GJRMI - Volume 5, Issue 1, january 2016 by Dr. Hari Venkatesh.K.Rajaraman

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INDEX – GJRMI - Volume 5, Issue 1, January 2016 MEDICINAL PLANTS RESEARCH Bio-diversity ECOLOGICAL STATUS AND CONSERVATION OF RARE PLANTS IN HIGH ALTITUDE LANDSCAPES OF INDIAN WESTERN HIMALAYA K N Singh, Gopichand*, Brij Lal, N P Todaria

01–18

Natural & Life sciences ANTIOXIDANT AND ANTIMICROBIAL ACTIVITIES OF NUMIDICA GROWING IN BABOR MOUNTAINS FROM ALGERIA

ENDEMICTREE ABIES

Ghadbane Mouloud*, Bounar Rabah, Khellaf Rebbas, Medjekal Samir, Belhadj Hani, Benderradji Laid, Smaili Tahar, Harzallah Daoud 19–28

Ethno-Medicine THE USE OF VARIOUS PLANT TYPES AS MEDICINES BY LOCAL COMMUNITY IN THE ENCLAVE OF THE LORE-LINDU NATIONAL PARK OF CENTRAL SULAWESI, INDONESIA Rosmaniar Gailea*, Ach. Ariffien Bratawinata, Ramadhanil Pitopang, IrawanWijaya Kusuma

29–40

INDIGENOUS MEDICINE Ayurveda - Panchakarma - Case Study SIRAVYADHEM IN VARICOSE VEINS – A CASE STUDY Shona Rudolph*, Prashanth D, Ananthram Sharma

41–48

COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – INFLORESCENCE OF SANTALUM ALBUM L.* OF THE FAMILY SANTALACEAE PLACE – OFF KANAKAPURA ROAD, BANGALORE, KARNATAKA, INDIA *BOTANICAL NAME VALIDATED FROM www.theplantlist.org AS ON 10/01/2016

Research article ECOLOGICAL STATUS AND CONSERVATION OF RARE PLANTS IN HIGH ALTITUDE LANDSCAPES OF INDIAN WESTERN HIMALAYA K N Singh1, Gopichand2*, Brij Lal3, N P Todaria4 1,2,3

Biodiversity Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh, 176 061, India 4 Forestry Department, HNB Garhwal University, Srinagar, Uttarakhand, India *Corresponding author: E-mail: gopichand@ihbt.res.in

Received: 06/10/2015; Revised: 25/12/2015; Accepted: 05/01/2016

ABSTRACT The study has been carried out in Lahaul-Spiti region (13,835 km2) of western Himalaya, India. Various landscape element (LSE) types dominated by higher plants in the entire Lahaul-Spiti were identified and subjected for stratified random sampling using quadrat method to study the species distribution patterns, status of threatened and rare plants and their assessment for their for long-term monitoring and conservation. A total 703 quadrats were laid in Lahaul and 841 quadrats in Spiti. Within the LSEs studied in Lahaul, a total 288 species of higher plants belonging to 174 genera distributed among 51 families were recorded. Whereas, in Spiti, a total 166 plant species, represented by101 genera and 37 families were recorded. Species richness and plant families exhibited a significant and negative correlation with altitude in both Lahaul and Spiti. Taxa showing the highest population density in the entire Lahaul-Spiti were Aconitum heterophyllum, Angelica glauca, Aquilegia fragrans, Corydalis govaniana, Delphinium brunonianum, Heracleum lanatum, Hyssopus officinalis, Jurinea macrocephala and Saussurea glanduligera. During the study, a total of 18 and 9 threatened species were found from Lahaul and Spiti respectively. Similarly endemic taxa observed in the sampling units in Lahaul were 85 which were higher than that of Spiti where 52 endemic species were found. A total 58 plants in Lahaul and 57 plants in Spiti were identified as rare which need immediate conservation for their sustainability. Considering the ecological status and rarity of various plant species, present study suggests the priority for the conservation of their natural populations distributed in the higher elevations of Lahaul-Spiti, Indian western Himalaya. Key words: Alpine landscape, Conservation, Ecological monitoring, Indian Himalaya, Rare plants

Cite this article: K N Singh, Gopichand, Brij Lal, N P Todaria (2016), ECOLOGICAL STATUS AND CONSERVATION OF RARE PLANTS IN HIGH ALTITUDE LANDSCAPES OF INDIAN WESTERN HIMALAYA, Global J Res. Med. Plants & Indigen. Med., Volume 5(1): 01â&#x20AC;&#x201C;18

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 1 | January 2016 | 01–18

INTRODUCTION Indian Himalaya, bio-geographically a unique zone in the world, has the maximum degree of endemism in the Asian region and is also considered a storehouse for most of the valuable rare medicinal and aromatic plants (Trivedi, 2002). A total 1,748 species of medicinal plants have been reported from IHR (Samant et al., 1998) of which 25.3% are endemic, which are represented in varying life forms at various altitudes and habitats. Unfortunately, many of these species are critically endangered today, threatened by the both anthropogenic impacts and climate change (Singh, 2014). Various reports show around 90% of raw material of medicinal plants for herbal industry in India and for export is drawn from natural habitats (Gupta et al., 1998; Ved et al., 1998). Studies on population status and ecological attributes of plant species play a vital role to formulate the conservation plans (Singh 2008; Singh et al., 2008a; Uniyal et al., 2002) and data generated through such studies are of high significance that are required to develop appropriate strategies for their long term monitoring and sustainable use. Therefore, approaches for prioritizing strategies for conservation and monitoring of plants should be based on quantitative information of their population structure, habitat characteristics and intensity of anthropogenic pressure. Distribution pattern in various habitats and numerical estimates of even the local wild populations of these plants are not available, as no such detailed investigations have been done so far in the region (Singh 2008; Kala 2000). Even today, available literature doesn‟t throw any light on the ecological status and factors governing their distribution at different scales even in well-known groups of taxa, as it has hardly been attempted in many parts of the Indian Himalayan range. Himachal Pradesh situated between 30°22/ to 33°12/ north latitude and 75°47/ to 79°04/ east longitude, is an important area of prime

biodiversity concern in the Indian western Himalaya. Due to varied climate and ecological landforms ranging from 350–6975 m elevation above mean sea level, the State harbours rich plant diversity. Besides sharing about 23.52% of total flowering plant species in the country, Himachal Pradesh is considered a veritable emporium of medicinal, aromatic and endemic taxa (Chauhan, 2006). In the high altitude alpine and cold desert zones of the State, Lahaul-Spiti harbours a wide range of rare, endemic, threatened and medicinally important species of higher plants (Singh and Brijlal 2008c; Singh et al., 2012d). Hence, it can be considered an important hot spot in western Himalayan Region. Lahaul-Spiti has its own ecological significance due to its diversified landscapes with an average elevation of 4000 m above msl. Nearly 985 plant species belonging to 353 genera and 79 families of angiosperms and gymnosperms have been reported from the area (Aswal and Mehrotra 1994), which represents more than 82% of the total threatened plant species of Himachal Pradesh according to the IUCN (2015) status categories (Anonymous 2003). Over extraction of valuable plants in Lahaul-Spiti has also been experienced resulting depletion of their population in nature (Kala 2000). Some of the rare and endangered medicinal plantshave been reported from Spiti valley (Singh et al., 2008a, 2008b) but the ecological studies on quantitative attributes of vegetation in various landscape elements (LSEs) for the entire region have not been reported so far. Keeping above points in consideration, present investigation has focused on the ecological studies of rare and threatened higher plant communities distributed in different landscape elements of Lahaul-Spiti region. Keeping in view the importance of ecological studies, the present investigation was aimed at with the objective of selecting certain plant species based on their ecological and cultural significance for long-term monitoring and conservation for sustainable utilization.

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Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 1 | January 2016 | 01–18

MATERIAL AND METHODS Study area The current study was carried out in Lahaul-Spiti region of Indian western Himalaya, India (Fig. 1). Lahaul-Spiti, situated between 31 44'57''–32 59'57''N latitudes and 76 46'29''–78 41'34''E longitudes. The area is

an ideal landscape, encompassing most of the diverse ecosystems widely spread between temperate to cold desert zones. With a total area of 13,835 sq km, Lahaul and Spiti remain isolated from each other by Kunjum Pass (a mountain peak; 4591 m) for more than six months in a year due to heavy snowfall.

Figure 1. Map of study area (Lahaul-Spiti) showing major localities

Lahaul valley is enclosed by main Himalayan ranges on the North, Pir Panjal range on the South, the Kunjum range which separates Lahaul from Spiti on the East, and the off-shoot of the Pir Panjal range on the West and Rohtang Pass (3955 m) that connects Lahaul to Kullu district of Himachal Pradesh. Spiti valley is situated beyond the Kunjum Pass (4591 m) and lies in adjoining part of the central Himalaya in the rain shadow of the Great Himalayan ranges and is bounded by Tibet in the North-East, Kinnaur and Kullu districts of Himachal Pradesh in the South-East and West, and Ladakh (Jammu and Kashmir) in the North. High mountains which enclose the district act as a barrier against monsoon rains. Therefore, there are occasional showers during

the monsoon period but the regular snowfall usually starts from the middle of November and continues till the beginning of April. Generally, July and August are the hottest months where the highest day temperature goes upto 30.5 C in Spiti valley and 33.2 C in Lahaul. January and February are the coldest months where the lowest temperature reaches upto −30.5 C in Spiti and −14.81 C in Lahaul. Vegetation mainly comprises of alpine/ cold desert types in Lahaul-Spiti except subalpine and temperate vegetation in Lahaul in the Northwestern part of the district. The temperate and sub-alpine vegetation is mainly dominated by coniferous evergreen forests and broad-leaved deciduous species, however, alpine vegetation comprises of herbaceous species and few scattered shrubs (Singh, 2008).

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 1 | January 2016 | 01–18

Field sampling Ecological surveys were conducted periodically between June and November of each year from 2004 to 2007. During this period, vegetation comes in full bloom as the snow melts due to the increase in day temperature. Community structure of higher plants in each LSE was assessed by stratified random sampling as suggested by Misra (1968), and Mueller-Dombois and Ellenberg (1974) employing quadrat method. A minimal sample area was determined using species area curve for sampling purpose so that species composition is adequately represented during random quadrat sampling (Misra 1968; Mueller-Dombois and Ellenberg 1974). The adequacy of sample size was estimated by stopping sampling at the point at which additional quadrats didn‟t significantly affect the mean of species. The size of quadrat was related to the size and spacing of the individuals in the sampling plot. The random quadrats of 10m × 10m for tress species, 5m × 5m for shrubs and 1m × 1m for herbaceous species were laid during sampling of plants. The scanty and sparse vegetations on steep slopes and barren terrain were sampled following altitudinal interval of 300 m. During the ecological surveys, a total of 703 quadrats covering all the LSEs were laid across 2550 to 4950 m in Lahaul whereas a total 841 quadrats were laid at elevations from 3100 to 5200 m in Spiti valley. Among various plants including endemic taxa, i.e, those which are unique to habitas of western Himalayan region, a number of species have been selected for their rarity status, taking into consideration the following criteria: wide distribution range of species with narrow habitat specificity; low population structure; locally abundant over a large range in specific habitat; constantly sparse in a specific habitat; abundance in specific habitats but in localized pockets with restricted distribution to special

habitats (Nayar 1996; Tandon 1998; Anonymous 2003; Singh 2008). In addition, species having a distribution range only within 300 m altitudinal interval along with threatened status as per IUCN (2015-3) recommendations were also considered for rarity. Species richness and population density Data on presence and absence of plant species in each LSE were analysed for estimation of frequency, however numerical strength of species in each LSE was computed in terms of density as suggested by Mishra (1968). Species richness was expressed in terms of total number of taxa present in sampling plots in each LSE. Therefore, species richness is the total number of species in each LSE. Collection specimens

and

identification

plant

From each sampling site, the voucher specimens of plant species observed in the quadrats, were collected and identified with the help of local and regional floras (Polunin and Stainton 1984; Aswal and Mehrotra 1994). Herbarium of Botanical Survey of India (BSD), Dehradun was also consulted for the identification and authentication of the plant species. Specimens collected during the surveys were processed in the herbarium laboratory following Jain and Rao (1976). These were pressed, dried in blotting sheets and poisoning with formaldehyde or mercuric chloride solution (0.5%) was done to protect against insect and fungal attack before mounting on the herbarium sheets. The plant specimens having unique voucher number were deposited in the herbarium of the Institute of Himalayan Bioresource Technology (PLP), Palampur, Himachal Pradesh and one set of these specimens has also been deposited in the herbarium of BSI, Dehradun for future reference.

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Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 1 | January 2016 | 01–18

Statistical tests To test the significance of variation in various ecological parameters at 5 per cent level of significance (p<0.05), the data were subjected to the statistical analysis using CPCS-1 (Windows version). To test the null hypothesis of lack of significant correlation, absolute values of correlation co-efficients (r) were compared with the critical values given at various degrees of freedom at 0.05 and 0.01 levels of significance. Degrees of freedom considered were n-2, where n is the number of sample. If the absolute value of „r‟ exceeded this critical value, then correlation was considered significant and the null hypothesis was rejected (Ludwig and Reynold, 1988). RESULTS Species richness and distribution pattern During current study, a total 166 species belonging to 101 genera and 37 families of higher plants were recorded in Spiti, where the highest species numbers were found in the following plant families: Asteraceae (30), Fabaceae (18), Polygonaceae (11), Rosaceae and Lamiaceae (9 each), Scrophulariaceae (8),

Ranunculaceae (7), Gentianaceae and Apiaceae (6 each), Poaceae, Chenopodiaceae, Brassicaceae and Boraginaceae (5 each). Each of the remaining 24 families had fewer than 5 species. However, results revealed the distribution of 288 species (174 genera and 51 families) in Lahaul valley. Like Spiti, maximum number of species in Lahaul was also contributed by Asteraceae (57) followed by Polygonaceae and Rosaceae (18 each), Poaceae (17), Lamiaceae (15), Apiaceae, Fabaceae and Scrophulariaceae (14 each), and Ranunculaceae (13). However, overall species richness in most of the families at Lahaul was higher than the families found in Spiti. In Spiti valley, species richness as well as number of genera and families increased significantly from the lowest altitudinal interval of 3100–3400 m to the one above (3400–3700 m) and then gradually decreased with elevation up to the uppermost altitudinal band (4900– 5200 m) (Fig. 2-A). Along the elevation gradient, distribution of species richness (r=0.775; p<0.05) and genera (r=-0.803; p<0.05) was observed significantly negative. Plant families also revealed similar trend, however level of significance was comparatively low (r=-0.713; p<0.1).

