color and is 4–5 mm long (Sirinthipaporn and Jiraungkoorskul, 2017). Pitogo Habit: A small tree or shrub that grows up to 10 m in Cycasin, β-glycosidase; Effective for malignant ulcers, sore (Cycas rumphii) height with the trunk diameter reaches up to 400 mm. amentoflavone; podocarpus throats, wounds healing, piles, boils, Bark is gray with diamond and rectangular shape. flavone A, 2,3-dihydro itchy skin lesions, nephritic pains, (Khan et al. 2011). amentoflavone; 2,3-dihydro edematous swellings, dizziness, Leaves: 1.5-2.5 m long, ends with a paired glossy hinoki flavone; isoginkgetin headaches and tuberculosis (Khan et al., pinnae or a spine 1-3 mm in length and bilobetin (Khan et al., 2011). Fruit: Sarcotesta has 3-4 mm thickness 2011) Seeds: Green-orange color, 4.5-5 cm long, 3-3.5 cm in diameter, flattened-ovoid shape (Hill, 1994). Adgao Habit: An evergreen small tree or shrub nearly 10 m Alkaloids, anthraquinone, Stomachache, headache, phlegm, cough, (Premna odorata) tall with diameter breast height ranging between 15– saponins and steroids and tuberculosis (Lirio et al., 2014). 30 cm. (Mollejon and Mollejon, Leaves: Leaves are opposite, ovate, hairy and green in 2019). 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) Labnog Habit: A dioecious tree that grows 25 m tall and 20 cm (Ficus septica) in diameter with a smooth, gray bark sugars, quaternary base, and Leaves: The leaves are smooth and shining, oblong- benzopyrone nucleus effective in treating fever, colds, ovate to elliptic ovate, margin is entire, 10-20 cm in (Vital et al., 2010) diarrhea, and cough (Jangad and length with sharp point and pointed base. 174 Abastillas et al. Tannins, alkaloids, 2-deoxy Used as diuretic, analgesic, laxative, antimicrobial, and antifungal. It is also Licardo, 2018; Haryanti et al., 2021) 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 Habit: A leafy herbaceous plant that grows 10 cm tall Monoterpene hydrocarbons, Antibacterial, and antioxidant (Homalomena rubescens (Roxb.) Leaves: Green-gray color, triangular, 8-10 cm long, 3-5 sesquiterpenes (Van et al., (Van et al., 2021). Kunth) cm wide, pinnately netted vein 2021). Flowers: Small, gray-brown color, 2-5 inflorescences Fruits: absent Seeds: Ellipsoid or elongate, endosperm copious, embryo axile, significantly costate (Van et al., 2021). Sinaw-sinaw Habit: A herb that grows branched and upward up to 4 Secoligans, phytosterols, (Peperomia pellucida) cm. Stems are succulent, round, and 5 mm thick. Spikes tetrahydrofuran lignans, are present with green pigment, slender, and 1-6 cm long. steroids, tannins, xanthone Used as treatment for gout and arthritis, urinary tract inflammations, constipation, kidney diseases, boils, Leaves: Heart-shaped with alternate pattern, smooth, and glycoside carbohydrates, conjunctivitis, hypertension, tumors, transparent. flavonoids, apiols, and abscesses, breast cancer, convulsions, Flowers: arranged in spike inflorescence about 2-6 cm triterpenoids (Ooi et al., and lowers cholesterol long, enclosed by round bracts 2012). (Ooi et al., 2012). Fruits: Dry, indehiscent, round, 0.5 mm wide Seeds: Dot-like appearance and attached to some fruiting spikes (Ooi et al., 2012). Palochina Habit: A shrub that produces flowers, grows 1-4 m tall, Flavonoids, alkaloids, Used to treat diabetes, typhoid, (Senna Alata) proliferating in humid areas. anthraquinone, ringworms, scabies, malaria, asthma, Leaves: The leaves are oblong that consists 5 to 14 leaflet cannabinoid, phenolics, eczema, herpes, blotch, hepatitis, sets, intertwined bracts, and strong petioles that are 2-3 tannins, terpene, and gastroenteritis, syphilis, and gut mm long. saponins parasitosis (Oladeji, 2020). Flowers: Condensed, bright yellow in color, with 7 (Oladeji, 2020). stamens, and a ovary Fruit: Thick, 10 x 15 in size, crystal-like pod, brown Seeds: Brown and diamond-shaped (Oladeji, 2020). Badyang Habit: An erect perennial, rhizomatous, monoecious Alkaloids, oxalic acid, Serve as treatment for toothache, (Alocasia macrorrhizos) plant with short trunk that grows up to 5 m tall alocasins A-E, oleic acid, abdominal pains, influenza, diarrhea, Leaves: Large, ovate and can grow 1.8 m long and 1.2 m linoleic acid, B-lectins, malaria, tuberculosis, typhoid fever, wide, apex is pointed, cordate base, margins are slightly ascorbic acid, and headaches, inflammations, rheumatism, undulate cyanogenic glycosides diabetes, etc. Flowers: Green to white spathe encloses a greenish (Lim, 2015). (Lim, 2015). 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). Bagacay (Bambusa vulgaris) Habit: An erect plant that can grow 20 m tall and 10 cm Carbohydrates, glycosides, Used for kidney diseases, fever, diarrhea, in diameter. Stems are yellowish or yellowish-green saponins, alkaloids, inflammations, measles, gonorrhea, Leaves: Leaves have spikelets that are oblong and flavonoids, phenolics and tuberculosis, wounds, and ulcers clustered near the branches of inflorescence, lanceolate, tannins, phytosterols, and (Owolabi and Lajide, 2015). 35 cm long and 4 cm wide triterpenoids (Fitri et al., Flowers: Light-brown, rare 2020). Fruits: Rare or absent due to irregular meiosis Seeds: Produces in large amount in 1-3 years and eventually dies (Zheng et al., 2020). 175 Abastillas et al. Karupi (alpinia sp.) Habit: A perennial herb with stems that are 1-2 m tall and Tannins, glycosides, Used for fungal skin disorders, 2.5 cm in diameter. phenols, carbohydrates, antirheumatic, dyspepsia, ulcers, Leaves: Oblong-lanceolate, aromatic, root tubers monoterpenes, sterols, bronchitis, diabetes, obesity, Flowers: The flower has a hollow toothed calyx that often sesquiterpenes, and inflammations, diuretic, chest pains, sore split on one side flavonoids such as throat, and kidney diseases Fruits: Round, red berries quercetin, kaempferol, (Chouni and Paul, 2018). Seeds: obtusely angular alpinin, and galangin (Chouni and Paul, 2018) (Chouni and Paul, 2018) Phytochemical components of B. Vulgaris and ethanolic extracts of B. vulgaris and Alpinia sp. Alpinia sp. against A. hydrophila. Turbidity was not observed in Table 2 shows the phytochemical compounds present all extracts at concentrations of 0.5g ml-1 and 1g ml-1, in both karupi (Alpinia sp.) and bagacay (Bambusa indicating that bacterial growth was inhibited. Thus, vulgaris). Both plants were positive for alkaloids, the lowest concentration of extracts that can inhibit carbohydrates, glycosides, phytosterols, flavonoids the growth of bacteria is 0.5 g ml-1. and phenols and tannins. MBC of various extracts against A. hydrophila Minimum Inhibitory Concentration (MIC) and Absence in the growth of A. hydrophila was observed Minimum Bactericidal Concentrations (MBC) of in all extracts at concentrations of 0.5g ml-1 and 1g ml- various plant extracts 1. MIC of various extracts against A. hydrophila can inhibit the growth of the bacteria is 0.5g/ml Table 3 shows the result of MIC of aqueous and (Table 4). Thus, the lowest concentration of the extracts that 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 MBC of various extracts against V. harveyi The MIC of the aqueous and ethanolic extracts of B. The MBC of the ethanolic extract of B. vulgaris and vulgaris as well as the aqueous extract of Alpinia sp. aqueous extract of Alpinia sp. against V. harveyi was On the other hand, 0.25g ml-1. However, it was at 0.5 g ml-1 for the the MIC of the ethanolic extracts of Alpinia sp. is 0.5g aqueous extract of B. vulgaris and ethanolic extract of ml-1 (Table 5). Alpinia sp. (Table 6). against V. harveyi is 0.25g ml-1. Table 3. Minimum Inhibitory Concentrations (MIC) of aqueous (aq.) and ethanolic (eth.) extracts against A. hydrophila. Extract