Research Paper Antimicrobial Activity of Crude Oil Extracts from Fruits Of

Research Paper

Antimicrobial activity of crude oil extracts from fruits of Cynometra ramiflora Linn and Pandanus conoideus Lamk from Papua New Guinea against ten pathogenic micro-organisms

Accepted 17th October, 2019

ABSTRACT

Plant-derived medicines are increasingly sought as alternatives to synthetic drugs. Adverse side-effects of antibiotics and resistance to certain pathogens are the reasons. This study investigated crude oil extracts of the fruits/seed kernels of Cynometra ramiflora and Pandanus conoideus and examined their efficacies against ten pathogens. The oils were assayed using disc (agar) diffusion technique against 8 bacteria, yeast and protozoa. The efficacies of the micro-organisms were evaluated by inhibition zones (IZs) and the potency of the oils were compared against antibiotics Gentamycin (10 µg) and Erythromycin (10 µg). Initially, 250 ml of crude oil was produced from 3.0 kg of pounded seeds of C. ramiflora while 500 ml of oil was extracted from 1.5 kg pericarps of P. conoideus using Soxhlet extractor and hot-water technique, respectively. The oils were designed in suspensions as: C. ramiflora 100% pure oil (CR), P. conoideus 100% pure oil (PC1), mixture of P. conoideus oil (10%) and palm oil (90%) (PC2) and mixture of P. conoideus oil (20%) and palm oil (80%) (PC3). Under sterile conditions, subcultured microbes (100 µL) were transferred onto agar plates. Then 40 paper discs (6 mm diameter) were impregnated with 25 µL of each oil suspensions. The impregnated discs were transferred to seeded media. Ten replicates were prepared for each oil suspensions and incubated at 30-37C for 24-48 h. After incubation, the IZs were measured for each replicate twice and an average reading Rossie Wagi1 and Benson K. was noted. As per the results, pure oil of C. ramiflora indicated minimal efficacy (IZ 6 mm) 2* Gusamo against all test pathogens except for Micrococcus luteus with nil bioactivity. A maximum

1Forestry Department, PNG antibacterial activity was observed against Salmonella typhimurium (IZ 17 mm) equivalent University of Technology, Lae, to antibiotic Gentamycin (IZ 17 mm) but less effective than Erythromycin (IZ 30 mm). Morobe Province, Papua New Alternatively, the oils of P. conoideus showed nil to low efficacy against ten pathogens. PC1 Guinea. demonstrated minimal antibacterial activity (IZ 6 mm) against Bacillus cereus, Escherichia 2Bulolo University College, P.O. Box coli and Neisseria gonorrhoeae while it was ineffective against other seven micro-organisms. 92, Bulolo, Morobe Province, Papua PC2 showed no potency against nine micro-organisms except for N. gonorrhoeae with New Guinea. minimal IZ (6 mm). PC3 was ineffective (IZ 0 mm) against ten micro-organisms screened.

*Corresponding author. E-mail: [email protected]. Tel: +675 Key words: crude oil extracts, Cynometra ramiflora, Pandanus conoideus, microbial 474 5226. susceptibility screening, pathogenic micro-organisms, inhibition zone, Papua New Guinea

INTRODUCTION

Plants continue to provide raw materials for many benefits. About 25% of prescribed drugs used globally are modern herbal and pharmaceutical drugs to treat various sourced from plants (Rates, 2001) and ca. 90% of all ailments and diseases as well as offer other health human diseases (bacterial infections, cancer, and

