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In Vitro Bioactive Potential of an Ethnomedicinal Mangrove Plant, (Heritiera Fomes Buch

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In Vitro Bioactive Potential of an Ethnomedicinal Mangrove Plant, (Heritiera Fomes Buch Indian Journal of Geo-Marine Sciences Vol. 44(5), May 2015, pp. 704-713 In Vitro bioactive potential of an ethnomedicinal mangrove plant, (Heritiera fomes Buch. Ham.) from Odisha Coast, India Jayanta Kumar Patra1,2 & Hrudayanath Thatoi1* 1Department of Biotechnology, College of Engineering and Technology, Biju Patnaik University of Technology, Bhubaneswar-751003, Odisha, India 2School of Biotechnology, Yeungnam University, Gyeongsan-712-749, Republic of Korea *[E-mail: [email protected]] Received 05 February 2014; revised 05 March 2014 Heritiera fomes, a mangrove species occurring in mangrove forest along Odisha coast, India is known for its ethnomedicinal uses for the treatment of gastrointestinal, hepatic and skin diseases etc. All the four solvent (acetone, ethanol, methanol and aqueous) extracts of leaf and stem powder showed antibacterial activity (11 mm to 22 mm) at 50 mg/ml concentration. Leaf extracts possess better antioxidant properties (90% scavenging) as compared to stem extracts with dose dependency. Phytochemical screening of extracts showed the presence of phenols, cardiac glycosides, terpenoids etc. Overall results indicated that extracts of H. fomes might be applicable in natural medicine and pharmaceutical industries for drug discovery. [Keywords: Bioactivity, Heritiera fomes, mangrove plants, phytochemicals] Introduction “Sundari” is one of the most extensively occurring Mangroves are salt tolerant forest ecosystems true mangrove species along Odisha coast, India and found mainly in the tropical and subtropical the leaves, stem and bark have been traditionally used intertidal regions of the world largely confined to the for the treatment of various diseases such as region between 30º North and South of the equator1. gastrointestinal disorders (including dysentery, They are good source of herbal medicine because they diarrhoea, indigestion, colic, acidity, constipation, can grow under stressful conditions such as water bloating, lack of appetite, stomach ache), hepatic logging, fluctuating tidal and salinity etc. where no disorders (including jaundice, hepatitis); skin diseases vascular plants can survive and thus they produce (including eczema, abscess, acne, boils, scabies, several secondary metabolites which are of medicinal itch, infections, dermatitis, rash, sores, scar, importance2. Many mangrove plants are used in warts), diabetes and goiter9,10. Although H. fomes has traditional medicine being important source of shown some ethnomedicinal properties, there are no phytochemicals, natural antioxidants and studies focusing on its antioxidant and antimicrobial antimicrobials agents as reported by several authors3-6. activities of solvent extracts. Consequently, the Such bioactivity of the mangrove plants can be objective of the present study is to determine the assigned to phytochemicals present in the extracts7. antimicrobial and antioxidant activities of H. fomes. It Since mangrove plants possess bioactive is also aimed to screen the phytochemicals with a antimicrobial substances, which are safer to use view to explore their possible application in without any side effects, there is need to screen out pharmaceutics. more mangrove plants with potent antimicrobial activities. Apart from being a source of Materials and Methods antimicrobial compounds, mangroves are also Fresh, young and tender leaves and stem of potential sources of natural antioxidants and some of Heritiera fomes Buch.-Ham. (Sterculiaceae) were them are used as food8. collected from the mangrove forests along Odisha The Heritiera fomes commonly known as coast, India (Figure 1). The specimens were identified PATRA AND THATOI: IN VITRO BIOACTIVE POTENTIAL OF MANGROVE PLANT, HERITIERA FOMES 705 at Department of Natural Products, Institute of the method of Oyaizu17. The DPPH (2,2-diphenyl -1- Minerals and Materials Technology, Bhubaneswar picryl hydrazyl) radical scavenging effect was (RRL-B), Odisha, India and voucher specimen (VS determined by following modified methods of Hatano No. RRL-B-12568) was deposited. The extracts of the et al.18. The ferrous ion chelating activity was dried powder of the leaves and stem of the plant were assessed as described by Zhao et al.19. Nitric Oxide prepared using solvents (acetone, ethanol, methanol scavenging activity was assessed by Griess reaction and water). Liquid extracts were evaporated to method20. dryness by vacuum distillation and stored at 4°C for A qualitative phytochemical test to detect the further analysis. presence of alkaloid, tannin, saponin, flavonoid, cardiac glycosides, sterols, anthroquinone glycosides, carbohydrates and protein was carried out using standard procedures21 and quantitative phytochemical test for