A. B. vulgaris (aq.) B. B. vulgaris (eth.) C. Alpinia sp. (aq.) D. Alpinia sp. (eth.) 1g ml -1 - 0.5g ml -1 - Concentration 0.25g ml -1 + + + + 0.125g ml -1 + + + + Positive (+): Turbidity indicating growth; Negative (–): No turbidity indicating absence of growth. MIC of various extracts against B. albus in all concentrations for Bacillus albus (Table 7). The turbidity was present in all concentrations of ethanolic and aqueous extracts indicating positive MBC of various extracts against B. albus results for bacterial growth. There was no inhibition The presence of bacterial growth was observed in all 176 Abastillas et al. extracts. Thus, at all concentrations, there was no septica.); Talus (Homalomena rubescens (Roxb.) inhibition of bacterial growth (Table 8). Kunth); Palochina Sinaw-sinaw (Sena (Peperomia alata); Badyang pellucida); (Alocasia Discussion macrorrhizos); Bagacay (Bambusa vulgaris) and A total of ten (10) medicinal plants including Karupi (Alpinia sp.) were identified from the local Hagonoy (Chromolaena odorata); Pitogo (Cycas community. The majority of these plants were rumphii); Adgao; (Premna odorata); Labnog (Ficus 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 - A. B. vulgaris (aq.) B. B. vulgaris (eth.) C. Alpinia sp. (aq.) D. Alpinia sp. (eth.) 0.5g ml -1 - 0.25g ml -1 + + + + 0.125g ml -1 + + + + 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 - A. B. vulgaris (aq.) B. B. vulgaris (eth.) C. Alpinia sp. (aq.) D. Alpinia sp. (eth.) 0.5g ml -1 - 0.25g ml -1 + 0.125g ml -1 + + + + Positive (+): Turbidity indicating growth; Negative (–): No turbidity indicating absence of growth. The morphology of the leaves, flowers, fruits, and 2018; Haryanti et al., 2021; Van et al., 2021). seeds varies among the ten plants identified. Phytochemicals such as flavonoids, triterpenoids, Furthermore, C. odorata, F. septica, P. pellucida, S. tannins, alata, A. macrorrhizos, and B. vulgaris have are alkaloids, anthraquinone, saponins, phytosterols, and phenolics have been found in these regarded as plants (Vital et al., 2010; Vijayaraghavan et al., 2017; inflammatory agent and has been used for treating Chouni and Paul, 2018). In addition, C. odorata, F. many illnesses including headache, stomachache, septica, and H. rubescens have also been identified to urinary tract infections, tuberculosis and diabetes have antibacterial, antifungal and anticancer effects (Ooi et al., 2012; Vijayaraghavan et al., 2017, Jangad (Vijayaraghavan et al., 2017; Jangad and Licardo, and Licardo, diuretic, 2018; analgesic, Haryanti laxative, et al., anti- 2021). Table 6. Minimum Bactericidal Concentrations (MBC) of aqueous (aq.) and ethanolic (eth.) extracts against V. harveyi. Extract A. B. vulgaris (aq.) B. B. vulgaris (eth.) C. Alpinia sp. (aq.) D. Alpinia sp. (eth.) Concentration 1g ml -1 - 0.5g ml -1 - Positive (+): Indicating growth; Negative (–): Indicating absence of growth. 177 Abastillas et al. 0.25g ml -1 + + 0.125g ml -1 + + + + The phytochemical analysis of B. vulgaris and Alpinia with the use of established screening methods, sp. revealed the presence of alkaloids, carbohydrates, discovered glycosides, phytosterols, flavonoids and phenols and alkaloids, flavonoids, steroids, and quinones. In the tannins. The presence of phytochemicals has also study of Fitri et al. (2020), B. vulgaris has been been found in the same genus of Alpinia sp. which is reported Alpinia officinarum. It was stressed in the study by saponins, Balamurugan et al. (2019) that the alcoholic extracts tannins, the to presence contain alkaloids, of phenols, carbohydrates, flavonoids, phytosterols, tannins, glycosides, phenolics and and triterpenoids. Table 7. Minimum Inhibitory Concentrations (MIC) 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 (+): Turbidity indicating growth; Negative (–): No turbidity