Academia Journal of Medicinal Plants; Wagi and Gusamo. 237

immunological disorders) are treated with natural Euphorbia geniculata to have antibacterial potency. It is products (Newman and Gragg, 2007). Particularly, in the anticipated that PNG’s high biodiversity forest ecosystem developing world, traditional medicine (herbs) is an holds medicinal plants with huge potential for important part of livelihoods used by generations. FAO conventional drugs. (2004) reported that 80% of the population in developing The plant species of interest in this research are crude countries depend on medicinal plants for primary health oil extracts from the fruits of Cynometra ramiflora Linn care needs. In fact, modern drugs evolved from folk and and Pandanus conoideus Lamk. C. ramiflora (Fabaceae) is traditional medicines (Morsy, 2014). Jain et al. (2019) a small to medium-sized tree (Afrin et al., 2016) found in added that plant-derived drugs were part of human tidal and black-mangrove forests of the lowland forest up evolution and healthcare for thousands of years. The use to 400 m above sea level (Useful Tropical Plants Website, of Pongamia pinnata L and Terminalia chebula has a long 2018). The tree is distributed in the tropical regions of history with Ayurveda, Siddha, and Unani systems of PNG, Indonesia (Asia) and Australia (Cooper, 2015). Many traditional medicine in India (Muthu et al., 2006; Bai, investigations of the species in Asia involved isolation of 2019). Morinda citrifolia is used for millennia as a panacea ingredients from the leaves and stem barks, for wide range of ailments/afflictions in the Pacific phytochemical analysis and test for microbial efficacy (Sakulas et al., 2010). The main reasons for seeking herbal (Afjalus et al., 2013). For instance, the authors evaluated medicine are easy accessibility, affordability, limited bark extract for neuropharmacological, antibacterial and supply of synthetic drugs in hospitals/pharmacies and the antinociceptive effects in Bangladesh. Also, a leaf extract trust for the herbs. In addition, many are forced to seek synthesized iron oxide nanoparticles indicated effective herbal medicines due to adverse side-effects of many antibacterial activity and stressed for developing synthetic drugs and resistance of certain pathogens to antibacterial drug (Grois et al., 2016). Additionally, a leaf antibiotics (Jain et al., 2019). Thus, numerous researches extract tried in Indonesia showed antiviral potency were pursued into medicinal plants for their against Dengue virus (Meutia et al., 2017). However, very phytochemistry and bioactivity in many parts of the little work has been reported on seed (kernel) extracts world. For instance, >3,000 plant species were reported and their bioactivity. Merca et al. (1989) isolated a lectin to have anti-cancer properties (Graham et al., 2000; Uddin (highly acidic protein) from seeds while oil extracts from et al., 2009) and >500 medicinal plants were discovered seeds were used to make lotion for skin diseases (Siraj et in Bangladesh (Uddin et al., 2009). Shrestha et al. (2015) al., 2013; Afrin et al., 2016). recorded 33 medicinal plants in Nepal. Also, Indian On the other hand, P. conoideus (Pandanaceae) is a studies found seed extracts of Parkia javanica to have monocot endemic to the island of New Guinea (Bourke, anticancer effect (Khangemban et al., 2018) while fruit 2005; Rohman and Windarsih, 2018). It grows in tropical extracts of Algerian grown Fraxinus excelsior have environments from lowlands up to 2000 m altitude antioxidant and antibacterial effects (Amamra et al., (Stone, 1992) and commonly found near creeks. P. 2018). conoideus bears an edible red/orange and yellow fruit In Papua New Guinea (PNG), traditional herbs for (pericarp); thus, it is commonly called red fruit medicine provide primary health care needs for the bulk (Indonesia) and marita (PNG). In Indonesia, red fruit has of the population. About 70% of population (from 8.5 been used by Papua people as a natural remedy for million) lives in rural communities located in variety of diseases (Wismandanu et al., 2016; Achadiyani rugged/mountainous or inundated country where et al., 2016). In PNG, P. conoideus is cultivated in gardens accessibility to infrastructures (road, transport, for edible oil (Gusamo and Jimbudo, 2015) and culturally communication and health services) is often impossible. it is considered as a delicacy in the highlands region. PNG, being a developing country, tropical Numerous studies on phytochemistry (characterization) ailments/diseases are prevalent and many rural and bioactivities of the oil extract are well documented communities rely on plant-derived medicines to treat based on Indonesian grown material (Rohman and these diseases. The first case study into PNG medicinal Windarsih, 2018). Chemical analysis of the oil extract has plants began in 1977 by Holdsworth. According to high level of monounsaturated fatty acid (oleic acid), β- Woodley (1991), 127 medicinal plants belonging to 71 carotene, β-cryptoxanthin, α- tocopherol, phenolic families were identified. Subsequently, Wau et al. (2010) compound and flavanoid (Mun’im et al., 2006; Khie et al., reported antimicrobial activity of the fruit latex of Ficus 2009; Murtiningrum et al., 2012; Rohman et al., 2012; botryocarpa and Zure et al. (2010) identified Salazar, 2018). The following health benefits were immunostimulant activity from the phytochemicals highlighted: strengthens body’s immune system and (alkaloids and isoflavanoids) of Piper bolanicum. Recently, inhibits virus that triggers HIV/AIDS, heals/prevents Timi et al. (2018) found isolate of perennial plant spread of cancer, lowers blood pressure and prevents