Figure 2. Distribution pattern along an altitudinal gradient; (A) Spiti and (B) Lahaul

160

200 180

140 r=-0.775 (p<0.05)

160

r=-0.751 (p<0.05)

120 Species richness Genera Plant family

Number

120 100 r=-0.803 (p<0.05)

Number

140

100 80

r=-0.713 (p<0.1) 20

Species richness Genera Plant family

r=-0.807 (p<0.05)

r=-0.854 (p<0.01)

0 31003400

34003700

37004000

40004300

43004600

46004900

49005200

25502850

28513150

31513450

Altitudinal gradient (m)

(A)

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

34513750

37514050

40514350

Altitudinal gradient (m)

(B)

43514650

46514950

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 1 | January 2016 | 01–18 Figure 3. Distribution trend of endemic and threatened plants along altitude; (A) Spiti and (B) Lahaul

50 45 40

Endemic plant s

T hreatened plant s

20 15 10

Number of species

Num ber of species

Threatened plants

35 30

Endemic plants

25 20 15 10 5

5 0

31003400

3401- 37013700 4000

40014300

4301- 46014600 4900

49015200

2550- 2851- 3151- 3451- 3751- 4051- 4351- 46512850 3150 3450 3750 4050 4350 4650 4950

Altitudinal gradient (m)

(A)

(B)

Figure 4. Number of species under different categories of threats along altitude; (A) Spiti and (B) Lahaul

4.5

7 6

Endangered Critically Endangered

2.5

Vulnerable 2 1.5

Number of species

3.5

Endangered Critically Endangered Vulnerable

5 4 3 2

0.5 0

0 31003400

34013700

37014000

40014300

43014600

46014900

49015200

Altitudinal gradient (m)

2550- 2851- 3151- 3451- 3751- 4051- 4351- 46512850 3150 3450 3750 4050 4350 4650 4950 Altitudinal gradient (m)

(A)

Maximum species richness, number of genera and families in Lahaul were recorded at the intermediate elevation (3150–3450 m). Like Spiti, correlation along altitudinal gradient in Lahaul was also observed in similar pattern, where species richness (r=-0.751; p<0.05), genera (r=-0.807; p<0.05) and plant families (r=-0.854; p<0.01) exhibited a significantly negative relationship (Fig. 2-B). Another point showing the peakness of species richness and genera was observed at 3750–4050 m due to their significant increase from the previous altitudinal band of 3450–3750 m. At higher elevations, a number of medicinal and aromatic plant species growing in alpine

(B)

regions such as Aconitum rotundifolium, Allium auriculatum, Artemisia macrocephala, Biebersteinia odora, Corydalis govaniana, Draba lasiophylla, Dracocephalum heterophyllum, Gentianella tenella, Leontopodium brachyactis, Nepeta longibracteata, Pedicularis rhinanthoides, Rhodiola wallichiana, Saxifraga flagellaris, Saussurea bracteata, S. gnaphalodes, S. nana, Waldheimia glabra and many others were observed with limited distribution in localized pockets. Dominant genera which contributed maximum species at extreme elevations were Saussurea, Rhodiola, Nepeta, Polygonum and Potentilla. The maximum number of plant

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 1 | January 2016 | 01–18

species at the highest elevation in the study area was represented by the families Asteraceae, Crassulaceae, Lamiaceae, Poaceae, Polygonaceae, Ranunculaceae and Rosaceae.

Threatened plants were not found in sampling plots between 4301–4600 m and above 4900 m. Arnebia euchroma, an important critically endangered medicinal plant in Spiti was found from 3401–4300 m on dry slopes. Dactylorhiza hatagirea, an endangered orchid was observed from 3401–4000 m in marshy areas and along river banks. Among endangered plants, Ephedra gerardiana was widely distributed from 3100–4900 m on dry slopes. Hyoscyamus niger was found only near human habitations between 3400 and 3700 m, however, Bergenia stracheyi was observed on the moist rocky slopes in the elevation band of 3700–4300 m.

Status of threatened and rare plants Total number of plants of threatened category observed in different LSEs was found higher in Lahaul (18) as compared to Spiti (9) (Fig. 5 & 6: Table 1). The highest proportion of threatened species and endemic plants in Spiti was found in the middle elevation range of 3700–4000 m and above this, the proportion declined with increasing altitude (Fig. 3-A).

Table 1. Rare and threatened plant species in Lahaul-Spiti, western Himalaya Family/Taxa

Life forms

Endemism*

Distribution extent along elevation (m)

Subdivision

IUCN** status [2015-3]

Apiaceae Angelica glauca Edgew.

Herb

Endemic

300

Endangered

Carum carvi Linn.

Herb

387

Chaerophyllum acuminatum Lindl.

Herb

Endemic

Localized

Ferula jaeschkeana Vatke

Herb

Endemic

Localized

L,S

Vulnerable

Heracleum lanatum Michx.

Herb

Endemic

267

Vulnerable

Pleurospermum stellatum C.B. Clarke

Herb

Localized

P. stylosum Benth. ex Clarke

Herb

Localized

L,S

Selinum vaginatum C.B. Clarke

Herb

Endemic

Localized

Asteraceae Artemisia japonica Thumb.

Herb

Endemic

A. maritima Linn.

Herb

Endemic

928

A. parviflora Roxb.

Herb

Localized

A. scoparia Waldst. & Kit.

Herb

A. tournefortiana Reichb.

Herb

Localized

A. molliusculus (Lindl. ex DC.) Clarke

Herb

Cremanthodium plantagineum (Franch.) Aswal Erigeron alpinus Linn.

Herb

Localized

Herb

Endemic

Localized

E. borealis (Vierch.) Simmons

Herb

Localized

E. multiradiatus (DC.) Benth.

Herb

Localized

Hieracium crocatum Fries.

Herb

Endemic

H. umbellatum Linn.

Herb

Localized

Leontopodium brachyactis Gand.

Herb

Localized

Jurinea macrocephala (Decne.) Benth.

Herb

Endangered

Leontopodium fimbrilligerum J.R. Drum

Herb

Localized

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 1 | January 2016 | 01â&#x20AC;&#x201C;18 L. himalayanum DC.

Herb

192

Saussurea bracteata Decne.

Herb

Endemic

Localized

S. glanduligera Sch.- Bip. ex Hook.f.

Herb

Endemic

Localized

S. gnaphalodes (Royle) Sch.-Bip.

Herb

Localized

S. nana (Pamp.) Pamp.

Herb

Endemic

Saussurea costus (Falc.) Lipschitz

Herb

Endemic

Localized

S. gnaphalodes (Royle) Sch.-Bip.

Herb

Localized

Tanacetum falconeri Hook.f.

Herb

Localized

L,S

Tragopogon pratense Linn.

Herb

Localized

Waldheimia glabra (Decne.) Regel

Herb

Endemic

W. tomentosa (Decne.) Regel.

Herb

Endemic

Localized

Biebersteiniaceae Biebersteinia odora Staph. ex Fisch.

Herb

Berberidaceae

Berberis pseudumbellata Parker

Shrub

Endemic

262

B. lycium Royle

Shrub

Endemic

Localized

Least Concern

Betulaceae Betula utilis D. Don

Tree

Boraginaceae

Arnebia euchroma (Royle) T.M. Johnston

Herb

775

L, S

A. guttata Bunge

Herb

170

Critically Endangered -

Brassicaceae Draba lasiophylla Royle

Herb

Endemic

Localized

D. setosa Royle

Herb

Localized

Lepidium latifolium Linn.

Herb

166

Capparidaceae Capparis spinosa Linn.

Herb

Endemic Caprifoliaceae

Lonicera asperifolia (Decne.) Hook.f. & Th. L. spinosa Jacq. ex Walp.

Shrub

Localized

Shrub

Endemic

Localized

Viburnum cotinifolium D. Don

Shrub

Localized

Crassulaceae Rhodiola himalensis (D. Don) Fu

Herb

Localized

R. wallichiana (Hook.) Fu

Herb

Endemic

Sedum tibeticum Hook.f. & Th.

Herb

Localized

Elaeagnaceae Hippophae rhamnoides Linn.

Shrub

373

L,S

Vulnerable

H. tibetana Schlecht.

Shrub

Localized

L,S

Ephedraceae Ephedra gerardiana Wall. ex Stapf.

Herb

Endemic

1700

L,S

Endangered

E. regeliana Florin.

Herb

170

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Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 1 | January 2016 | 01â&#x20AC;&#x201C;18 Ericaceae Cassiope festigiata (Wall.) D. Don

Shrub

Endemic

Localized

Rhododendron anthopogon D. Don

Shrub

Endemic

Localized

Vulnerable

R. campanulatum D. Don

Shrub

Endemic

Localized

Vulnerable

Fabaceae Astragalus himalayanus Klotz.

Herb

Endemic

171

A. strobiliferus Royle

Herb

Localized

Thermopsis inflata Camb.

Herb

Endemic

337

Fumariaceae Corydalis cornuta Royle

Herb

Localized

Corydalis govaniana Wall.

Herb

Endemic

Localized

L,S

C. meifolia wall.

Herb

Localized

Gentianaceae Gentiana detonsa (Rottb.) Ma

Herb

Localized

Gentianella moorcroftiana (Wall. ex G. Don) Airy-Shaw Gentianopsis paludosa (Hook.f.) Ma

Herb

Endemic

300

L,S

Herb

Localized

Jaeschkea canaliculata (Royle ex G. Don) Knobl. J. oligosperma (Griseb.) Knolb.

Herb

Endemic

Localized

Herb

Swertia ciliata (G. Don) Burtt.

Herb

322

L,S

Geraniaceae Geranium wallichianum D. Don ex Sweet

Herb

Endemic Lamiaceae

Dracocephalum heterophyllum Benth.

Herb

656

Hyssopus officinalis Linn.

Herb

119

Nepeta floccosa Benth.

Herb

Endemic

Localized

Nepeta longibracteata Benth.

Herb

Endemic

Localized

L,S

Liliaceae Allium carolinianum Wall.

Herb

Localized

Allium jacquemontii Kunth

Herb

Localized

Trillidium govanianum (D. Don) Kunth

Herb

Localized

179

267

L,S

Endangered

300

Endangered

Oleaceae Fraxinus xanthoxyloides Wall. ex DC.

Shrub

Endemic Orchidaceae

Dactylorhiza hatagirea (D. Don) Soo

Herb

Podophyllum hexandrum Royle

Herb

Endemic Podophyllaceae Polygonaceae

Rheum emodi Wall. ex Meissn.

Herb

Endemic

L,S

Endangered

R. spiciforme Royle

Herb

Endemic

1409

L,S

Vulnerable

Ranunculaceae Aconitum heterophyllum Wall. ex Royle

Herb

Endemic

506

Endangered

A. rotundifolium Kar. & Kir.

Herb

Localized

L,S

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Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 1 | January 2016 | 01–18 Aquilegia fragrans Benth.

Herb

Endemic

276

Delphinium brunonianum Royle

Herb

Endemic

Localized

374

L,S

Vulnerable

Saxifragaceae Bergenia stracheyi (Hook.f. & Th.) Engl.

Herb

Endemic Scrophulariaceae

Pedicularis pectinata Wall. ex Benth.

Herb

Endemic

267

L,S

P. punctata DC.

Herb

Endemic

Localized

P. rhinanthoides Schrenk. ex Fisch. & Mey

Herb

Localized

Picrorhiza kurrooa Royle ex Benth.

Herb

Endemic

767

Endangered

Localized

L,S

Endangered

Localized

Solanaceae Hyoscyamus niger Linn.

Herb

Valerianaceae

Valeriana himalayana Grub.

Herb

Endemic Violaceae

Viola canescens Wall. ex Roxb.

Herb

Localized: distribution in < 5 m range; L-Lahaul; S-Spiti *Endemism: Nayar (1996) and Kala (2005); **IUCN Status: 2015-3

Distribution of Rheum emodi and Ferula jaeschkeana was confined between 3700– 4000 m and 3400–4000 m respectively. Among vulnerable plants, occurrence of Hippophae rhamnoides and Rheum speciforme was found between 3400–3700 m and 3700–4000 m elevation. In Lahaul, number of threatened and endemic plants was observed maximum in between 3150–3450 m elevation (Fig.3-B) in similar trend as shown in the distribution of species richness, which further decreased with increasing elevation. At 3750–4050 m, another maxima in the proportion of endemic taxa was also observed which further decreased towards the extreme elevations. In the lowest elevation band (2550–2850 m), threatened plants were not recorded. Among critically endangered species, distribution of Aconitum heterophyllum was observed in between 3150–3750 m, whereas Arnebia euchroma was found in elevation range of 4050–4350 m. Plants belonging to endangered category distributed in between 2850–3450 m were Angelica glauca and Podophyllum hexandrum on moist slopes, however, distribution of Dactylorhiza hatagirea was confined in marshy areas only.

Distribution of Picrorhiza kurrooa was observed at 3150–3450 m elevation and also from 3750–4350 m; however it was restricted in small patches on moist slopes. Hyoscyamus niger was confined at 3150–3450 m and Rheum emodi in between 3750–4050 m elevation range. Jurinea macrocephala was found at 3750–4050 m and 4650–4950 m altitudinal bands. At the highest altitudinal interval (4650– 4950 m), among endangered species, distribution of Ephedra gerardiana was observed on dry slopes. Distribution of plants belonging to vulnerable category was observed from 2850–4650 m. Occurrence of Heracleum lanatum was recorded from 2850 to 3450 m, Bergenia stracheyi from 3150–3750 m and Ferula jaeschkeana at 3450–3750 m. Individuals of Rheum spiciforme were found in dry areas having boulders at 2850–3150 m and from 4050–4650 m. Shrubs belonging to vulnerable category were Hippophae rhamnoides, Rhododendron anthopogon and R. campanulatum, where distribution of H. rhamnoides was observed between 2850– 3450 m along the river sides, Rhododendron campanulatum at 3150–3450 and R. anthopogon at 3750–4050 m.