indicating absence of growth. Both the minimum inhibitory concentration (MIC) vulgaris and minimum bactericidal concentration (MBC) concentrations 31.25 - 125 g ml-1 against several results of B. vulgaris and Alpinia sp. against A. gastro-intestinal microorganisms such as E.coli, K. hydrophila, a Gram-negative bacterium, indicated pneumoniae, P. mirabilis, and S. typhi in the study of minimum inhibition of all extracts at 0.5g ml-1. In V. showed MIC values ranging at Ogu et al. (2011). harveyi, all extracts except ethanolic Alpinia sp. exhibited a MIC at 0.25 g ml-1. The MBC for the The phytochemical compounds present in Alpinia sp., ethanolic extract of B. vulgaris and aqueous extract of such as tannins, alkaloids, flavonoids and saponins, ml-1, whereas the aqueous have been detected to possess antibacterial activity extract of B. vulgaris and ethanolic extract of Alpinia (Rini et al. 2018). The same phytochemicals were Alpinia sp. was 0.25g sp. was at 0.5g ml-1. This clearly demonstrates that B. found to be present in the study which largely vulgaris and Alpinia sp. are effective antibacterial contributes to the potent antibacterial activity of the agents against these two gram-negative bacteria, A. aqueous and ethanolic extracts of Alpinia sp. against hydrophila and V. harveyi. A. hydrophila and V. harveyi. In the study of Oonmetta-aree et al. (2006), among the four plants Previous studies have revealed that bamboo possesses tested, the ethanolic extracts of Alpinia galanga antioxidant, anticancer and anti-microbial activities. demonstrated the highest inhibitory activity against Bioactive compounds found in B. vulgaris, a bamboo S. aureus. species, include alkaloids, flavonoids, saponins, and tannins that contribute to its antibacterial activity The findings of the study showed the minimum (Tanaka et al., 2014). In the previous findings of inhibitory concentration (MIC) of the ethanolic Zhiang et al. (2019), ethanolic extracts from leaves of extract at 0.325 mg ml-1 and the minimum B. vulgaris have been found to exert minimum bactericidal concentration (MBC) at 1.3 mg ml-1 using inhibitory effects against a variety of bacterial species, the broth diluteion method). However, in the study of namely S. aureus and B. subtilis, at a concentration of Voravuthikunchai et al. (2005), aqueous extracts of A. 20g ml-1. (Zhiang et al., 2019). Additionally, the galanga did not show any inhibitory activities against ethanolic extracts and hot aqueous extracts of B. S. aureus. 178 Abastillas et al. 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 Acknowledgment similar The to the two Gram-negative bacteria authors greatly acknowledge the support mentioned previously. B. albus is a spore-forming, provided by the Friar administrators of the University Gram-positive, facultative anaerobe bacterium. The of San Agustin and the technical assistance of the presence of turbidity in all extracts indicates laboratory staff at the Department of Biology, College bacterial growth. The reason for this is that sporeforming 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 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- not able to penetrate the thick cell wall components inflammatory of the spores. In a related study, Bacillus subtilis International Journal of Advance Research and which Innovative Ideas in Education 5, 1125-1140. is bacteria, also was a Gram-positive used as a test spore-forming organism and anti -arthritic activity. for determining the antibacterial activity of B. vulgaris Carag H, Buot Jr IE. 2017. A checklist of the orders (Zhiang et al., 2019). However, the results indicated and families of medicinal plants in the Philippines. that B. vulgaris was found to be an effective Sylvatrop, the Technical Journal of Philippine antibacterial agent at a concentration of 20g ml-1. It Ecosystems and Natural Resources 27(1&2), 49-9. 