Academia Journal of Medicinal Plants; Wagi and Gusamo. 238

stroke, provides sufficient insulin and prevents diabetes, shelled) with a knife. More than 300 seed kernels were prevents osteoporosis and improves brain functions (Khie weighed for green (initial) weight and dried at room et al., 2009; Rohman et al., 2012; Mun’im et al., 2006; temperature for 2 days. The pre-dried seed kernels were Wismandanu et al., 2016; Salazar, 2018). Additionally, the re-weighed and milled (pounded) into 3000 g powdery oil treats hypertension, heart disease (arteriosclerosis) form. For P. conoideus, three ripe fruits (pericarps) were (Budi and Paimin, 2004); reduces obesity and increases collected from Kimagl village, Kerowagi District, Simbu lipolytic activity in adipose tissue, prevents age-related Province. The pericarps were stored in a cool room cognitive decline and dementia (Asensio et al., 2008) and (refrigerator) for 10 days to prevent biodegradation. malaria (Limbongan and Malik, 2009). Next, Indrawati During storage, mold fungi growths were observed prior (2016) reported minimal to maximum antibacterial to extraction and experimentation. activity (inhibition zones 8.28-32.0 mm) against common bacteria. Further, an acute toxicity study in rats indicated non-toxicity and categorized as Category 5 GHS (Globally Aqueous extract of crude oils Harmonized System for Chemical Classification Substances and Mixtures) (Wismandanu et al., 2016). A Soxhlet extractor was used to extract crude oil of C. Furthermore, Rohman and Windarsih (2018) reported ramiflora. A wad of cotton wool was plugged in the hole of that due to many functional health benefits, the oil is the extraction unit. The 3000 g ground sample was placed highly-priced (10-15 times) than other common oils (e.g. in the extraction unit. A clean/dried round-bottom flask palm, corn, soybean, and canola) on the Indonesian was weighed and attached to the extraction unit. The market. extraction unit was filled with hexane (3730-2 SL. Class 3, Disk (agar) diffusion is a standard method widely used Minimum Assay 98.0%, Banksia Scientific Co., Australia) for assaying plant extracts for antimicrobial activity to siphon once and half-filled with hexane again. The (Perez et al., 1990) under a controlled laboratory condenser unit was attached and heated on the electric condition. This test involves isolation of plant’s heating mantle and heat was regulated to permit phytochemicals and screening the active ingredients condensation of 5-10 drops per second. The extraction against target micro-organisms for efficacy. The growth process was allowed for 8 h. The flask was then removed and proliferation, and the degree of the ingredients to and the hexane was distilled to separate the oil. Finally, 20 inhibit the growth of the organisms are evaluated (Bauer ml of oil was retained in the flask. et al., 1966). According to Prakash (2009) and Jacob et al. On the other hand, hot water extraction technique was (2016), the diameter of inhibition zones of micro- used to extract oil from P. conoideus fruits. Firstly, the organisms during microbial screening test are fruits were washed with water to remove foreign matter. rated/classified as maximum (10-20 mm), moderate (10- The fruits were weighed (2000 g), de-cored, chopped with 15 mm) and minimal to no antimicrobial activities (<10 the pericarps intact (1500 g) and boiled in water (1:1 mm). ratio) until tender. The boiled pericarps were removed This study aimed to extract crude oils of Cynometra and cooled at ambient temperature. The same water used ramiflora and Pandanus conoideus, and screened against for boiling was added (1:1 ratio) to the pericarps and strains of pathogenic micro-organisms (3 Gram-positive pulped (macerated) by squeezing with gloved-hands. The and 5 Gram-negative bacteria, 1 species each of protozoa seeds were discarded and a thick creamy substance and yeast). The antimicrobial susceptibilities (efficacies) (puree) was collected. The puree was filtered to remove of the oil extracts against the pathogens were ascertained any foreign particles, heated for a few minutes and by measuring inhibition zones using disc diffusion allowed the concentrate to cool overnight. The creamy method. Fat content analysis, characterization, and concentrate was then separated into a layer of water and phytochemistry of the fats were not part of the thick creamy oil. The creamy oil was dispensed and investigation. centrifuged at 3000 rpm for 30 min. The top layer oil was siphoned off as crude oil producing 150 ml while the rest was a puree. MATERIALS AND METHODS