determination of proteins, carbohydrates, flavonoid, alkaloids, riboflavin, thiamine, and tannins21 and nitrogen, potassium, phosphorous, potassium pentoxide, potassium dioxide was carried out using standard procedures22. Statistical Analysis Experiments were carried out in triplicates and the data was expressed as mean value ± standard deviation. Where applicable, the means of all the parameters were examined for significance by two Fig. 1— Map showing study sites way analysis of variance (ANOVA) and the differences between samples were determined by Duncan’s Multiple Range test using GenStat In vitro antimicrobial activity of leaf and stem discovery (edition 3) statistical software package. extracts of H. fomes were carried out against nine Differences were considered significant at a human pathogenic bacteria and three fungi strains. probability level of P<0.05. Correlation analysis Bacteria used in the study were obtained from between different antioxidant assays and between Institute of Microbial Technology, Chandigarh and TAC, PC and ASA were carried out using the others including the fungi were lab isolates. Agar cup correlation programme in MINITAB Software. The plate method of Khalid et al.11 was carried out to regression analysis between the DPPH radical establish the antibacterial activity of all the four scavenging and Nitric oxide scavenging assay were solvent extracts against the test pathogens. The also carried out using MINITAB Software. antifungal activity study was carried out by agar well diffusion method of Perez et al.12. Minimal inhibitory concentration (MIC) was determined by two fold Results microdilution method13. Two antibiotics viz. The results of the antimicrobial screening of the Neomycin (30µg/disc) and Norfloxacin (10µg/disc) solvent extracts of leaf and stem of H. fomes against for bacteria and antifungal agent, clotrimazole (100 ten pathogenic bacteria and three fungal strains are μg/ml) for fungus were used as standard. summarized in Table 1 and Figure 2 (antimicrobial); Total phenolic content (PA) was estimated Table 2 (MIC and MBC) and Table 3 (antifungal) according to the methods of Slinkard and Singleton14 respectively. Different extracts of leaf and stem of H. using catechol as standard phenolic compound. fomes was found to be active against all the Ascorbic acid content (ASA) was estimated following pathogenic bacteria tested except the aqueous stem the methods of Barros et al.15 with slight extract which was inactive against Bacillus brevis. modifications. Total antioxidant capacity (TAC) of Zone of inhibition of the H. fomes extract against plant extracts was determined by Prieto et al.16. The different pathogenic bacteria varied between 11 mm reducing power of solvent extracts was determined by to 22 mm (Table 1 and Figure 2). 706 INDIAN J. MAR. SCI., VOL. 44, NO. 5 MAY 2015 Table 1: Antibacterial activity of solvent extracts of leaf and stem of Heritiera fomes and standard antibiotics Leaf Stem Antibiotic Name Neomycin Norfloxacin Strains Acetone Ethanol Methanol Aqueous Acetone Ethanol Methanol Aqueous (30µg/disc) (10µg/disc) S. aureus 21a±1.4 18cd±0.49 18cd±0.21 16de±0.35 19no±1.4 20n±0.71 19no±0.21 18o±0.49 17± 0.70 33± 0.35 S. flexneri 19b±0.71 17d±0.49 14ef±0.21 15e±0.21 18o±0.21 19no±0.21 17op±0.49 15pq±0.21 18± 0.35 30± 1.4 B. licheniformis 18b±0.35 17d±0.49 15e±0.71 12f±0.49 15pq±0.49 17op±0.71 16p±0.35 17op±0.71 21± 1.4 30± 0.35 B. brevis 20ab±0.71 16de±0.21 17d±0.49 15e±0.71 14q±0.35 13q±0.49 13q±0.21 0±0 27± 0.35 33± 0.70 V. cholerae 21a±0.49 17d±0.71 15e±0.35 14ef±0.35 13q±0.49 14q±0.21 13q±0.35 14q±0.49 18± 0.70 36± 0.70 P. aeruginosa 21a±0.21 19c±0.71 17d±0.49 13f±0.49 18o±0.49 17op±0.71 16p±0.49 18o±0.49 18± 0.35 40± 0.35 S. epidermidis 19b±0.49 17d±0.49 16de±0.71 12f±0.49 16p±0.71 17o±0.21 15pq±0.71 22m±1.4 15± 0.07 35± 0.70 B. subtilis 15d±0.49 16de±0.49 14ef±0.21 13f±0.35 17op±0.49 16p±0.71 16p±0.49 18o±0.49 16± 1.4 36± 0.07 E. coli 18b±0.35 18cd±0.21 16d±0.49 13f±0.35 16p±0.21 18o±0.21 16p±0.21 16p±0.35 14± 0.07 35± 0.35 useful antimicrobial compounds as the MIC values ranged in between 1.25-5.0 mg/ml (Table 2). The results of the antifungal activity of H. fomes screened against three pathogenic fungi along with a standard antifungal agent (Clotrimazole, 100 µg/ml) are given in Table 3. Both the methanol and aqueous leaf extracts of H. fomes were active against the fungal strains (7 mm to 14 mm) (Table 3). However, the activity of the plant extract is comparatively less than that of the standard antifungal agent. Methanol extract of leaf of H. fomes showed significantly higher antifungal activity against C. kruzii with zone of inhibition of 14 mm, in comparison to other extracts and fungal strains at p<0.05 (LSD-0.506). Several antioxidant assays were performed on different solvent extracts of H. fomes to study the Fig. 2— Antibacterial activity of extracts of H. fomes. antioxidant potential of the plant and its parts.
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