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. 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. Conclusion The results from this study showed that aqueous and ethanolic extracts from B. vulgaris and Alpinia sp. Conchou L, Cabioch L, Rodriguez LJV, Kjellberg F. 2014. Daily rhythm of mutualistic contained phytochemical components that possess pollinator activity and scent emission in Ficus potent bactericidal activities against Gram-negative septica: bacteria, A. hydrophila and V. harveyi. These two occurring pollinators and potential consequences for least-studied medicinal plants can be further explored chemical communication and facilitation of host as potential sources of bioactive substances for the speciation. PloS ONE 9(8), e103581. development of novel antibacterial drugs. https://doi.org/10.1371/journal.pone.0103581. 179 Abastillas et al. ecological differentiation between co- Cordero C, Alejandro GJ. 2021. Medicinal plants Khalid S, Shahzad A, Basharat N, Abubakar used by the indigenous Ati tribe in Tobias Fornier, M, Anwar P. 2018. Phytochemical screening and Antique, analysis of selected medicinal plants in Gujrat. Philippines. Biodiversitas Journal of Biological Diversity 22(2). Journal of Phytochemistry and Biochemistry 2(1), 1- https://doi.org/10.13057/biodiv/d220203. 3. 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. F, Chen M, Hatab SR. Antibacterial activity of Cycas rumphii Miq. leaves extracts against some tropical human pathogenic bacteria. Research Journal of Microbiology 6(10), 761. Gonelimali FD, Lin J, Miao W, Xuan J, Charles Khan AV, Ahmed QU, Khan AA, Shukla I. 2011. 2018. Antimicrobial properties and mechanism of action of some plant extracts against food pathogens and Lim TK. 2015. Alocasia macrorrhizos. Edible Medicinal and Non Medicinal Plants: Volume 9, Modified Stems, Roots, Bulbs. Springer, London. 429-442 p. spoilage microorganisms. Frontiers in Microbiology 9, 1639. Lirio SB, Macabeo AP, Paragas EM, Knorn M, https://doi.org/10.3389/fmicb.2018.01639. Kohls P, Franzblau SG, Wang Y, Aguinaldo MA. 2014. Antitubercular constituents from Premna Haryanti S, Rahmawati N, Sholikhah I, odorata Blanco. Journal of Ethnopharmacology 154 Widiyastuti Y. 2021. Ficus septica, an ecosystem (2), 471-474. keystone species induced ROS-mediated cytotoxicity https://doi.org/10.1016/j.jep.2014.04.015 in HepG2 hepatocarcinoma cells. Earth Environmental Science 905(1), 012101. Mollejon B, Mollejon C. 2019. Anti-microbial http://dx.doi.org/10.1088/1755-1315/905/1/012101. activities and phytochemical screening of the Premna odorata Blanco alagaw leaf extract. International Hill KD. 1994. The Cycas rumphii complex (Cycadaceae) in New Guinea and the western Pacific. Australian Systematic Botany 7(6), 543-567. Journal of Trend in Scientific Research and Development 3, 343-345. Nuneza O, Rodriquez B, Nsaid JG. 2021. https://doi.org/10.1071/SB9940543. Ethnobotanical survey of medicinal plants used by the Mamanwa tribe of Surigao del Norte and Agusan del Hussain W, Badshah L, Ullah M. 2018. Quantitative study of medicinal plants used by the communities Northern residing in Pakistani-Afghan Koh-e-Safaid borders. Norte, Mindanao, Philippines. Biodiversitas 22(6). https://doi.org/10.13057/biodiv/d220634. Range, Journal of Ogu GI, Madagwu EC, Eboh OJ. 2011. Ethnobiology and Ethnomedicine 14(1), 1-18. Antibacterial activities of three medicinal plants https://doi.org/10.1186/s13002-018-0229-4. against some gastro-intestinal microorganisms. International Journal of Natural and Applied Sciences Jangad AMA, Licardo ADMB. 2018. Acute and 7(3), 248-256. chronic toxicity studies of lagnob (Ficus septica Burm. F. 1768) fruit extract on albino rats (Rattus Oladeji O. 2016. The characteristics and roles of norvegicus). Doctoral dissertation, Thesis. University medicinal plants: some important medicinal plants in of San Carlos-Josef Baumgartner Learning Resource Nigeria. Natural Products: An Indian Journal 12(3), Center. Talamaban, Cebu City, Philipines. 