Sampling and preparation of fruit samples Mixing oil extracts with palm oil as carrier solvent

More than 300 mature (ripe) fruits were collected from The oil extract of C. ramiflora was pure (100%) oil (CR) two trees of C. ramiflora growing in Lae, PNG. The hard intended to use in microbial test without mixing with coverings of fruits of C. ramiflora were removed (de- palm oil as a carrier solvent. Except for P. conoideus, the

Academia Journal of Medicinal Plants; Wagi and Gusamo. 239

Table 1: Strains of pathogenic micro-organisms.

Strains of micro-organisms Species Bacteria (gram-positive) Bacillus cereus, Micrococcus luteus, Staphylococcus aureus Bacteria (gram-negative) Escherichia coli, Klebsiella pneumoniae, Neisseria gonorrhoeae, Proteus vulgaris, Salmonella typhimurium Protozoa Trichomonas vaginalis Yeast Candida albicans

oil extracts were mixed with palm oil. The crude oils were needle, which was held over the flame to red hot mixed in three 60 ml bottles and the composition of the occasionally before use. The discs were pressed gently to mixtures were: (a) 100% pure P. conoideus oil (PC1), (b) ensure firm contact with the seeded media. The yeast 10% P. conoideus oil to 90% palm oil (PC2) and (c) 20% P. (Candida albicans) and protozoa (Trichomonas vaginalis) conoideus oil to 80% palm oil (PC3). were placed in an incubator at 30C whilst the bacteria were incubated in a separate incubator at 37C. The incubation of microbes took 24 to 48 h to facilitate Bioassay: Preparation of micro-organism, assay plates maximum growth. After incubation, the zone of inhibition and paper discs (mm) was measured for each replicate using a ruler. Each measurement of the inhibition zone was taken twice and Pure cultures of ten micro-organisms (Table 1) were an average reading was recorded (Table 2). prepared in bottles and stored in the fridge. The ten In addition, two synthetic antibiotics weighing 10 µg indicator micro-organisms selected and screened were each of Gentamycin and Erythromycin were used as pathogens responsible for inducing various diseases in standard/reference substances in the experiment. The humans and animals. preparation and test of antibiotics against the micro- Nutrient agar (11.3 g) was added in distilled water (400 organisms followed the same procedure described above. ml) and sterilized in an autoclave at 115C for 20 min. The bioactivities exhibited by two antibiotics were After sterilization, nutrient agar (15-20 ml) was compared with that of crude oil extracts (CR, PC1, PC2, dispensed into ten sterile plastic Petri-discs (standard and PC3) of the two test species (Table 2). size 100 mm internal diameter). The Petri-discs (with agar) were left to cool and solidify at room temperature prior to bioassay. Paper discs (6 mm diameter) were cut RESULTS AND DISCUSSION from Matricel filter paper (1 mm thick) with a paper-hole puncher and were sterilized. Different efficacy levels of the oil extracts of two test species as well as two reference antibiotics on the ten indicator micro-organisms are shown in Table 2. The Microbial test bioactivity displayed by the ten micro-organisms was indicated by clear zones of inhibition (IZ) in millimeter A standard disc (agar) diffusion technique was employed (mm). In this case, the higher the IZ, the higher the to conduct the microbial susceptibility screening test efficacy of the oils against the growth of micro-organisms. (Bauer et al., 1966). Onto the ten prepared agar plates, According to the results (Table 2), oil extracts (CR, PC1, 100 µL of ten subcultured microbes were transferred with PC2 and PC3) of the fruits of two test species a sterile micropipette and spread evenly using a glass demonstrated variability in their antimicrobial activity hockey rod under sterile conditions. The glass hockey rod against the ten micro-organisms screened. The slightly dipped in ethanol solution (99%) and heated over antimicrobial activities were determined by their IZs and the Bunsen burner flame for a few seconds (2-5 s) and rated according to descriptions provided by Prakash removed from the flame to cool. The microbes were (2009) and Jacob et al. (2016). The IZs (efficacies) of the streaked evenly over the Petri-discs with glass hockey oil extracts were then compared with standard rod. Using a micropipette, 40 paper discs were (synthetic) antibiotics (Gentamycin (GM) and impregnated with 25 µL suspensions of CR, PC1, PC2, and Erythromycin (EM)). The antibiotics (GM and EM) PC3 and dried. A total of 10 replicates per oil suspension indicated high efficacies against the micro-organisms. For were prepared. The impregnated discs were then instance, the IZ of GM and EM ranged between 15-24 and transferred to seeded media using a sterilized stabbing 6-30 mm respectively. In particular, EM revealed minimal