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Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 1 | January 2016 | 01â&#x20AC;&#x201C;18 Figure 5. Some of the threatened taxa found in Lahaul-Spiti

A. Podophyllum hexandrum

B. Ephedra gerardiana

C. Hippophae rhamnoides

D. Arnebia euchroma

E. Aconitum heterophyllum

F. Dactylorhiza hatagirea

G. Hippophae tibetana

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Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 1 | January 2016 | 01â&#x20AC;&#x201C;18 Figure 6. Some of the rare plants in Lahaul-Spiti

A. Saussurea bracteata

B. Saussurea gnaphalodes

C. Ephedra regeliana

E. Waldheimia glabra

D. Saussurea nana

F. Allium auriculatum

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G. Hyssopus officinalis

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Results revealed that most of the threatened plants recorded in both Lahaul and Spiti valleys were from the endangered category followed by the plants of vulnerable class (Fig. 4-A and 4-B). Distribution of endemic plants was observed at all the altitudinal intervals in Lahaul and Spiti. Total number of endemic plants recorded in Lahaul was 85 and was higher than that of Spiti where a total 52 endemic taxa were found distributed. Besides these, a total 58 plants in Lahaul and 57 plants in Spiti have been evaluated as rare, which also include various endemic and threatened taxa. Most of these plants are medicinal and aromatic in nature and were observed in small habitats with concentrated patches in localized pockets in both Lahaul and Spiti (Table 1). Some of the common taxa identified as rare belonging to both Lahaul and Spiti valleys were Aconitum rotundifolium, Artemisia japonica, A. maritima, Bergenia stracheyi, Corydalis govaniana, Dactylorhiza hatagirea, Ephedra gerardiana, Ferula jaeschkeana, Gentianella moorcroftiana, Geranium wallichianum, Hippophae rhamnoides, H. tibetana, Hyoscyamus niger, Nepeta longibracteata, Pedicularis pectinata, Pleurospermum stylosum, Rheum emodi, R. spiciforme, Saussurea gnaphalodes and Tanacetum falconeri. The unique rare taxa found only in Lahaul were Aconitum heterophyllum, Allium jacquemontii, Angelica glauca, Artemisia parviflora, Berberis pseudumbellata, B. lyceum, Betula utilis, Carum carvi, Cassiope festigiata, Chaerophyllum acuminatum, Corydalis meifolia, Cremanthodium plantagineum, Delphinium brunonianum, Fraxinus xanthoxyloides, Gentiana detonsa, Gentianopsis paludosa, Heracleum lanatum, Jaeschkea canaliculata, J. oligosperma, Jurinea macrocephala, Leontopodium fimbrilligerum, L. himalayanum, Lonicera spinosa, Picrorhiza kurrooa, Pleurospermum stellatum, Podophyllum hexandrum, Rhododendron anthopogon, R. campanulatum, Saussurea costus, Selinum vaginatum, Swertia ciliata, Thuja orientalis, Tragopogon pratense,

Trillidium govanianum, Valeriana himalayana, Viburnum cotinifolium and Viola canescens. In Spiti, the rare plants identified were Arnebia guttata, Artemisia molliusculus, A. scoparia, A. tournefortiana, Astragalus himalayanus, A. strobiliferus, Biebersteinia odora, Capparis spinosa, Corydalis cornuta, Draba lasiophylla, D. setosa, Dracocephalum heterophyllum, Ephedra regeliana, Erigeron alpinus, E. borealis, E. multiradiatus, Hieracium crocatum, H. umbellatum, Hyssopus officinalis, Leontopodium brachyactis, Lepidium latifolium, Lonicera asperifolia, Nepeta floccosa, Pedicularis punctata, P. rhinanthoides, Rhodiola himalensis, R. wallichiana, Saussurea bracteata, S. glanduligera, S. nana, Sedum tibeticum, Thermopsis inflata, Waldheimia glabra and W. tomentosa. Taxa showing the highest population density of less than 2 individuals/m2 in the entire Lahaul-Spiti were Aconitum heterophyllum, Angelica glauca, Aquilegia fragrans, Corydalis govaniana, Delphinium brunonianum, Heracleum lanatum, Hyssopus officinalis, Jurinea macrocephala and Saussurea glanduligera. DISCUSSION Present investigation through light on species richness and their distribution in various landscape elements in alpine cold desert of Lahaul-Spiti region in Indian western Himalaya. In comparison with earlier reports (Singh et al., 2008a; Singh et al., 2008b; Singh 2012e), it was also observed that some of the important species showed low and dwindling population in their natural habitats in LahaulSpiti . Distribution pattern of these species was observed generally localized in concentrated patches because most of these were restricted to marshy and moist areas, dry scrub, rocks, boulder, undulating terrain and alpine meadows and thus suggest their habitat specificity. Observations on concentrated patches, habitat preferences, low population structure, anthropogenic activities including various other ecological features of most of the medicinal plants have been supported by the findings of

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Ghimire et al., (2005) and Bhatt et al., (2006) in Himalayan region. Drastic change in vegetation due to ecological variables is a fundamental characteristic of mountain ecosystems. These are steep climate gradients where species richness changes over relatively short distances due to change in various specific climatic parameters and by limitation of resources (Theurillat et al., 2003). Diversity of microhabitats supporting variety of species ranging from temperate to dry cold alpine zone with varying degrees of slopes, altitudes and many other ecological variables besides anthropogenic activities may be responsible for the significant variation in species richness. At higher elevations, the life form diversity is reduced drastically, perhaps due to harsh arid environment (Pauses and Austin, 2001). Therefore, significant negative correlation of species richness, genera and plant families in relation to altitude observed in Lahaul and Spiti explains that species richness pattern in this region largely depends upon altitude which is one of the basic gradients along which lifeform changes (Singh, 2008a). The maximum species richness, number of threatened species and endemic plants at intermediate altitudes in Lahaul (3150–3450 m) and Spiti (3400–3700 m), decreasing above is in line with the review of Rahbek (1995) who found that half of the studies showed maximum species richness at intermediate elevations. Other than 3150–3450 m interval, another increase in species richness and genera at 3750–4050 m in Lahaul could be due to the transitional communities where occurrence of species of both sub-alpine and pure cold arid zone is possible in the sampling plots. Bhattarai and Vetaas (2006) and Kharkwal et al. (2005) also found similar patterns of plant species richness along the Himalayan elevation gradient, with maximum species richness at mid-altitudes where overlapping of neighboring communities occurs and immigration of species from adjacent species pool could become possible in most common habitats.

The low frequency of occurrence of threatened medicinal plants reveals their localized or narrow distribution restricted to certain habitats in the study area. It could be the reason that threatened taxa were not available in sampling plots at the lowest elevation interval (2550–2850 m) in Lahaul and from 4300 m to 4600 m and above 4900 m in Spiti. The over-extraction of medicinal plants from natural habitats as raw material for herbal industries (Gupta et al., 1998; Ved et al., 1998) is another reason for their low availability (Uniyal et al., 2006). In high altitude cold desert landscapes of Lahaul-Spiti, Singh (2008) also reported continuous over-extraction resulting in the depletion of populations of many rare endemic and endangered plants. Kala (2000, 2005) and Singh et al. (2008c, b) also reported over harvesting of plants for sale in local market and collection to prepare indigenous medicines, an important factor to play a role in depletion of their number in wild habitats. Among plant parts, leaves have been reported to be used to a large extent as a remedy followed by roots and flowers to cure various ailments in the region. Among threatened plants, Ephedra gerardiana, Rheum spiciforme and Arnebia euchroma were widespread at different elevations, but the distribution was restricted to fragmented habitats, which is one of the reasons for their rarity. Due to overexploitation and habitat degradation, the original habitats are fragmented into isolated patches, which also lead to the fragmentation of populations of plant species (Tandon, 1998). In such fragmented systems, smaller fragments will initially contain more species. However, the rate of extinction of species in such smaller groups, with specific habitat requirements, is faster than for the species with a continuous broad habitat range, because ecological niches available for survival are reduced (Tandon 1998; Nautiyal et al., 2002). Keeping in mind this mechanism, a number of endemic, rare and threatened plants observed in Lahaul-Spiti need immediate attention for conservation of their natural habitats and localized populations. Taxa

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showing the highest population density of less than 2 individuals/m2 in the entire Lahaul-Spiti (Aconitum heterophyllum, Angelica glauca, Aquilegia fragrans, Corydalis govaniana, Delphinium brunonianum, Heracleum lanatum, Hyssopus officinalis, Jurinea macrocephala and Saussurea glanduligera) suggest the priority conservation measures of natural populations of these species. Long-term monitoring and conservation implications Various developmental activities are causing a great loss to the biodiversity in the Indian Himalayan region, where medicinal plants in particular are declining at faster rate due to their over exploitation for trade. It is believed that excessive anthropogenic activities are the main cause of decline in the population and availability of medicinal plants in the Himalayan region (Dhar et al., 2000). All these activities in Lahaul-Spiti might have led to the depletion of a number of valuable plant species. Considering the ecological features, rarity, population status and medicinal importance of most of the plant species, present study recommends the creation of Conservation Areas in the entire Lahaul-Spiti to conserve certain localities having maximum species richness for sustainability. Sites under high anthropogenic activities, having low species diversity also needed to be covered under this plan. It was observed that continuous pressure of tourism, trampling, grazing, and accumulation of waste material is an alarming risk to population of plant bioresource at various localities like Rohtang Pass, Kaza, Baralacha etc. These activities are responsible for habitat degradation and may become the cause of species extinction from their niche areas. Initiating the concept of ecotourism, demarcation of grazing routes to avoid trampling and habitat degradation, awareness of the tourists, local people and local doctors regarding the valuable plant resources and danger of their unsustainable availability due to over exploitation, besides strengthening the

ecological studies may become some of the very effective efforts for conservation. It is suggested to prepare micro-plans for each commercially important species suggesting harvesting methods and quantity to be harvested which are inadequately known for most of medicinal plants. Multiplication of the plants using tissue culture techniques and conventional methods for their transplantation in natural habitats and niche areas of the species will be a significant initiative in the direction of their conservation. Development of agro-techniques of certain species for LahaulSpiti to meet the requirement of raw material for commercial use will help to reduce the pressure on the existing populations in natural habitats. Besides including all the plants identified as rare in Lahaul-Spiti in the conservation plans, a few potential plant species have been proposed for long-term monitoring and conservation which include Aconitum heterophyllum, Aconitum rotundifolium, Allium carolinium, Angelica glauca, Arnebia euchroma, Arnebia guttata, Artemisia maritima, Capparis spinosa, Carum carvi, Dactylorhiza hatagirea, Dracocephalum heterophyllum, Ephedra gerardiana, Ephedra regeliana, Ferula jaeschkeana, Hippophae rhamnoides, Hippophae tibetana, Hyssopus officinalis, Jurinea macrocephala, Picrorhiza kurrooa, Podophyllum hexandrum, Rheum emodi, Rheum spiciforme, Saussurea costus and Inula racemosa. Cultivation of these valuable species needs to be promoted in the line of Saussurea costus and Inula racemosa which are being cultivated in many of the localities of Lahaul for ages. The only way out is conservation in natural habitats and domestication to meet the growing demand of medicinal plants. In this connection cultivation of Aconitum heterophyllum, Carum carvi and Dactylorhiza hatagirea by the local people in Trilokinath and Teenghrat areas of Udaipur (Lahaul) is an inspiring effort to conserve the valuable germplasm for sustainability.

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ACKNOWLEDGEMENTS Author is thankful to Director, CSIRInstitute of Himalayan Bioresource Technology, Palampur, for the facilities and Dr. R.D. Singh, for critical suggestions. National Bioresource Development Board (Dept. of

Biotechnology), Government of India, is duly acknowledged for financial support. First author is also thankful to the Council of Scientific and Industrial Research, New Delhi, for the award of Senior Research Fellowship.

REFERENCES Anonymous (2003). Threat assessment and management prioritization of selected medicinal plants of West Himalayan States. Foundation for Revitalization of Local Health Traditions (FRLHT), Bangalore, India. Aswal, BS, Mehrotra, BN (1994). Flora of Lahaul-Spiti. Bishen Singh Mahendra Pal Singh, Dehradun, India. Bhatt, A, Rawal, RS, Dhar, U (2006). Ecological features of a critically rare medicinal plant, Swertia chirayita, in Himalaya. Plant Species Biology 21:49–52. Bhattarai, KR, Vetaas, OR (2006). Can Rapoport‟s rule explain tree species richness along the Himalayan elevation gradient, Nepal? Diversity and Distributions 12: 373–378. Chauhan, NS (2006). Medicinal and Aromatic Plants of Himachal Pradesh. Indus Pub, New Delhi, India. Dhar, U, Rawal, RS, Upreti, J (2000). Setting priorities for conservation of medicinal plants-a case study in the Indian Himalaya. Biological Conservation 95: 57–65. Ghimire, SK, Doyle, M, Thomas, A (2005). Conservation of Himalayan medicinal plants: harvesting patterns and ecology of two threatened species, Nardostachys grandiflora DC. and Neopicrorhiza scrophulariiflora (Pennel) Hong. Biological Conservation 124: 463–475.

Gupta, A, Vats, SK, Brij Lal (1998). How cheap can a medicinal plant species be? Current Science 74: 555–556. Jain, SK, Rao, RR (1976). A handbook of field and herbarium methods. Today and Tomorrow Printers and Publishers, New Delhi, India. Kala, CP (2000). Status and conservation of rare and endangered medicinal plants in the Indian trans- Himalaya. Biological Conservation 93: 371–379. Kala, CP (2005). Indigenous uses, Population density, and conservation of threatened medicinal plants in protected areas of the Indian Himalaya. Conservation Biology 19(2): 368–378. Kharkwal, G, Mehrotra, P, Rawat, YS, Pangtey, YPS (2005). Phytodiversity and growth in relation to altitudinal gradient in the central Himalayan (Kumaun) region of India. Current Science 89(5): 873–878. Ludwig, JA, Reynolds, JF (1988). Statistical ecology. A primer on methods and computing. John Wiley and Sons, New York. Misra, R (1968). Ecological workbook. Oxford and IBH Publishing Company, Calcutta, India. Mueller-Dombois, D, Ellenberg, H (1974). Aims and methods of vegetation ecology. John Wiley and Sons, New York.

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Nautiyal, BP, Prakash, V, Bahuguna, R, Maithani, U, Bisht, H, Nautiyal, MC (2002). Population study for monitoring the status of rarity of three aconite species in Garhwal Himalaya. Tropical Ecology 43(2): 297–303. Nayar, MP (1996). “HOT SPOTS” of endemic plants of India, Nepal and Bhutan. Tropical Botanic Garden and Research Institute (Palode), Thiruvananthapuram, India. Pauses, JG, Austin, MK (2001). Patterns of plant species richness in relation to different environments: an appraisal. Journal of Vegetation Science 12: 153– 166. Polunin, O, Stainton, A (1984). Flowers of the Himalaya. Oxford University Press, Delhi, India. Rahbek, C (1995). The elevation gradient of species richness: a uniform pattern? Ecography 18: 200–205. Samant, SS, Dhar, U, Rawal, RS (1998). Biodiversity status of a protected area of west Himalaya. 1-Askot Wildlife Sanctuary. International Journal of Sustainable Development and World Ecology 5:194–203. Singh, KN (2008). Ecological studies on landscape elements, species diversity and ethnobotany of higher plants in Lahaul-Spiti region of western Himalaya. Ph.D thesis submitted to HNB Garhwal University Srinagar, Garhwal, India. Singh, KN (2012e). Spatial distribution and ecological status of higher plants in Indian western Himalaya. Paper presented in 65th British Columbia Institute of Agrologists Conference held in Victoria, BC, Canada.