180 Abastillas et al. 102. Oladeji O, Adelowo F, Oluyori A, Bankole D. Sofowora A, Ogunbodede E, Onayade A. 2013. 2020. Ethnobotanical Description and Biological The role and place of medicinal plants in the Activities strategies for disease prevention. African Journal of of Senna alata. Evidence-Based Complementary and Alternative Medicine 2020, 2580259. Traditional, Complementary, and Alternative Medicines 10(5), 210–229. https://doi.org/10.1155/2020/2580259. http://dx.doi.org/10.4314/ajtcam.v10i5.2. Ooi DJ, Iqbal S, Ismail M. 2012. Proximate composition, nutritional attributes and mineral Talib WH, Mahasneh AM. 2010. Anti-microbial, composition cytotoxicity of Peperomia pellucida L. and phytochemical screening of (Ketumpangan Air) grown in Malaysia. Molecules Jordanian plants used in traditional medicine. (Basel, Switzerland) 17(9), 11139–11145. Molecules 15(3), 1811-1824. https://doi.org/10.3390/molecules170911139. https://doi.org/10.3390/molecules15031811. Oonmetta-aree J, Suzuki T, Gasaluck P, Eumkeb G. 2006. Antimicrobial properties and action of galangal (Alpinia galanga Linn.) on Staphylococcus aureus. Lebensmittel-Wissenschaft Tanaka A, Zhu Q, Tan H, Horiba H, Ohnuki K, Mori Y, Yamauchi R, Ishikawa H, Iwamoto A, Kawahara H, Shimizu 2014. K. Biological Technologies-Food Science and Technology 39(10), activities and phytochemical profiles of extracts from 1214-1220. different https://doi.org/10.1016/j.lwt.2005.06.015. pubescens). Molecules 19(6), 8238–8260. parts of bamboo (Phyllostachys https://doi.org/10.3390/molecules19068238 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. 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. Rini C, Rohmah J, Widyaningrum L. 2018. The http://dx.doi.org/10.15414/jmbfs.2020.10.2.201-204. antibacterial activity test galanga (Alpinia galangal) on the growth of bacteria Bacillus subtilis and Vijayaraghavan K, Rajkumar J, Bukhari SN, Escherichia coli. IOP Conference Series: Materials Al‑ ‑Sayed B, Seyed MA. 2017. Chromolaena 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, 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. Chromolaena odorata. Pharmacognosy Reviews 11 (21), 35–38. Voravuthikunchai http://dx.doi.org/10.4103/phrev.phrev_53_16. Subhadhirasakul SP, S. Phongpaichit 2005. Evaluation S, of antibacterial activities of medicinal plants widely used Shaikh J, Patil M. 2020. Qualitative tests for preliminary phytochemical screening: an overview. International Journal of Chemical Study 8(2), 603608. 181 Abastillas et al. among AIDS patients in Thailand. Pharmaceutical Biology 43(8), 701-706. https://doi.org/10.1080/13880200500385194. Vital P, Velasco R, Demigillo J, Rivera W. Zheng X, Lin S, Fu H, Wan Y, Ding Y. 2020. The 2010. bamboo flowering cycle sheds light on flowering Antimicrobial activity, cytotoxicity and phytochemical screening of Ficus septica Burm and diversity. Frontiers in Plant Sciences 11, 381. Sterculia foetida L. leaf extracts. Journal of Medicinal https://doi.org/10.3389/fpls.2020.00381. Plants Research 4(1), 58-63. http://dx.doi.org/10.5897/JMPR09.400. Zhiang K, Sari R, Apridamayanti P. 2019. Youssef FS, Ovidi E, Musayeib NMA, Ashour Bambusa vulgaris leaves extract against skin and ML. 2021. digestive diseases bacteria. Kartika: Jurnal Ilmiah Minimum inhibitory concentration etermination of Morphology, anatomy and secondary metabolites investigations of Premna odorata Blanco Farmasi 7(1), 1-5. and evaluation of its anti-tuberculosis activity using http://dx.doi.org/10.26874/kjif.v7i1.158. in vitro and in silico studies. Plants 10(9), 1953. https://doi.org/10.3390/plants10091953. 182 Abastillas et al.