Academia Journal of Medicinal Plants; Wagi and Gusamo. 240

Table 2: Antimicrobial activity of crude oils of fruits of C. ramiflora and P. conoideus and reference substances against ten micro-organisms.

Inhibition Zone (mm) Micro-organisms CR PC1 PC2 PC3 GM EM Bacillus cereus (G+) 6 6 0 0 17 22 Micrococcus luteus (G+) 0 0 0 0 17 21 Staphylococcus aureus (G+) 6 0 0 0 24 23 Escherichia coli (G-) 6 6 0 0 19 28 Klebsiella pneumoniae (G-) 6 0 0 0 16 15 Neisserria gonorrhoeae (G-) 6 6 6 0 17 25 Proteus vulgaris (G-) 6 0 0 0 21 6 Salmonella typhimurium (G-) 17 0 0 0 17 30 Candida albicans (Y) 6 0 0 0 15 6 Trichomonas vaginalis (Pz) 6 0 0 0 18 20

G+: gram positive bacteria, G-: gram negative bacteria, Y: yeast, Pz: protozoa, CR: pure C. ramiflora oil, PC1: pure P. conoideus oil, PC2: mixture of P. conoideus oil and palm oil, PC3: mixture of P. conoideus oil and palm oil, GM: Gentamycin, EM: Erythromycin.

IZs (6 mm) for Proteus vulgaris (G-) and yeast (C. albicans) typhimurium (e.g. Gastroenteritis (gut inflammation), whilst it was significantly effective against Escherichia coli Diarrhoea, Vomiting, Fever, Abdominal cramps) (G-) and Salmonella typhimurium (G-) with IZs (28 and 30 (https://www.yourgenome.org/facts/what-is-salmonella, mm) respectively. On the other hand, GM proved 2018). moderate to high efficacy against all micro-organisms screened. Bioactivity of Pandanus conoideus oil extracts

Bioactivity of Cynometra ramiflora oil extracts The oil extracts of P. conoideus in three different forms (PC1, PC2 and PC3) indicated nil to minimal antimicrobial The oil extract of C. ramiflora (CR 100%) showed nil to activity against ten micro-organisms tested. A minimal minimal efficacy/potency (IZ 0-6 mm) for ten strains of antibacterial activity with IZ (6 mm) was observed for pathogens screened except for Salmonella typhimurium PC1 (100%) when screened against three bacteria (G-). It was ineffective against Micrococcus luteus (G+) (IZ Bacillus cereus (G+), E. coli (G-) and Neisserria 0 mm) while a minimum antimicrobial activity (IZ 6 mm) gonorrhoeae (G-) whilst it was ineffective against seven was observed for nine micro-organisms tested apart from other micro-organisms (IZ 0 mm). For the case of PC2 bacterium S. typhimurium (G-). Additionally, it was noted (mixtures of P. conoideus (20%) and palm oil (80%)), it that CR oil indicated antimicrobial activity similar to that showed minimal antibacterial activity with IZ (6 mm) of antibiotic EM (IZ 6 mm) against bacterium P. vulgaris against bacterium N. gonorrhoeae (G-). The oil was (G-) and yeast C. albicans. Further, an interesting ineffective against nine other micro-organisms (IZ 0 mm). observation was made for CR oil, which exhibited a Moreover, PC3 (mixtures of P. conoideus (10%) and palm maximum efficacy (IZ 17 mm) against bacterium S. oil (90%)) were significantly ineffective against ten typhimurium (G-). Comparatively, the antibacterial strains of micro-organisms screened with IZ (0 mm). The activity demonstrated by CR oil was comparable results of this research contradict the findings of (equivalent) to IZs of antibiotics GM (17 mm) and EM (30 Indrawati (2016), who reported minimal to maximum mm) for the same bacterium. Similar observations antibacterial activity. The microbial efficacy of the P. (effective antibacterial activities) were reported by conoideus oil was nil to minimal and incomparable to Afjalus et al. (2013) and Grois et al. (2016) from the bark standard antibiotics used in this study. Also, it was and leaf extracts of C. ramiflora, respectively. The finding observed that increasing the amount of palm oil (%) as a of this case study indicates that oil extract of seed kernels carrier solvent in the mixture with P. conoideus oil of C. ramiflora holds potential for developing an decreased the efficacy against the micro-organisms. antibacterial drug to treat diseases inflicted by S. Therefore, the results of this case study implied that oil