Singh,

RB (2014). Indigenous uses of medicinal and edible plants of Nanda devi biosphere reserve – a review based on previous studies. Global J Res. Med. Plants & Indigen. Med. 3(2): 57–66.

Singh, KN, Brij Lal (2008c). Ethno-medicines used by tribal communities against four common ailments in Lahaul-Spiti, western Himalaya. Journal of Ethnopharmacology 115: 147–159. Singh, KN, Brij Lal, Todaria, NP (2008b). Observations on dwindling population of Arnebia euchroma, a critical endangered taxon in Himachal Pradesh. Journal of Non Timber Forest Products 15 (3): 157–160. Singh, KN, Brij Lal, Todaria, NP (2012d). Ethnobotany of higher plants in Spiti cold desert of western Himalaya. Nature and Science 10(5). Singh, KN, Gopichand, Kumar, A, Brij Lal, Todaria, NP (2008a). Species diversity and population status of threatened plants in different landscape elements of the Rohtang pass, western Himalaya. Journal of Mountain Science 5: 73–83. Tandon, V (1998). Need for CAMP for LahaulSpiti and Ladakh, the conservation of medicinal plants. In: Ved, D.K. and Tandon, V. (eds.). Conservation assessment and management plan workshop for high altitude medicinal plants of north west Himalayas. Foundation for Revitalization of Local Health Traditions, Bangalore, India. Theurillat, JP, Schlussel, A, Geissler, P, Guisan, A, Velluti, C, Wiget, L (2003). Vascular plants and bryophyte diversity along elevation gradients in the Alps. In: Nagy L, Grabherr G, Korner CH, Thompson DBA (eds.). Alpine biodiversity in Europe. pp. 185–193, Springer, Berlin.

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Trivedi, PC (2002). Ethnobotany: An overview. Aavishkar, Publishers, Distributers, Jaipur, India Uniyal, SK, Awasthi, A, Rawat, GS (2002). Current status and distribution of commercially exploited medicinal and aromatic plants in upper Gori valley, Kumaon Himalaya, Uttaranchal. Current Science 82(10): 1246–1252.

Source of Support: National Bioresource Development Board (Dept. of Biotechnology), Government of India

Uniyal, SK, Kumar, A, Brij Lal and Singh, RD (2006). Quantitative assessment and traditional uses of high value medicinal plants in Chhota Bhangal area of Himachal Pradesh, western Himalaya. Current Science 91(9): 1238–1242. Ved, DK, Tandon, V (eds.) (1998). CAMP report for high altitude medicinal plants of Jammu-Kashmir and Himachal Pradesh. Foundation for Revitalization of Local Health Traditions, Bangalore, India.

Conflict of Interest: None Declared

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Research article ANTIOXIDANT AND ANTIMICROBIAL ACTIVITIES OF ENDEMICTREE ABIES NUMIDICA GROWING IN BABOR MOUNTAINS FROM ALGERIA Ghadbane Mouloud1*, Bounar Rabah2, Khellaf Rebbas3, Medjekal Samir4, Belhadj Hani5, Benderradji Laid6, Smaili Tahar7, Harzallah Daoud8 1,2,3, 4,6,7

Department of Natural and Life Sciences, Faculty of Sciences, University of Mohamed Boudiaf-M’sila, PO Box 166 Ichebilia, M’sila, 28000, Algeria. 1,5,8 Laboratory of Applied Microbiology, Department of Microbiology, Faculty of Natural and Life Sciences, University Ferhat Abbas, Setif 1 19000, Algeria. * Corresponding Author: Email: mouloud_ghadbane@yahoo.fr

Received: 10/12/2015; Revised: 05/01/2016; Accepted: 06/01/2016

ABSTRACT The aim of present study was to determine the antimicrobial and antioxidant activity of essential oil and different extracts from Abies numidica leaves. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay was used to detect in vitro antioxidant activity.Antibacterial activity was tested against nine bacterial species, representing both Gram positive and Gram negative bacteria. Antifungal activity was evaluated using two Candida species. The results indicate that the crude methanolic extracts and the essential oil resulted in the highest antioxidant activities compared to other solvents.All extracts of A. numidica showed antimicrobial activity against both Gram positive and Gram negative bacteria, as well as fungi, especially methanolic extracts and essential oil. A. numidica could be exploited for the isolation of bioactive compounds which could be a potential source for antioxidant and antimicrobial activities. KEY WORDS: Antifungal activity, DPPH, essential oil, bioactive, methanolic extracts.

Cite this article: Ghadbane Mouloud, Bounar Rabah, Khellaf Rebbas, Medjekal Samir, Belhadj Hani, Benderradji Laid, Smaili Tahar, Harzallah Daoud (2016), ANTIOXIDANT AND ANTIMICROBIAL ACTIVITIES OF ENDEMICTREE ABIES NUMIDICA GROWING IN BABOR MOUNTAINS FROM ALGERIA, Global J Res. Med. Plants & Indigen. Med., Volume 5(1): 19–28

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INTRODUCTION The beneficial effect of antioxidants and antibiotics on the maintenance of health in human has become an important subject that has engaged many scientists across the world over the last decade. Antioxidants have significant inhibition roles, not only on undesirable changes in the flavor and nutritional quality of food, but also on tissue damage in various human diseases such as inflammation, cancer, and atherosclerosis (Rajendran et al., 2014). To avoid or delay this autoxidation process, conventional artificial antioxidants such as butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT), propyl gallate (PG) and tertiary butyl hydroquinone (TBHQ) have been used for more than five decades. However, these synthetic antioxidants have been suspected to cause serious negative health effects. Recently synthetic products are being restricted in the industries, because of harmful effects observed such as human toxicity and environmental pollution. For this reason, there is a growing interest in studies of natural additives as potential antioxidants (Roby et al., 2013). The substitution of synthetic antioxidants by plant materials has caused great interest in nutrition research. The development of bacterial resistance to presently available antibiotics necessitated the search for new antimicrobial agents. The antimicrobial activity of plant oils and extracts has formed the basis of many applications, including raw and possessed potential as natural agents for food preservation, pharmaceuticals, alternative medicine and natural therapies (Dung et al., 2008) During the last few decades, the global interest in the study of various medicinal plants has increased rapidly due to their antibacterial and antioxidant activities, low toxicity and the potential to be a cheaper alternative to costly synthetic drugs (Benderradji et al., 2014; Farjana et al., 2014).

The family Pinaceae, one of the major conifer taxa, is divided into several genera of which Abies, Picea and Pinus are among the largest. Conifers of the genus Abies Mill. play an important ecological role in forest ecosystems which cover large parts of the northern hemisphere. Currently 51 species have been described in the genus Abies; of these, ten species and subspecies are distributed around the Mediterranean Sea (Liepelt et al., 2010). The fir of Numidia (Abies numidica De Lannoy ex Carrière) is an endemic plant growing in Algeria, and is often used as folk medicines. A. numidica appears only on the tops of mountains of Babor and Tababort from Algeria, where these Forests cover only a few hundred hectares (Quezel, 1985, 1998). However, to the best of our knowledge, there are so far no published reports on bioactivity of this plant. The aim of this work was to evaluate the antioxidant and antimicrobial activities of the different extracts of Abies numidica. MATERIALS AND METHODS Plant material: Abies numidica is a tree growing up to 20 m tall. Whorled branches, spread horizontally, the last erect. The leaves are needle-like, moderately flattened, glossy dark green with a patch of greenish-white stomata near the tip above, and with two greenish-white bands of stomata below. The cones are glaucous green with a pink or violet tinge, maturing brown, 13–20 cm long (Quezel and Santa, 1962; Schutt and Lang, 1991). Aerial parts of A. numidica were collected during Marsh 2015 from mountains of Babor in Algeria (Figure 1). Plant materials were identiﬁed by Dr. Bounar Rabah according to “Now Flora of Algeria” (Quezel and Santa, 1962) (Figure 2) and voucher specimens deposited in the Department of Natural and Life Sciences, Faculty of Sciences, Mohamed Boudiaf University, M’sila, Algeria. Preparation of extracts: Leaves of A. numidica were air-dried at room temperature and then ground at approximately 0.2 to 0.4 mm. The dried powder (100 g) was extracted

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by soaking with 1 L of 70% (v/v) methanol for 24 h at room temperature. After filtration, the residue was processed similarly with the same amount of solvent. The crude methanol extract (MeOH extract) was concentrated to dryness under reduced pressure at 40°C with a rotary evaporator, and stored at 4°C until further use. Five grams from the methanol extract was

further fractionated by successive solvent extraction with ethyl acetate (EtOAc fraction), and then with n-butanol saturated water (nBuOH fraction).The crude extract using purified water was subjected to successive appropriate solvent with increased polarity (ethyl acetate and n-butanol) under a continuous reﬂux setup in a Soxhlet extractor.

Figure 1: A map showing the site from which the Abies numidica leaves were collected.

Figure 2: Fir of Numidia (Abies numidica).

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Essential oil: Air-dried plant materials (leaves) were subjected to hydrodistillation using a Clevenger type apparatus (Clevenger, 1928), according to the method recommended in the European Pharmacopoeia 4 (C E, 2002). The essential oil was dried over anhydrous sodium sulphate and then stored in sealed glass vials at 4°C prior to analysis. The yield of essential oil was determined in triplicate. DPPH free radical scavenging assay: The evaluation of the free radical-scavenging activity of essential oil, crude ethanol extract, ethyl acetate fraction and n-butanol fraction was based on the measurement of the reducing ability of antioxidants toward the 1,1-diphenyl2-picrylhydrazyl (DPPH) 2,2-diphenyl-1picrylhydrazyl DPPH radical.Scavenging of DPPH radical was assayed following the method of Dung et al. (2008), with some modification. Solutions with different extract concentrations were prepared and 25 µL of each solution was added to 150 µL of DPPH solution (63.4µM) and 125 µL of 96 % ethanol. The mixture was left to stand for 1h in the dark at room temperature. Reduction of the amount of the DPPH radical present was measured using a decrease in the absorption value at 517 nm. The extract concentration providing 50% inhibition (IC50) was calculated using a graph of the scavenging effect percentage against the extract concentration. The scavenging effect percentage was calculated from the formula:

(Bacillus subtilis ATCC 10876, Klebsiella pneumoniae ATCC532, Staphylococcus aureus ATCC 6538, S. epidermidis ATCC 12228, Enterococcus faecalis ATCC 29212, Pseudomonas aeruginosa ATCC27853, Pseudomonas syringaepv. Tomato 1086, Escherichia coli ATCC25922, Micrococcus luteus ATCC 533) and two strains of fungi (Candida albicans ATCC 24433 and Candida tropicalis R2 CIP 203). For experiments, the bacterial strains were grown on Müeller-Hinton agar at 37°C for 18 hours whereas Sabouraud dextrose agar at 35°C for 24 hours was used for yeasts.

Determination of antimicrobial activity

Disc diffusion assay: Antimicrobial activity of A. numidica essential oil, crude methanol and solvent extracts was evaluated by the disc diffusion test, according to the standard M2-A8 from Clinical Laboratory Standards Institute ( CLSI, 2003) for bacteria and according to NCCLS (2004), document M44-A for yeasts. Briefly, the essential oil and the different test solvent extracts (methanol, ethyl acetate and nbutanol) were loaded to the sterilized sterile 6 mm discs, and then the impregnated discs with 50 μl at a concentration of 100 mg/ml was prepared with dimethyl sulphoxide (DMSO). DMSO was used as control. The inoculum was prepared in a sterile solution of 0.85% sodium chloride and the optical density of the suspension was adjusted to 0.5 McFarland (1 × 108 colony forming units (CFU)/mL). Then the Müeller-Hinton Agar plates for bacteria and Mueller-Hinton Agar + 2% Glucose and 0.5 µg/mL Methylene Blue Dye (GMB) medium for yeasts were inoculated, allowed to dry and the discs previously prepared were placed over the agar. The plates were incubated at 37°C for 48 h under microaerobic conditions for bacteria and at 35°C for 20 to 24 hours of incubation for yeasts and after incubation the diameter of the inhibition zone was measured. All the assays were done in triplicates and the results were given in mean ± SD. Standard antibiotics such as gentamicin for pathogenic bacteria and fluconazole for pathogenic fungi served as positive controls.

Test microorganisms: Antimicrobial activity was tested against nine strains of bacteria

Statistical analysis: All experiments were repeated 3 times. The data are expressed as the

, where ADPPH is the absorbance of a negative control (blank sample containing the same amount of solvent and DPPH solution) and AS is the absorbance of the sample. IC50 values denote the concentration of sample, which is required to scavenge 50% of DPPH free radicals. The IC50 values were calculated from linear regression analysis. Butylated hydroxyl toluene (BHT) was used as a positive control.

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mean ± SD (standard deviation). Analysis of variance (ANOVA) was carried out to determine significant differences by the SAS statistical software version 9.0 (SAS Institute, Inc., Cary, NC, USA). The significance of the difference was checked by the Duncan test, and differences were considered significant at p < 0.05.

RESULTS AND DISCUSSION Total yield: The yields of the different solvent extracts and the essential oil obtained are presented in Table1. The results demonstrated a signiﬁcant (P < 0.05) difference in the extract yields among the solvent systems used.

Table 1: Extraction yield and antioxidant activity of the crude methanolic extract, fractions and essential oil of A. numidica leaves. Samples Crude methanolic extract n-butanol fraction ethylacetate fraction Essential oil BHT

Yield (%) 13.123 ± 0.554a 3.823 ± 0.305b 2.503 ± 0.411c 0.260 ± 0.036d -

Scavenging activity on DPPH IC50 (mg/mL) 0.282 ± 0.004c 0.424 ± 0.018b 0.554 ± 0.025a 0.288 ± 0.024c 0.143 ± 0.031d

*: Values in the table are means of three independent experiments and error bars indicates standard deviation of the mean. Letters show significant deference using Duncan’s test (p <0.05).