Academia Journal of Medicinal Plants; Wagi and Gusamo. 241

extracts of P. conoideus have poor antimicrobial University of Melbourne, Australia, read the paper. phytochemicals. The low efficacy of P. conoideus oil could be due to the onset of rancidity. There was a delay (10 days) between harvesting of the fruits and REFERENCES experimentation. Thus, the active ingredients of the fruits were suggested to have undergone partial Achadiyani, Septiani L, Fareid A, Bolly HMB, Dikdik K (2016). Role of the red fruit (Pandanus conoideus LAM) Ethyl Acetate fraction on the oxidation/hydrolysis upon exposure to the environment induction of apoptosis vs. downregulation of survival signalling and/or biodegraded via the actions of mould fungi. Also, pathways in cervical cancer cells. Eur. J. Med. Plants. 13(2):1-9. the low efficacy could be attributed to thermal Afjalus S Md, Malik S, Emrul H, Sanjana S, Farjana Y (2013). Evaluation of degradation of volatile compounds during hot water neuropharmacological, antibacterial and antinociceptive activity of methanolic extract of the bark of Cynometra ramiflora Linn. extract/heating extraction (Das et al., 2010). (Leguminosae). Int. J. Res. Ayurveda Pharm. 4(2):192-197. Afrin S, Pervin R, Sabrin F, Rony SR, Sohrab Md H, Islam Md E, Islam KD, Billah Md M (2016). In Vitro antioxidant activity, antimicrobial and CONCLUSION preliminary cytotoxic activity of Cynometra ramiflora – A Mangrove Plant. J. Microbiol. Biotechnol. Food Sci. 6(2): 844-850. Amamra S, Charef N, Arrar L, Belhaddad O, Khennouf S, Zaim K, Baghiani The crude oil extracts of C. ramiflora showed minimal A (2018). Phenolic content, antioxidant and antibacterial effects of antimicrobial activities against nine micro-organisms fruit extracts of Algerian ash, Fraxinus excelsior. Indian J. Pharm. Sci. screened while a high (maximum IZ) antibacterial efficacy 80(6):1021-1028. Asensio L, Gonzalez I, Garcia T, Martin R (2008). Review: Determination was observed against Salmonella typhimurium (G-). The IZ of food authenticity by enzyme-linked immunosorbent assay (ELISA). (mm) demonstrated by C. ramiflora against the same Food Control. 19(1): 1-8. https://www.yourgenome.org/facts/what- bacterium was comparable (equivalent) to synthetic is-salmonella, 2018. antibiotics tried in the experiment. The finding indicates Bai DD (2019). Authentication of seven Indian traditional types of the herbal drug terminalia chebula with scientific updates – An advanced the potential of the oils for developing an antibacterial insight. Int. J. Pharm. Sci. Res.10(6): 2670-2687. drug for Salmonella typhimurium (G-) infections. Further Bauer AW, Kirby WM, Sherris JC, Truck M (1966). Antibiotic research on the analysis of phytochemical constituents susceptibility testing by a standardized single disc method. Am. J. Clin. (including acute toxicity test) of oil extract of the seeds for Pathol. 