From this table (1), we noted that the extraction yields with methanol resulted in the highest amount of total extractable compounds whereas the lowest yields were obtained in essential oil. In fact, this variation in the yields can be attributed to the polarities of the different compounds present in the leaves of A. numidica. The extraction yield of the essential oil was 0.260 ± 0.036 % in the dry basis, which was less than that reported before (Ramdani et al., 2014). Differences in the extraction yields could be derived from several factors such as genetic factors, environmental conditions, sample pretreatment, extraction methods and extraction conditions (Cho et al., 2011; Khoudja et al., 2014; Dong et al., 2015). DPPH free radical scavenging activity: The antioxidant potential of essential oil, methanol, ethyl acetate, and n-butanol extracts of A. numidica was evaluated on the basis of their ability to scavenge stable free DPPH radicals. The concentration required to attain 50% radical-scavenging effect (IC50) was determined from the results of a series of concentrations tested. A lower IC50 value corresponds to a larger scavenging activity. Scavenging activity increased with the increase in the concentration of essential oil

and different extracts of A. numidica leaves. Crude methanolic extract and essential oil showed signiﬁcantly higher activity than the other extracts, as the amount of sample required to decrease the DPPH concentration by 50 % (IC50) was found to be 0.282 and 0.288 mg/mL for crude methanolic extract and essential oil, respectively (Table 1). The experimental data reveals that these extracts and essential oil are likely to have the effect of scavenging free radical. However, the IC50 value for extracts and essential oil were much lower than BHT used in the present study. The difference in preparation of the extracts might affect DPPH radical activity of them. Phenolic compounds such as flavonoids are known to be potential antioxidants due to their ability to scavenge free radicals and active oxygen species such as singlet oxygen, superoxide anion radical and hydroxyl radicals (Orak et al., 2011; Djouossi et al., 2015). Therefore, the presence of such compounds could be responsible for the antioxidant activity found in the crude methanolic extract and the essential oil. To our best knowledge this is the first report on characterization of antioxidant properties of A. numidica.

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Table 2: Antimicrobial activity of the essential oil and different extracts of the leaves of Abies numidica. Experimental strains

B. subtilis ATCC 10876 S. aureus ATCC 6538 E. faecalis ATCC 29212 S. epidermidis ATCC 12228 M. luteus ATCC 533

Essential oil

Mean zone of inhibition (mm)* Crude Ethyl acetate n-butanol methanolic fraction fraction extract Gram-positive bacteria

Gentamicin

14 ± 1.73b

28.33 ± 1.53a

10.33 ± 0.58c

0.0 ± 0.0d

13.33 ± 1.53b

13.33 ± 0.58d

47.67 ± 2.52a

24.67 ± 2.08b

0.0 ± 0.0e

19.17 ± 0.58c

41.67 ± 2.89a

44.33 ± 1.15a

25.33 ± 1.53b

12.33 ± 1.53d

17.33 ± 1.15ce

25.00 ± 1.0a

10.0 ± 1.0d

0.0 ± 0.0e

13.33 ± 1.55c

18.67 ± 1.15b

25.0 ± 1.0b

52.67 ± 2.52a

28.67 ± 4.16b

10.67 ± 1.55c

24.33 ± 0.58b

Gram-negative bacteria E. coli ATCC 25922 P. aeruginosa ATCC 27853 P. syringae pv. tomato 1086 K. pneumoniae ATCC 532

8.67 ± 1.15c

14.00 ± 1.0a

0.0 ± 0.0d

11.0 ± 1.0b

12.33 ± 1.53b

18.67 ± 1.53a

13.33 ± 1.53b

0.0 ± 0.0d

10.33 ± 1.53c

24.33 ± 1.53b

29.67 ± 1.53a

25.67 ± 2.08b

14.67 ± 0.58c

13.0 ± 1.73c

12.87 ± 0.42b

12.67 ± 1.15hb

0.0 ± 0.0d

9.33 ± 0.58c

21 ± 1.0a

Yeasts C. albicans ATCC 24433 C. tropicalis R2 CIP 203

Fluconazole

43.33 ± 2.31a

45.33 ± 1.53a

23.33 ± 1.15c

0.0 ± 0.0d

35.33 ± 0.58b

30.0 ± 1.73b

33.67 ± 2.52a

19.67 ± 1.53c

0.0 ± 0.0d

28.33 ± 1.53b

*: Values in the table are means of three independent experiments and error bars indicates standard deviation of the mean; a, b, c letters show significant deference using Duncan’s test (p <0.05).

Antimicrobial activity: The results from the antimicrobial activity of the essential oil and the three extracts tested by disc diffusion method are summarized in Table 2. The essential oil and all extracts of A. numidica

showed an inhibitory effect on the growth of all tested microbial strains. The rate of inhibition was determined by measuring the diameter of the clear zone around the soaked ﬁlter paper (Table 2 and Figure 3).

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Figure 3: Antibacterial activities of essential oil and extracts of Abies numidica based on inhibition zone (mm).

The diameters of inhibition zones ranged from 52.67 ± 2.52 to 8.67 ± 1.15 mm. The best result was obtained for methanolic extract and the essential oil (Table 2). The variation in antimicrobial activity between different tested bacterial and fungal strains using essential oil extracts, fluconazole and gentamicin (P < 0.05) was statistically significant. Essential oil and different extracts of A. numidica showed significant antibacterial activity, especially against M. luteus, S. aureus and E. faecalis, a recognized pathogenic species. The oil and different extracts also efficiently inhibited the growth of C. albicans and C. tropicalis.

Antimicrobial activity could be due to the presence of a higher concentration of the alcohol-soluble active molecules in A. numidica. The effectiveness of essential oil and extracts against a wide range of microorganisms is related to their hydrophobicity, which enables them to integrate into the lipids of the cell membrane and mitochondria, rendering them permeable and leading to leakage of cell contents (Horvathova et al., 2014). The antimicrobial results of current study are in accordance with literature, where extracts of many Abies species were effective against pathogenic bacteria and

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fungi (Bağci and Diğrak, 1996; Vishnoi et al., 2007; Xia et al., 2012; Ramdani et al., 2014; Poaty et al., 2015). Previous studies clearly revealed that the essential oil and different extracts have strong activity against bacteria and yeast, because these phenols and flavonoids have a strong antimicrobial potential (Hossain et al., 2014; Al-Jadidi and Hossain, 2015; Stefanović et al., 2015). Therefore, it is necessary to conduct a more profound research on the relationship between antimicrobial activity and chemical structure of each phenol and flavonoid compound in the A. numidica extract. CONCLUSION The present study has shown that the crude methanol extract and essential oil of A. numidica gave the highest antioxidant activity, followed by the n-butanol fraction and the ethyl acetate fraction was the least active. The crude methanol extract and essential oil, proven to have very good potential as a source of antimicrobial activity.

In summary, the current study proved in vitro antioxidant and antimicrobial activity of A. numidica essential oil and different extracts. The identification of the bioactive compounds and study of mechanisms of actions are necessary prior to application. Further research should be done in order to determine the relation-ship between bioactivities and chemical structure of each compound in the A. numidica extracts. ACKNOWLEDGEMENTS: This work was funded by the Algerian Ministry of Higher Education and Scientific Research. We greatly acknowledge Department of Natural and Life Sciences, Faculty of Sciences, University of Mohamed BoudiafM’sila and Laboratory of Applied Microbiology, Department of Microbiology, Faculty of Natural and Life Sciences, University Ferhat Abbas-Setif, Algeria for their constant encouragement.

REFERENCES Al-Jadidi, H. S. K., & Hossain, M. A. (2015). Studies on total phenolics, total flavonoids and antimicrobial activity from the leaves crude extracts of neem traditionally used for the treatment of cough and nausea. Beni-Suef University Journal of Basic and Applied Sciences, 4(2): 93–98. Bağci, E. & Diğrak, M. (1996). Antimicrobial Activity of Essential Oils of some Abies (Fir) Species from Turkey. Flavour and Fragrance Journal, 11(4): 251–256. Benderradji, L., Rebbas, K., Ghadbane, M., Bounar, R., Brini, F., &Bouzerzour, H. (2014). Ethnobotanical study of medicinal plants in Djebel messaad region (M'sila, Algeria).Global J Res. Med. Plants & Indigen. Med. 3(12): 445–459.

Cho, M., Lee, H., Kang, I., Won, M., & You, S. (2011). Antioxidant properties of extract and fractions from Enteromorpha prolifera, a type of green seaweed. Food Chem., 127: 999–1006. Clevenger, J. F. (1928). Apparatus for the determination of volatile oil. J. Am. Pharm. Assoc., 17: 341–346. CLSI (2003). CLSI (Clinical and Laboratory Standards Institute) CLSI document M2-A8-Standardization of the antimicrobial susceptibility tests for disc-diffusion; approved standard (8th ed.), Vol. 23, Wayne, PA, USA. C E (2002). Council of Europe, European Pharmacopoeia Commission European Directorate for the Quality of Medicines & Healthcare: European Pharmacopoeia. 4th edition. Strasbourg.

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Djouossi, M. G., Tamokou, J.-d.-D., Ngnokam, D., Kuiate, J.-R., Tapondjou, L. A., Harakat, D. &VoutquenneNazabadioko, L. (2015). Antimicrobial and antioxidant flavonoids from the leaves of Oncoba spinosa Forssk. (Salicaceae). BMC Complementary and Alternative Medicine, 15,134: 1–8. Dong, H., Zhang, Q., Li, L., Liu, J., Shen, L., Li, H., & Qin, W. (2015). Antioxidant activity and chemical compositions of essential oil and ethanol extract of Chuanminshen violaceum. Industrial Crops and Products, 76: 290–297. Dung, N. T., Kim, J. M., & Kang, S. C. (2008). Chemical composition, antimicrobial and antioxidant activities of the essential oil and the ethanol extract of Cleistocalyx operculatus (Roxb.) Merr and Perrybuds. Food and Chemical Toxicology 46: 3632–3639. Farjana, A., Zerin, N., &Kabir, M.d. S. (2014). Antimicrobial activity of medicinal plant leaf extracts against pathogenic Bacteria. Asian Pac J Trop Dis, 4(2): 920–923. Horvathova, E., Navarova, J., Galova, E., Sevcovicova, A., Chodakova, L., Snahnicanova, Z., Melusova, M., Kozics, K., & Slamenova, D. (2014). Assessment of antioxidative, chelating, and DNA-protective effects of selected essential oil components (eugenol, carvacrol, thymol, borneol, eucalyptol) of plants and intact Rosmarinus officinalis oil. J. Agric. Food Chem., 62: 6632–6639. Hossain, M. A., Al Kalbani, M. S. A., Al Farsi, S. A. J., Weli, A. M., & Al-Riyami, Q. (2014). Comparative study of total phenolics, flavonoids contents and evaluation of antioxidant and antimicrobial activities of different polarities fruits crude extracts of Datura metel L. Asian Pacific Journal of Tropical Disease, 4(5): 378–383.

Khoudja, N. K., Boulekbache-Makhlouf, L., & Madani, K. (2014). Antioxidant capacity of crude extracts and their solvent fractions of selected Algerian Lamiaceae. Ind. Crops Prod., 52: 177– 182. Liepelt, S., Mayland-Quellhorst, E., Lahme, M., & Ziegenhagen, B. (2010).Contrasting geographical patterns of ancient and modern genetic lineages in Mediterranean Abies species.Plant. Syst. Evol. 284: 141–151. NCCLS (2004).National Committee for Clinical Laboratory Standards, Method for Antifungal Disk Diffusion Susceptibility Testing of Yeasts; Approved Guideline. NCCLS document M44-A [ISBN 1-56238-532-1]. NCCLS, 940 West Valley Road, Suite 1400, Wayne, Pennsylvania 190871898 USA. Orak, H. H., Yagar, H., Isbilir, S. S., Demirci, A. Ş.,Gümüş, T., & Ekinci, N. (2011). Evaluation of Antioxidant and Antimicrobial Potential of Strawberry Tree (Arbutus Unedo L.)Leaf. Food Sci. Biotechnol., 20(5): 1249–1256. Poaty, B., Lahlah J., Porqueres, F. & Bouaﬁf, H. (2015).Composition, antimicrobial and antioxidant activities of seven essential oils from the North American boreal forest.World J MicrobiolBiotechnol, 31:907–919. Quezel, P., & Santa, S. (1962). Nouvelle flore de l'Algérie et des régions désertiques méridionale, Tom I, CNRS. Paris, 545 p. Quezel, P. (1985). Les sapins du pourtour méditerranéen, Forêt méditerranéenne, VII(1): 27–34. Quezel, P. (1998). Diversité et répartition des sapins sur le pourtour méditerranéen, Forêt méditerranéenne, XIX(2): 93– 104.

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Rajendran, P., Nandakumar, N., Rengarajan, T., Palaniswami, R., Gnanadhas, E. N., Lakshminarasaiah, U., Gopas, J., & Nishigaki, I. (2014). Antioxidants and human diseases. Clinica ChimicaActa, 436 (25): 332–347. Ramdani, M., Lograda, T., Chalard, P. & Figueredo, G. (2014). Chemical and antimicrobial properties of essential oils of Abies numidica, endemic species of Algeria. International Journal of Phytopharmacology, 5(6): 432–440. Roby, M. H. H., Sarhan, M.A., Selim, K. A.H., & Khalel, K. I. (2013). Antioxidant and antimicrobial activities of essential oil and extracts of fennel (Foeniculum vulgare L.) and chamomile (Matricaria chamomilla L.). Industrial Crops and Products, 44: 437–445.

Source of Support: Algerian Ministry of Higher Education and Scientific Research

Schutt, P., & Lang, Z. U. M. (1991). Tannenarlen Europasund Kleinasiens. Basel; Boston; Berlin: Birkhauser, © 1991 Springer Basel AG, 132 p. DOI: 10.1007/978-3-0348-7689-6 Stefanović, O. D., Tešić, J. D., & Čomić, L. R. (2015). Melilotus albus and Dorycnium herbaceum extracts as source of phenolic compounds and their antimicrobial, antibiofilm, and antioxidant potentials. Journal of Food and Drug Analysis, 23(3): 417–424. Vishnoi, S. P. V., Ghosh, A. K., Debnath, B., Samanta, S., Gayen, S., & Jha, T. (2007). Antibacterial activity of Abies webbiana. Fitoterapia, 78: 153–155. Xia, J.-H., Zhang, S.-D., Li, Y.-L., Wu, L., Zhu, Z.-J., Yang, X.-W., Zeng, H.-W., Li, H.-L., Wang, N., Steinmetz, A., & Zhang, W.-D. (2012). Sesquiterpenoids and triterpenoids from Abiesholo phylla and their bioactivities. Phytochemistry. 74: 178–184.