45(4): 493-496. Bourke RM (2005). Indigenous fruit in Papua New Guinea. Report of pharmaceutical and therapeutical purposes is Department of Human Geography. Australia: The Australian recommended. University. pp. 1-11 In contrast, oil extracts of the pericarps of P. conoideus Budi M, Paimin FR (2004). Red fruit (Pandanus conoideus L). Jakarta: were ineffective (low to minimal antimicrobial activity) Penebar Swadaya. 3-26,47-56,67-68. Cooper WE (2015). A taxonomic revision of Cynometra L. (Fabaceae) in against all micro-organisms screened. PC1 (pure oil Australia with a new species from the Wet Tropics of Queensland and extract) indicated minimal antibacterial activity. The a range extension to the mainland. Austrobaileya. 9(3): 393-403. finding showed that oil extract of pericarps of the P. Das K, Tiwari RKS, Shrivastava DK (2010). Techniques for evaluation of conoideus had no antimicrobial activity under the medicinal plant products as antimicrobial agents: Current methods and future trends. J. Med. Plants Res. 4(2):104-111. conditions used. To validate the findings of Indrawati FAO (2004). Trade in Medicinal Plants. Economic & Social Department. (2016), a repeat bioassay test using fresh P. conoideus FAO of the UN. Rome, Italy. fruits immediately after harvest to avoid rancidity is Graham JG, Quinn ML, Fabricant DS, Farnsworth NR (2000). Plants used recommended. Also, the use of polar (methanol) and non- against cancer-an extension of the work of Jonathan Hartwell. J. Ethnopharmacol. 73(3): 347-377. polar (hexane) solvents for crude oil extraction instead of Grois S, Selvaraj R, Varadavenkatesan T, Vinayagam R (2016). Structural hot water/heat extraction to exclude thermal degradation characterization, antibacterial and catalytic effect of iron oxide of active (volatile) compounds is desirable. nanoparticles synthesized using the leaf extract of Cynometra ramiflora. J. Mol. Struct. 1128: 572-578. Gusamo BK, Jimbudo M (2015). Tree-borne oilseed crops: Jatropha curcas, Ricinus communis, Anacardium occidentale and some native ACKNOWLEDGEMENTS trees for oil production for Bio-energy source in Papua New Guinea. J. Agric. Environ. Sci. 4(2):113-123. Holdsworth DK (1977). Medicinal plants of Papua New Guinea. South Kind assistance was provided by Reilly Nigo, Wap Kuipa, Pacific Commission, Noumea. Nehemiah Pelis Wangiwan and Richard Laki in the Indrawati I (2016). Sensitivity of pathogenic bacteria to Buah Merah laboratory experiments, Applied Science Department, (Pandanus conoideus Lam.). AIP Conference Proceedings. 1744(1): 10.1063/1.4953502 PNG University of Technology. Mr. Artis Vinas of the Jacob DR, Vigasini N, Iyer P (2016). Antibacterial activity of mangosteen Forestry Department, PNG University of Technology (Garcinia mangostana) Pericarp. World J. Pharm. Res. 5(9):1797- identified the plant species. Dr. Anthony Harakuwe of 1802. Roots Organic Inc commented on the manuscript. While Jain C, Khatanu S, Vijayvergia R (2019). Bioactivity of secondary metabolites of various plants: A Review. Int. J. Pharm. Sci. Res. 10(2): Professor Barbara Ozarska and Dr. Benoit Belleville of the 494-504.

Academia Journal of Medicinal Plants; Wagi and Gusamo. 242

Khangembam VC, Srivastava SK, Leishangthem GD, Thakuria D (2018). Salazar E (2018). Top Six Health Benefits of Buah Merah. Retrieved from Evaluation of apoptosis Inducing ability of Parkia javanica seed Aim High Tolling Solutions Inc: http://www.aimhigh.ph/top-6-health- extract in cancer cells. Indian J. Pharm. Sci. 80(6):1069-1077. benefits-of-buah-merah-fruit/ Khie K, Andhika OA, Chakravitha M (2009). Inhibition of NF-KB pathway Shrestha P, Adhikari S, Lamichhane B, Shrestha BG (2015). Phytochemical as the therapeutic potential of red fruit (Pandanus conoideus Lamk) in Screening of the Medicinal Plants of Nepal. IOSR J. Environ. Sci. Toxicol. the treatment of inflammatory bowel disease. J. Korean Med. 9(1):69- Food Technol. 1(6):11-17. 75. Siraj AM, Malik S, Emrul H, Sanjana S, Farjana Y (2013). Evaluation of Limbongan J, Malik A (2009). Opportunity of red fruit crop (Pandanus neuropharmachological, antibacterial and antinociceptive activity of conoideus Lam.) development in Papua Province. J. Penelit. Tanam. methanolic extract of the bark of Cynometra ramiflora Linn. Ind. 28(4):134-141 (Leguminosae). Int. J. Res. Ayurveda Pharm. 4(2):192-197. Merca FE, Macalma TB, Macapinlac MP(1989). Isolation and partial Stone BC (1992) In: Verheij E.W.M & Coronel, R.E. (Edits.) Edible fruits and characterization of a lectin from Cynometra ramiflora. Philipp. J. Sci. nuts. PROSEA. 2: 240-243. Meutia Zahra Z, Dewi, Ernawati B (2017). Effects of C. ramiflora Linn Timi D, Gopalakrishnan S, Maino M (2018). Characterization and Leaf extract against dengue virus replication in vitro on Huh7it-1 Cell. antimicrobial assessment of photosynthesized silver nanoparticles using Adv. Sci. Lett. 23(7): 6834-6837. aqueous extract of Euphorbia genculata. Indian J. Sci. Technol. 11: 47. Morsy NM (2014). Phytochemical analysis of biologically active Uddin SJ, Grice ID, Tiralongo E (2009). Cytotoxic Effects of Bangladeshi constituents of medicinal plants. Main Group Chem. 13: 7-21. medicinal plant extracts. Evid. Based Complementary Altern. Med. Mun’im A, Andrajati R, Susilowati H (2006). Tumorigenesis inhibition of Article ID 578092. 7 pp. doi:10.1093/ecam/nep111 water extract of red fruit (Pandanus conoideus Lamk) on Sprague- Wau JS, Harakuwe AH, Sakulas HW, Timi R, Gubag-Sipou R, Wangiwan P Dawley rat female induced by 7.12- dimetilbenz(a)antrasen (DMBA). (2010). Studies on PNG Medicinal Plants: The efficacy of the methanol Indonesia J. Pharm. Sci. 3:153-161. extracts from the fruit latex of Ficus botryocarpa against Bacterial Muthu C, Ayyanar M, Raja N, et al (2006). Medicinal plants used by Isolates from yaws. Environment Papua New Guinea. 2(2): 7-10. traditional healers in Kancheepuram district of Tamil Nadu, India. J. Wismandanu O, Maulidya I, Indariani S, Batubara I (2016). Acute toxicity of Ethnobiol. Ethnomed. 2: 43-53. red fruits (Pandanus conoideus Lamk) oil and the hepatic enzyme level in Murtiningrum, Sarungallo ZL, Mawikere NL (2012). The exploration and rat. J. Phytopharmacology. 5(5): 176-178. diversity of red fruit (Pandanus conoideus L) from Papau based on its Woodley E (1991). (Ed.) Medicinal Plants of Papua New Guinea. Part I: physical characteristics and chemical composition. Biodiversitas. Morobe Province. Wau Ecology Institute Handbook No.11. Verlag Josef 13:124-129. Margraf, Germany. 158 pp. Newman DJ, Gragg GM (2007). Natural products as sources of new Zure D, Maiguo E, Wau JS (2010). Piper bolanicum Chew from Goung drugs over the last 25 years. J. Nat. Prod. 70(3): 461-477. Mountain Range, Kabwum District, Papua New Guinea: Preliminary Perez C, Pauli M, Bazerque P (1990). An antibiotic assay by agar well Report on a Psychoactive Plant. Environment Papua New Guinea. diffusion method. Acta Biologiae et Medecine Experrimentaalis. 15: 2(2):15-18. 113-115. Prakash R (2009). Phytochemical observation and antibacterial activity of Cyperus escuentus. Anc. Sci. Life. 28(4):16-20. Cite this article as:

Rates MK (2001). Plants as sources of drugs. Toxicon. 39(5): 603-613. Wagi R, Gusamo BK (2019). Antimicrobial activity of crude oil Rohman A, Sugeng R, Che Man YB (2012). Characterization of red fruit extracts from fruits of Cynometra ramiflora Linn and Pandanus (Pandanus conoideus Lamk) oil. Int. Food Res. J. 19(2): 563-567. conoideus Lamk from Papua New Guinea against ten pathogenic Rohman A, Windarsih A (2018). Characterization, biological activities, micro-organisms. Acad. J. Med. Plants. 7(10): 236-242. and authentication of red fruit (Pandanus conoideus Lam) oil. Food Res. 2(2):134-138. Submit your manuscript at: Sakulas HW, Harakuwe AH, Rungwa S, et al (2010). The ‘Chill-Thaw’ http://www.academiapublishing.org/ajmp Method for Noni Juice Extraction. Environment Papua New Guinea. 2(2): 31-33.