Conflict of Interest: None Declared

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Research article THE USE OF VARIOUS PLANT TYPES AS MEDICINES BY LOCAL COMMUNITY IN THE ENCLAVE OF THE LORE-LINDU NATIONAL PARK OF CENTRAL SULAWESI, INDONESIA Rosmaniar Gailea1*, Ach. Ariffien Bratawinata2, Ramadhanil Pitopang3, IrawanWijaya Kusuma4 1,2,4

Faculty of Forestry, Mulawarman University, Jl.Ki Hajar Dewantara Kampus Gunung Kelua, Samarinda, Indonesia 75116 3 Faculty of Mathematics and Science, Tadulako University, Jl. Soekarno- Hatta Km 9 Palu 94118 Central Sulawesi , Indonesia. 1 Faculty of Agriculture, Muhammadiyah Palu University, Jl. Hangtuah no 114 Palu 94118 Central Sulawesi, Indonesia. *Corresponding author: gailearos@gmail.com

Received: 14/09/2015; Revised: 09/01/2016; Accepted: 15/01/2016

ABSTRACT The various ethnic communities residing around Lindu Lake in Central Sulawesi Province of Indonesia with their local knowledge on medicinal plants still use them as the source of traditional medication for healing light and serious ailments. Three females and one male key informant who had good reputation in medicinal plant knowledge and traditional healing along with 34 respondents were questioned using semi structured interview method. Ninety six species belonging to 45 families have been found which can be used to heal 87 kinds of ailments, for facial treatment, for post natal care and as food supplements. The medicinal plants can be found in various habitats including house yard, plantation, forest, rice fields, swamps and any other location (wild).

KEY WORDS: Lindu Lake, Lore Lindu National Park, local knowledge, and medicinal plant uses.

Cite this article: Rosmaniar Gailea, Ach. Ariffien Bratawinata, Ramadhanil Pitopang, IrawanWijaya Kusuma (2016), THE USE OF VARIOUS PLANT TYPES AS MEDICINES BY LOCAL COMMUNITY IN THE ENCLAVE OF THE LORE-LINDU NATIONAL PARK OF CENTRAL SULAWESI, INDONESIA, Global J Res. Med. Plants & Indigen. Med., Volume 5(1): 29–40

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INTRODUCTION The traditional use of plants for healing in Indonesia dates back to prehistoric times. The art and knowledge of the uses of plants as medicine have been handed down orally from generation to generation. Some plants still used in traditional medicine can be found depicted in reliefs on the walls of ancient temples in Java. Such as those of Borobudur, Prambanan, Penataran and Sukuh (de Padua et al., 1999) Indonesia has 1,340 ethnics (BPS, 2010) each holds its own knowledge about traditional medication and as a tropical country Indonesia encompass a vast number of biodiversity, many of them exist in Lore-Lindu National Park (LLNT) of Central Sulawesi, Indonesia, which is one of the highly significant conservation regions in Indonesia. Based on several studies, several botanists (Pitopang et al., 2003 and 2004, Mogea, 2002 and 2005, Kessler et al., 2005) have shown that this region is rich in plant types that have their own charm as some of them represent the flora of mountain forest of Sulawesi exhibiting high biodiversity in characteristic and many of them are endemic. However, studies conducted in this region have been limited particularly to those related to the use of medicinal plants by local community. Lindu Lake located in the enclave of LLNT is the eight largest lakes in Sulawesi and the second largest in Central Sulawesi. Community settling around the lake belongs to various ethnics and sub ethnics who still hold the knowledge of traditional medication and use various plants for various illness treatments as well as for health care and cosmetics. The medicinal plants are collected from forest, plantation, house yard, wetland rice fields and swamps existing around their settlement. In general, this research is aimed to expose the local/traditional knowledge and its use for traditional medicines and to record medicinal plants and traditional medication. METHODS a.

Study Area The research area was located around Lindu Lake (Figure 1). This location was selected

based on considerations that it is high in biodiversity and the local community settling in the Lindu lake region comprises of various ethnics. The Lindu enclave recently has become Lindu sub district due to regional expansion of Kulawi sub district by the government of Central Sulawesi Province, Indonesia. Lindu Sub District spread over an area of 58,585.72 ha (BBTNLL, 2012) covering Puroo, Langko, Tomado and Anca villages. Lindu sub district has 78% (45,672.27 ha) of its total area under LLNP and is mountainous at 800–100 elevation above sea level. The slope land is very steep ranging from 60–70% even >80% (BPS, 2013). Climatic seasonality is not pronounced, with a monthly average of over 100 mm of rainfall; although in some years monthly values can be far lower (Whitten et al., 1987). Minimum temperatures range between 12°C and 17°C, while maximum values range from 26°C to 35°C. The natural vegetation in the study area is evergreen tropical forest dominated by the families Anacardiaceae, Burseraceae, Lauraceae, and Sapotaceae (Whitmore & Sayer 1986, Kessler et al., 2005). b. Demographic Social Characteristics of Respondents The number of respondents was 38 persons, 20 years to more than 60 years old residing in four villages. The male respondents were 42.11% and the females were 57.89%. Most of the respondents (84.21%) were farmers who cultivate wetland rice, coffee and cacao while the remaining 18.42% were fishermen who also work as farmers. The respondents living in the enclave of Lindu Lake were predominantly Kaili ethnic i.e Kaili Tado sub ethnic. The Kaili ethnic who settled in Lindu region are subdivided into 7 different sub ethnics each with its own dialect including Kaili Ledo, Kaili Ija, Kaili Ado, Kaili Moma, Kaili Tohulu, Kaili Uma, and resettled Kaili Da’a (BTNLL, 2009). Other ethnic outside Kaili ethnic are Pekuehua, Besoa and Bada (BTNLL, 2009). Some ethnics coming from outside Central Sulawesi are Bugis, Jawa, Minahasa and Toraja (Figure 2).

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Figure 1a. Map of Central Sulawesi

Figure 1b. Map of Lore Lindu National Park, Indonesia

Figure 2. Ethnic Composition residing in Lindu Enclave, Indonesia Minahasa Arab 3% 6% Jawa 7%

Lonca 10% Bugis 13%

Data Collection

This research was conducted through interview for ethnopharmachology data collection and through laboratorial analysis for determination of chemical characteristics and

Kaili Tado 61%

bioactivity of medicinal plants from April 2013â&#x20AC;&#x201C;July 2014. Four key informants (three females and one male) from each village were selected based on their good reputation in knowledge of medicinal plants and traditional medication and 34 respondents who sufficiently

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have the knowledge were also chosen. Semi structured interview were performed with the key informants and the respondents. The collection of medicinal plants for herbarium and determination of habitat were done directly in field through survey method (Balgooy, 1987). The medicinal plants found were collected and made for herbarium specimens. The identification of the medicinal plants was done in the Celebences Herbarium of Tadulako University and the Biological Research Centre of The Indonesian Institution of Sciences (LIPI) Bogor.

and Fabaceae 5 species each, Araceae and Euphorbiaceae 4 species each and the other families are present only less than 4 species each.

RESULTS

The most common and simplest way of processing the medicinal plants are by drinking the decoction made by boiling the plants (75%). The plants were also used in such a way that they are applied to body parts (32%). Whilst other modes of administration range between 1â&#x20AC;&#x201C;7% (Figure 3).

Medicinal Plant Community

Types

Used

The number of medicinal plants found in the research area was 96 species used by the community around Lindu lake. The knowledge of these medicinal plants and its uses has been passed on from their ancestors to the present generation. The medicinal plants can be grouped into 45 families. Asteraceae is the dominant family comprised of 9 species, followed by Lamiaceae 8 species, Piperaceae

2. Use and Processing of Medicinal Plants as Traditional Medications Based on the local knowledge, the 98 medicinal plants are useful to treat 87 types of light to serious ailments. The plants also can be used for cosmetics, antiseptic, tonic, post natal treatment, etc.

The part of plant that was mostly used by the community is the leaves (60%) followed by the whole plant (16%), and the other parts (1â&#x20AC;&#x201C; 5%) (Figure 4).

Figure 3. The various kinds of the medicinal plant preparation 60 50

Percent use

40 30 20 10 0 Decoction

Affixed

Juice

Eaten

Mode of preparation

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Sorted

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Figure 4. The parts of the medicinal plants used for treating ailments 70

Percent use

60 50 40 30 20 10 0

Plant part

Table 1. Ethnomedicinal uses of medicinal plants of Lindu Lake Vr. no. 1

Scientific name (family)

Local name

Part used Old leaves Leaves

Mode of Medical uses preparation Decoction Tuberculosis, difficulty urinating Decoction Fever

Allamanda cathartica L.(Apocynaceae) Cordia corymbosa Miq. (Boraginaceae) Jatropha curcas L. (Euphorbiaceae)

Alamanda

Balacai

Leaves, stem, sap

Affixed, juice

Bambusa sp. (Poaceae)

Bambu batu

Bud

Amaranthus spinosus L. (Amaranthaceae) Scurrulaarto purpurea (Blume) Danser. (Loranthaceae);

Bayam duri

Leaves

Eat it in its raw state Decoction

Benalu kakao, Benalu kopi, Benalu pohon lemon Bila

Whole plant

Decoction

Tumor, blood vomiting

Leaves

Decoction

Bolubua

Leaves

Decoction

Bolukaruke

Leaves

Decoction

Hernia, Osteoporosis Body odor, Bad breath Epistaxis

Bolulasu

Leaves

Affixed

Body odor

Bolutana

Leaves

Decoction

Body odor

2 3

7 8 9 10 11

Crecentia cujeta L. (Bignoniaceae) Piper betle L. (Piperaceae) Piper retrofractum Vahl. (Piperaceae) Piper majusculum Blume. (Piperaceae) Piper sp. (Piperaceae)

Ambarogo

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Lack of appetite, Cough, skin allergies on children, blood vomiting, headache, thrush Hepatitis, Diabetes Constipation

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Clerodendron japonicum (Thumb.) Swett.(Lamiaceae) Selaginella sp.(Selaginellaceae) Amydrium zippelianum (Schott) Nicolson (Araceae) Plantago major L. (Plantaginaceae)

Bunga pagoda Leaves

Decoction

Ulcer

Cakarayam

Decoction

Rheumatism

Daun rusuk

Whole plant Leaves

Affixed

Sore ribs

Daun sendok

Leaves

Affixed

Gamal

Leaves

Decoction

Gedi

Leaves

Decoction

Gliricidia sepium (Jacq.) Walp.(Fabaceae) Abelmoschus manihot (L.) Medik (Malvaceae) Clerodendrum sp.(Lamiaceae)

Diseases of the descending colon, appendix Headache

Gonato

Leaves

Decoction

Scutellaria sp. (Lamiaceae)

Hipodo Langko

Whole plant

Decoction

Curangafel-ferrae (Lour.) Merr. (Linderniaceae) Psidium guajava L. (Myrtaceae) Citrus maxima (Burm) Merr. (Rutaceae) Hibiscus tiliaceus L.(Malvaceae)

Hipodo Walo

Decoction

Jambu batu

Whole plant Leaves

Jeruk bali

Leaves

Decoction

Kalebou

Shoot

Decoction

Cordia myxa L (Boraginaceae) Apium graveolens L. (Apiaceae) Ocimum basilicum L. (Lamiaceae) Senna alata (L.) Roxb. (Fabaceae) Syngonium podophyllum (Araceae) Moringa oleifera Lam. (Moringaceae) Spilanthes oleracea L.(Asteraceae) Coffea canephora Pierre ex A.Froehner (Rubiaceae) Kalanchoe pinnata Pers (Crassulaceae) Orthosiphon aristatus (Blume) Miq. (Lamiaceae)

Kanuna

Branch

Juice

Kaporontomate

Decoction

Kayu manuru

Whole plant Leaves, stem Leaves

Keladi bunga

12 13 14

16 17

21 22 23

24 25 26 27 28 29 30 31 32 33

Decoction

Pain related to Kidney ailments Back pain, worms, postpartum Stomach pain, Acute internal disease Internal disease Diarrhea, Strengthen teeth Diabetes Fossilized stomach, hemorrhoid Postpartum, Fever

Juice

Persistent cough more than100 days Fever and cough

Juice

Epilepsy, sore skin

Leaves

Decoction

Antidote

Kelor

Leaves

Decoction

Lowers body heat

Kondo uwe

Flower

Juice

Tooth ache

Kopi arabika

Leaves

Decoction

Cosmetics

Kujadi

Leaves

Juice

Kumis kucing

Leaves and stem

Decoction

Headache, cough, high fever, blain Back pain

Kapumpu

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36 37 38 39

41 42 43 44 45 46 47

48 49 50 51 52 53 54

Zingiber zerumbet (L.) Roscoe ex. Sm. (Zingiberaceae) Cucurbita moschata Duchesne ex Poir (Cucurbitaceae) Nicotiana tobacum L. (Solanaceae) Elephantopus mollis Kunth.(A steraceae) Glinus oppositifolius (L.) Aug.DC. (Molluginaceae) Drymaria cordata (L.) Willd. ex Schult. (Caryophyllaceae)

Kurondo

Rhizome

Sorted

Elephant foot pain

Labu kuning

Leaves

Affixed

Eye disease

Lamba

Leaves

Affixed

Strengthen teeth

Lambantomate

Whole plant Whole plant Whole plant

Eat it in its raw state Decoction

Stroke, Tooth ache, postpartum Renal stones

Juice

Amaranthus blitum subsp. oler aceus (L.) Costea (Amaranthaceae) Pollia secundiflora (Blume) Bakh.f. (Commmelinaceae) Pogostemon auricularius (L.) Hassk.(Lamiaceae) Alstonia scholaris (L.) R. Br. (Apocynaceae) Ficus septica Burm.f. (Moraceae) Macaranga hispida (Blume) MĂźll.Arg. (Euporbiaceae) Cassia sp.(Fabaceae) Plectranthus scutellarioides (L.) R.Br. (Lamiaceae)

Lasuani

Leaves

Decoction, Affixed

Epilepsy, headache, diabetes, high heat in infants Headache

Lekosa

Affixed

Lengaru

Leaves and stem Leaves and stem Bark

Decoction

Leg and stomach swelling Persistent cough more than100 days Wound infection

Levonu

Shoot

Juice

Eye disease

Mapo

Fruit

Affixed

Remove warts

Mate sambula Mayana

leaves Leaves

Decoction Juice

Bryophyllum pinnatum (Lam.) Oken (Crassulaceae) Synedrella nodiflora (L.) Gaertn. (Asteraceae) Dichrocephala integrifolia (L. f.) Kuntze (Asteraceae) Hyptis suaveolens (L.) Poit. (Lamiaceae) Sambucus javanica Reinw. ex Blume (Adoxaceae) Hemigraphis sp.(Acanthaceae) Begonia aptera Blume (Begoniaceae)

Mpomata

Leaves

Juice

Epilepsy Contusion, internal disease, ulcer, cough Carbuncles

Anjing gila

Leaves

Keteguran

Leaves

Juice/Decoct Stray dog bites ion Juice Worm infestation

Kutu air

Leaves

Juice

Water fleas

Obat jerawat

Fruit

Affixed

Acne medication

Bunga merah hijau Nura

Leaves

Decoction

Ulcer, lochia

Branch

Juice

Worm infestation

Langalo Languntule

Lelompeba

Decoction

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55 56 57 58 59 60 61

62 63 64 65 66

67 68

69 70 71 72 73 74 75

Eleusineindica (L.) Gaertn(Poaceae) Luffa cylindrica L.(Cucurbitaceae) Scindapsus pictus Hassk. (Araceae) Sporobolus diandrus (Retz.) P.Beauv.(Poaceae) Merremia umbellata (L.) Hallier f. (Convolvulaceae) Centella asiatica (L.) Urb.(Apiaceae) Phyllanthus urinaria L. (Phyllanthaceae)

Pada Palolaboe

Young leaves Dry fruit

Affixed/Dec oction Decoction

Pancihinana

Leaves

Affixed

Abdominal swelling Tinea versicolor

Pancisilanalida

Leaves and stem Leaves

Affixed

Black spot

Affixed

Black spot

Pancongkolangi

Leaves

Decoction

Hacking cough

Panuntu

Whole plant

Decoction

Cyperus killingia Endl. (Cyperaceae) Ageratum conyzoides L.(Asteraceae) Solanum lycopersicum L. (Solanaceae) Momordica charantia L. (Cucurbitaceae) Hippochaete debilis (Roxb. ex Vaucher) Ching.(Equisetaceae) Jatropha multifida L. (Euphor biaceae) Eleutherine bulbosa (Mill.) Urb.(Iridaceae)

Paparisipa

Leaves

Affixed

Ulcer, liver & kidney ailments, Smooth bowel movement Fever

Paralente

Leaves

Juice

Parancina

Leaves

Affixed

Paria

Leaves

Juice

Anredera cordifolia (Ten.) Steenis(Basellaceae) Etlingera elatior (Jack) R.M.Sm.. (Zingiberaceae) Eclipta prostata (L.) L. (Asteraceae) Mimosa pudica L. (Fabaceae) ; Amphineuron sp. (Thelypteridaceae) Murdannia blumei (Hassk.). (Commelinaceae) Myrmecodia sp. (Rubiaceae)

Pancisilana lore

Injury

Hard to breathe, ulcer, liver disease Burns, reduce heat

Pasolonteneru Stem

Sorted

Reduce heat, cough Fracture, twist

Penesilin

Sap

Afficed

Bleeding, burns

Piatopoule

Tuber

Decoction

Pinahong

Leaves

Decoction

Posuntikala

Flower

Decoction

Puro

Juice

Sakimalei

Whole plant Whole plant Leaves

Sampularo

Leaves

Affixed

Sarangsemut

Bulbus

Decoction

Heart disease, migraine, hemorrhoid Tumor, high blood pressure Hyperuricemia, food Umbilical cord injuries, thrush Insomnia, goiter, heat in, appendix Hot and red spots on the body Black spots on the face Ulcer, headache high blood pressure, hemorrhoids

Putri malu

Decoction Decoction

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Sida rhombifolia L. (Malvaceae) Euphorbia hirta L. (Euphorbiaceae) Melastoma malabathricum L. (Melastomaceae) Annona muricata L.(Annonaceae)

Silaguri

Leaves

Decoction

Simulasi

Decoction

Sinduru

Root and leaves Leaves

Sirsak

Leaves

Decoction

Bidens pilosa L.(Asteraceae) ;

Susupi

Leaves

Affixed

Cordyline fruticosa (L.) A.Chev. (Asparagaceae) Hyptis capitata Jacq. (Lamiaceae) Tambajara

Taba

Leaves

affixed

Taba

Leaves, flower, root

Decoction

Sennatora L.(Roxb.) (Fabaceae) Physalis angulata L.(Solanaceae) Scleria purpurascens Steud. (Cyperaceae) Piper umbellatumL. (Piperaceae)

Tambuangkebe

Root

Decoction

Tampei

Leaves

Decoction

Tatari

Shoot

Decoction

Tavalevo

Leaves

Commelina diffusa Burm.f. (Commelinaceae) Rhapidophora sp.(Araceae) Ipomea batatas (L.) Poir (Convolvulaceae) Cheilocostusspeciosus (Costaceae) Angiopteris evecta (Marattiaceae) Poikilospermum suaveolens (Bl.) Merr (Urticaceae) Anthocepalus chinensis (Rubiaceae) Eleocharis artopurpurea Retz. Presl.(Cyperaceae) Crassocephalum crepidioides (Benth) (Asteraceae) Erigeron sumatrensis Retz. (Asteraceae)

Tavelehoka

Leaves

Decoction/A Ailments of the ffixed descending colon, pain in the penis, hemorrhoid. Affixed Cramps, gout

Tavemolu Tavovi

Affixed Decoction

Back pain arise on the body

Tintiase

Leaves Young Leaves Stem

Juice

Fever

Tombila

Stem

Juice

Stray dog bites

Tombu

Leaves, stem Bark

Decoction Decoction

Breast cancer, eye pain, Malaria

Affixed

Postpartum

Wavaro

Whole plant Leaves

Affixed

Cuts

Wingkotu

Leaves

Affixed

Leprosy

76 77 78 79

83 84 85 86

87 88 89 90 91 92 93 94 95 96

Towote Voluntile

Decoction

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Wound infection, strengthen teeth Ailments of Appendix Boil, epistaxis Renal stone, high blood pressure, wounds Post partum, cough Sore neck Renal stones, liver disease, wounds, cough, shortness of breath, diabetes Blood mixed stools High blood pressure, migraine Renal stones

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3. Habitat of Medicinal Plant Found by Community Habitat of the medicinal plants found by the community is spread over from the edge of the lake to the forest around Lindu (LLNP).

The types of the habitat depicted in Figure 5 include secondary forest, house yard, wetland rice fields, swamps, and wild (any other location).

Figure 5. Habitat of the medicinal plants found 35.00

Percent use

30.00 25.00 20.00 15.00 10.00 5.00 0.00 Forest

Garden

Yard

Wet rice field

Swamp

Wild

Habitat

The house yard is the most common habitat where the medicinal plants exist. Multifunctional community yards around Lindu Lake have diverse vegetation/plants including ornamental plants, vegetable and such plantation crops as Coffea robusta / arabica, Theobroma cacao, Psidium guajava, etc. The medicinal plants cultivated are generally Jatropha curcas, Cheilocostus speciosus, Ocimum basilicum and Eleutherine bulbosa while others are allowed to grow wildly such as Plantago major, Crassocephalum crepidioides, Hyptis caitatpa, etc.

purpurelens, Comelina Rhapidophora sp.

Plantation very often exists next to either secondary forest or primary forest where Coffea arabica/robusta and Theobroma cacao are mainly cultivated. Other vegetation are also found in the plantation like Amaranthus spinosus, Elephantopus mollis, Drymaria cordata, Amaranthus lividus, etc.

DISCUSSION

Some vegetation often found in the forest is Polia secundifolia, Alstonia scholaris, Scindapsus pictus, Merremiaum bellata, Amphineuron sp, Myrmecodia sp., Scleria

diffusa

and

Several vegetation grow in various habitat frequently in open land, road side, and gutter edge. This vegetation includes Amaranthus spinosus, Elephantopus mollis, Drymaria cordata, Amaranthus lividus, etc. Centella asiatica, Sporobulus diandrus, Murdannia blumei, etc, are easily found in wetland rice fields whereas Zingiber zerumbet and Eleocharis artopurpurea were found in swamp areas.

The local community dwelling around Lindu Lake has been keeping their knowledge on different kinds of plants grown around their settlement which can treat various ailments, health care and cosmetic from generation to generation. Although health services provided by government have increasingly improved, the tradition to treat illnesses and health care using medicinal plants are still being frequently practiced. Of 38 respondents, 72.68% were between 20â&#x20AC;&#x201C;50 years old and the remaining

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26.32% over 60 years old. This indicates that the knowledge of the medicinal plants and their uses is still being maintained and practiced in the daily life of the Lindu lake community. In this research, the dominant medicinal plant family found is Asteraceae. Previous researches showed that the Asteraceae family is relatively wide spread compared to the other families (Marles and Farnsworth, 1995 and Trojan et al., 2011). Giulietti et al. (2005) placed the family of Asteraceae and Myrtaceae as those which exhibit their abundance in high biodiversity and have rapid secondary metabolical development. The part of the medicinal plant that is mostly used by the community around Lindu lake is leaves (60%). Several researches showed that leaves are plant parts that are mostly used for traditional medication (Kassam, 2011; Packera, 2012; Moushumi, 2013; Deepak and Gopal, 2014; Laid, 2014). From the view of conservation, the use of leaves as source of traditional medicine has no harmful effect on the environment. However, efforts to cultivate the medicinal plants where leaves are frequently used for medicine are necessary to maintain their sustainability and easy access. The Lindu lake community has not yet cultivated medicinal plants used frequently because they can be found easily around their settlement. The most common way to process the parts of the medicinal plants is by boiling them in water (57%) (Figure 3). The processed medicinal plants are generally classified into complex concoction and single concoction

made up by various type of plants and by only one plant, respectively. The people generally boil the medicinal raw materials with water allowing half of the initial water amount to be left and the water extract is administered for drinking. The leaves used are odd in number such as 3, 5, 7 leaves, etc. Rhizome used usually ranges from one to several segments. The existence of different kinds of the medicinal plants in their habitat is important to be identified because the ecological aspect of the plants can uncover their actual condition. Such ecological data as biology, habitat and others is significantly important to future development of the medicinal plants. CONCLUSION A total of 96 species belong to 45 families were found as medicinal plants used by community residing around Lindu Lake (The enclave of Lore Lindu National Park) to treat different kind of ailments and other uses such as facial health care, post natal care and food supplement. The local knowledge of medicinal plants and traditional healing has been handed down from generation to generation. ACKNOWLEDGEMENT This research was funded by Research and Community Service Directorate of Higher Degree General Directorate of Education and Culture Ministry of Indonesia through Doctor Dissertation Funding in 2014. We thanks the people of Lindu lake whose helps made this research possible.

REFERENCES Balgooy van MMJ (1987). Collecting In: de Vogel (ed) Manual of Herbarium Taxonomy: Theory and Practice. UNESCO. De Padua LS, Bunyapraphatsara N, Lemmens RHMJ (ed) (1999). Plant Resources of South-East Asia. 12 (1) Medicinal and poisonous plants 1. Prosea Bogor Indonesia

Deepak P, Gopal GV (2014). Ethnomedicinal Practices of Kurumba Tribes Niligiri District Tamil Nadu India in Treating Skin Diseases. Global Journal Res. Med. Plants & Indegenous Medicine 3(4):8â&#x20AC;&#x201C;16.

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Giulietti AM, Harley RM, Queiroz LP, Wanderley MGL, Van Den Berg C (2005). Biodiversity and conservation of plants in Brazil. Conservation Biology 19:632–663. Indonesia Central Bureau of Statistics (BPS) (2010). Indonesia Sub District in Figures Indonesia Central Bureau of Statistics (BPS) (2013). Kulawi Sub District in Figures. Kassam KA, Karamkhudoeva M, Ruelle M, Baumflek M (2010). Medicinal Plant Use and Health Sovereighty: Findings from the Tajik and afghan Pamirs. Hum. Ecol. 38: 817–829 Kessler M, Kessler PJA, Gradstein SR, Bach K, Schmull M, Pitopang R (2005). Tree diversity in primary forest and different land use systems in Central Sulawesi, Indonesia. Biodiversity Conserv. 14:547–560 Lore Lindu National Park Office (BTNLL) (2010). Potential Areas of Lindu Lore Lindu National Park Office (BTNLL) (2012). GIS data of Lore Lindu National Park.

International Symposium” The Stability of Tropical Rainforest Margins. Linking Ecological, Economic and Social Constrains of Land Use and Concervations” Georg-AugustUniversity of Gottingen, September 19– 23 Packera J, Brouwera N, Harringtona D, Gaikwada J, Heron R, Ranganathana S, Vemulpada S, Jamiea J. (2012) An ethnobotanical study of medicinal plants used by the Yaegl Aboriginal community in northern New South Wales, Australia. Journal of Ethnopharmacology 139: 244–255. Pitopang R, Gradstein SR (2003). Herbarium Celebence (CEB) and Its Role in Supporting research on Plant Diversity of Sulawesi. Biodiversitas 5: 36–41 Pitopang R, Kessler M, Kessler PJA, Gradstein SR (2004). Tree Diversity in Primary Forest, Secondary Forest and Forest Garden in Central Sulawesi, Indonesia. Abstract, Sixth International Flora Malesiana Symposium, Los Banos, Philippines, 20–24

Marles R, Farnsworth N (1995). Antidiabetic plants and their active constituents. Phytomedicine 2: 137–165.

Trojan-Rodriguesa M, Alvesa TLS, Soaresa GLG, Ritter MR (2011). Plants used as antidiabetics in popular medicine in Rio Grande do Sul, southern Brazil. Journal of Ethnopharmacology 139:155–163

Mogea JP (2002). Preliminary Study on the Palm Flora of the Lore Lindu National Park Central Sulawesi Indonesia. Biotropia 18:1–20

Whitmore TC, Sayer JA (1986). Composition and structure of a lowland rain forest at Toraut, Northern Sulawesi. Kew Bull. 41:747–757.

Mogea JP (2005a). Diversity and Density Palms and Rattans in Primary Forest, Old Secondary Forest, and Recent Estabilished Traditional Cacao and Coffee Garden in Central Sulawesi, Indonesia. Abstract: Proceedings

Whitten AJ, Muslim M, Henderson GS (1987). The Ecology of Sulawesi. GadjahMada University Press, Yogyakarta, Indonesia

Source of Support: Research and Community Service Directorate of Higher Degree General Directorate of Education and Culture Ministry of Indonesia

Conflict of Interest: None Declared

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Case Study A CASE STUDY ON THE AYURVEDIC MANAGEMENT OF VARICOSE VEIN Shona Rudolph1*, Prasanth D2, Ananthram Sharma3 1

Second Year P.G Scholar, Dept. Of P.G studies in Panchakarma, Amrita School Of Ayurveda, Vallickavu, Kollam. Dt., Kerala, India 2 Assistant Professor, Post Graduate Department of Panchakarma, Amrita School of Ayurveda, Vallickavu, Kollam Dt., Kerala, India 3 Associate Professor, Post Graduate Department Of Panchakarma, Amrita School Of Ayurveda, Vallickavu, Kollam Dt., Kerala, India *Corresponding Author: E-mail: shonamagdaline@gmail.com; Mobile no: 09747035400

Received: 05/11/2015; Revised: 11/01/2016; Accepted: 20/01/2016

ABSTRACT A condition, in which a vein becomes dilated, elongated and tortuous, this is said to be “Varicose”. There are primary as well as secondary causes of varicose veins. The superficial perforating as well as the deep veins are involved in varicose veins. The contemporary treatment for varicose veins includes ligation, ligation with stripping surgical treatments. But these treatments can cause reoccurrence of this disease. Ayurveda texts reveal Siravyadha (venesection) cures varicose veins. We hereby report a case of a 60 year old female with complaints of pain in the lower limbs from calf to the dorsum of the foot of both legs associated with dilated and tortuous veins in the anterior aspect of lower leg with discolouration. She was advised admission for ten days and was treated with internal medication, external treatments and Siravyadha (venesection). During the treatment all the signs and symptoms of varicosity reduced to a very high extend. KEYWORDS: Varicose vein, Siragranthi, Siravyadha, Ayurveda

Cite this article: Shona Rudolph, Prasanth D, Ananthram Sharma (2016), A CASE STUDY ON THE AYURVEDIC MANAGEMENT OF VARICOSE VEIN, Global J Res. Med. Plants & Indigen. Med., Volume 5(1): 41–48

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INTRODUCTION: The common sites of varicosity are superficial venous system of the lower limbs, it affects either the long or short saphenous vein or both (Das.S, 2014). The superficial veins and perforating veins, connects the superficial with deep veins. The primary causes of varicose veins are the incompetency of the valves and weakness in the walls of veins which causes venous insufficiency. Secondary varicose veins occur due to venous obstruction caused by pregnancy, tumours in the pelvis, fibroid, ovarian cyst and deep vein thrombosis etc. (Das, 2014). In Ayurveda classics varicose vein can be very much co related with the signs and symptoms explained for Siragranthi (obstructive circulation). Due to Vataprakopaka nidanas (causative factors which increases vata) such as, physical exertion, straining, for debilitated persons the vitiated vata enters the Siras (veins) causing the Sampeedana (Squeezing), Sankocha (constriction), and Vishoshana (act of drying up) which produces round and protruded Granthi (cyst) in the Siras (Veins), manifesting Siragranthi (varicose vein) (Acharya YT, 2009). Hence Ayurveda reveals Siravyadham (venesection) cures Siragranthi (varicose veins) (Harisastri Paradakara, 2011). In this article a case study in varicose vein has been taken for understanding the releaveness of complaints presented by a 60year old woman who has visited the out patient department. MATERIALS & METHODS A 60 year old woman visited the O.P.D of Amrita school of Ayurveda, Kerala, India on 909-2015 with M.R.D no: 65455 was advised admission for ten days and got discharged on 18-09-2015. She presented with complaints of pain in the lower limbs from calf to the dorsum of foot of both legs and had prominent, dilated, tortuous veins at the antero medial aspect of the lower 1/3rd part of the leg as well as postero lateral aspect of the calf region of left leg since 15 years (Figure 1). She also had complaints of difficulty in passing stool and an irregular

bowel. She had a dull aching pain associated with throbbing sensation in the foreleg and ankle region of the left leg as well as burning sensation on both soles since 3 months. The pain aggravated on long standing especially in the evenings and night hours accompanied with slight swelling at the left ankle region, since 3 months. On examination, there was pain, swelling and dilated tortuous veins present in the foreleg as well as in the calf region, mild swelling seen in the left ankle region, slight bluish discolouration was evident. No ulceration was noticed. Pain got relieved by elevation of legs. Moseâ&#x20AC;&#x2122;s Sign (Das.S, 2014) (pain in the calf region on gentle squeezing of calf region) was slightly positive in the patient. The patient was subjected for Siravyadham (venesection) on alternate days along with Sarvanga Abhyanga (full body massage), Lepa (application of medicated paste) as well as with Pitta Samana Aushadi (medicines which pacifies pitta) internally. Treatment protocol: On the day of admission internal medication was adopted using Amruthotharam Kashayam (Krishnan K.V, 2006) and Vasaguluchyadi Kashayam (Krishnan K.V) was advised on a dose of 20 ml Kashayam with 60ml warm water at 6.30 in the morning and 6.30 in the evening. Kaishora Guggulu (Krishnan K.V). 1tablet in the morning and 1 tablet in the evening along with Kashayam was administered along with Sooranavaleham (Vaidya Harisastri, 2011) 1 teaspoon at bed time was administered. Sarvanga Abhyanga (full body massage) was done with Pinda tailam (Krishnan.K.V, 2006) for seven days. From the next day onwards Lepam (application of medicated paste) with Gruhadhoomadi Choorna (Vaidya Harisastri, 2011) mixed with hot water was applied on both the legs from below knee. Siravyadham (venesection) was opted to be done on alternate days. Like this three Siravyadham (venesection) was done within the ten days of admission. On the first day of Siravyadham (venesection) 100 ml of blood was removed. On the second day of Siravyadham (venesection) 90 ml of blood was

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removed and on the last day of Siravyadham (venesection) 80 ml of blood was removed. On discharge she was administered with Mahamanjishtadi Kashayam (Krishnan.K.V, 2006) on a dose of 20 ml kashayam and 60 ml of warm water, morning and evening, Kaishora Guggulu (Krishnan.K.V, 2006) 1 tablet in the morning, afternoon, and evening was also given along. Madhuyashtyadi tailam (Vaidya Harisatri, 2011) was administered for external application. The ingredient details of

these formulations have been presented in table 1. The overall reduction in Pain, burning sensation, swelling, tortuosity & skin changes were graded based on patient’s presentation & physician’s observation & were manually documented. The mode of gradation & assessment is explained in Table 2 & 3.

FIGURE – 1: Dilated tortuous vein before treatment.

TABLE – 1: Key Ingredients & Indications of the medicaments prescribed: SL. no 1

Drug Name Pinda Tailam

Amruthotharam kashayam

Vasaguluchyadi kashayam

Key Ingredients Madhuchishta (Bees wax), Manjishta (Rubia cordifolia), Sarjarasa (Vateria indica), Sariva (Hemidesmus indicus) Nagara (Zingiber officinale), Ativisha (Aconitum heterophyllum), Musta (Cyperus rotundus), Bhunimba (Andrographis paniculata), Amrita (Tinospora cordifolia), Vatsaka (Holarrhena antidysentrica) Vasa (Adathoda vasica), Guduchi (Tinospora cordifolia), Aragvadha-bark (Cassia fistula)

Indications Vataraktam (gout)

Puranajwara (prolonged fever), Vatarakta (gout), Amapachana, kamala (jaundice).

Pandu (anemia), Raktapittam , Kamala (jaundice)

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Mahamanjishtadi Kashayam

Kaishora Guggulu

Sooranavaleham

Gruhadhoomadi Choornam

Madhuyashtyadi Tailam

Manjishta (Rubia cordifolia), Musta, (Cyperus rotundus) kutaja (Holarrhena antidycentrica), Guduchi, (Tinospora cordifolia), kushta (Saussurea lappa), Nagara, (Zingiber officinale), Bharangi (Clerodendrum indicum) Vacha,(Acorus calamus), Chitraka (Plumbago zeylanica), Satavari (Asparagus racemosus), Trayamana (Gentiana kurroa), Pippali (Piper longum), Indrayava (Holarrhena antidysentrica), Vasa (Adhatoda vasica) Triphala, Guggulu (Comiphora mukul), Guduchi (Tinospora cordifolia), Vidanga (Embelia ribes), Danti (Baliospermum montanum),Trivrit (Operculina turpethum) Soorana (Amorphophallus campanulatus), Chitraka (Plumbago zeylanica), Nagara (Zingiber officinale), Maricha (Piper nigrum), Guda (Jaggery) Gruhadhooma, Vacha (Acorus calamus), Kushta (Saussurea lappa), Haridra (Curcuma longa) Madhuyasti (Glycyrrhiza glabra), Tamalaki (Phyllanuthus niruri), Durva (Cynodon dactylon), Payasya (Pueraria tuberose), Chandana (Santalum album), Loha, Madhuparni, Satapushpa (Anethum sowa), Jivanti (Leptadenia reticulate)

Vatarakta (gout), Supti (numbness), Ardida (facial paralysis)

Vatasonita (gout)

Mudhavata (obstructed vata), Malabandha (constipation), Mandagni (less digestive fire) Vatashonita (gout), Kaphaja Vatarakta Vatashonita, Pittavikara, Daha (burning sensation), Arttii

TABLE -2: Assessment parameters adopted - Subjective 1. Shoola (Pain) Absent No Pain Mild Occasional pain after long exertion Moderate Frequent pain Severe Pain throughout the day

0 1 2 3

0 1

2. Daha (Burning sensation): Absent Present

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Global J Res. Med. Plants & Indigen. Med. | Volume 5, Issue 1 | January 2016 | 41â&#x20AC;&#x201C;48

TABLE-3: Assessment parameters adopted - Objective 1. Shotha (Swelling): Absent Present

0 1

2. Tortuosity 0 1 2 3

Absent Mild Moderate Severe

No dilated veins Few veins dilated after exertion Multiple veins confined to calf or thigh Extensive involving both calf and thigh 3. Skin changes

0 1 2

Absent Mild Moderate

No discolouration Blackish patchy hyper pigmentation Hyper pigmentation with eczema

RESULTS AND DISCUSSIONS: During the ten days of treatment patient underwent a gradual relief in the signs and symptoms. On the day of admission (9-092015) the treatment started with Sarvanga Abhyanga (full body massage) and Lepam (application of medicated paste). From the third

day patient felt slight relief in Shoola (pain) as well as Daha (Burning sensation).The gradations of the symptoms in days during treatment days are shown in tables 4 and 5. There was significant reduction in the signs and symptoms on consideration with the VAS pain scale too (Figure 2).

TABLE-4: Assessment chart - subjective parameters

Shoola (pain) Daha (Burning sensation)

DAY-2 BT AT 3 2 1 1

DAY - 4 BT AT 2 1 1 0

DAY -7 BT AT 1 0 0 0

TABLE -5: Assessment chart - objective parameters

Shotham (oedema) Tortuosity Skin Changes

BT 1 3 2

DAY -2 DT AT 1 1 2 2 1 1

DAY - 4 BT DT AT 1 0 0 2 1 1 1 1 1

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BT 0 1 1

DAY -7 DT 0 1 1

AT 0 1 0

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FIGURE-2 Vas pain scale 10

VAS Scale

PAIN SCALE

8 6 4 2 0 1

NO OF DAYS OF TREATMENT

VAS pain scale - This is said as visual analog scale. Here the gradation of pain has been figured during the ten days of treatment.

The internal medicines given were primarily aimed in pitta shamana (pacifies pitta) and Raktaprasadaka (blood purifying) thereby clearing the underlying pathology of Siragranthi. Sooranavaleham was prescribed for KoshtaShuddhi (clearing the bowels) as a pathway for Vatanulomana (normal movement of vata). Siravyadham (venesection), is the treatment of choice prescribed by the classics for Siragata granthi (Varicose Vein) by Acharya Vagbhatta (Vaidya Harishastri, 2011) and (Acharya Sharangadhara Parashuram Sastri, 2006) considers Siravyadham (venesection) as Ardhachikitsa (half treatment) of Shalya Tantra (Surgery) and is said to provide immediate results than Snehadi Karmas (regular medications) (Acharya YT, 2009). The accumulation of Rakta (blood) and vitiation of vata in the Siras (veins) leads to dilation of veins and tortuosity. Repeated Siravyadham (venesection) brings down the local Shotha (swelling) local congestion. This promotes proper circulation where the stasis is cleared off. During the second phase, Daha (burning sensation) which is one the feature of Pitta is also relieved by Siravyadham (venesection) procedure. This could have been due to the reduced the localised intravascular pressure. Tortuosity was partially relieved in this study may be due to the regaining of normal elasticity by external punctures and the expulsion of static blood. The procedure of Siravyadham (venesection) done is briefly explained under three titles.

Poorvakarma (Pre operative procedure): The following materials were collected prior to the procedure, gauze piece, swabs, and bandages, tourniquet, kidney trays, ounce glass, beaker, scalp vein (no.20), spirit, chairs and dressing table. The leg to be punctured was flexed and a torniquet was tied just above the calf region to visualize the veins carefully after Abhyanga (localized oil massage) and Nadi Swedana (fomentation). Pradhana karma (Main procedure): The most tortuous vein was selected and punctured with no.20 Scalp vein set (butterfly canula), carefully the blood was observed and let flow on to a placed beaker. Later, the output was measured with the help of ounce glass. Paschat karma (Post-operative Procedure): After completion of the procedure, the flow of the blood decreased and finally ceased; after which the needle was withdrawn. The pricked part was cleaned up with cotton swab and tight bandaging was done along with Haridra powder (Curcuma longa) and Gritha (cowâ&#x20AC;&#x2122;s ghee). Later the patient was advised rest by raising her legs using a pillow. The figures (Figure 3, 4, 5) below depict the differences in the Varicosed veins and discolouration during Siravyadham procedure on alternate days.

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FIGURE -3

During Siravyadham on the first day with bluish discoloured,dilated veins

FIGURE -4

On the second day of Siravyadham still the dialated,discoloured veins are present.

There was a gradual relief in the signs and symptoms during this treatment course. The result obtained after the external treatments and internal medication was encouraging. The results were improved at the end of 10th day. Pain, swelling, burning sensation were reduced considerably. Dilated and tortuous veins in the lower leg have reduced in size. Bluish discolouration which was present in the dilated veins, reduced considerably. Siravyadham (venesection) along with internal medication resulted the reduction of signs and symptoms of Siragranthi (varicose veins) specially the symptoms like Shoola (pain), Daha (burning sensation) and Vaivarnya (discolouration). After the follow up period of two weeks there was no aggravation in the disease. The treatments employed were purely based on the principles of Ayurveda and was found to be very effective. The condition of the patient improved remarkably.

FIGURE -5

On the third day of Siravyadham dialation as well as discolouration of veins has reduced to a very large extend.

classics, Varicose vein can be very much correlated to Siraja granthi. Being a kind of Raktadushti Vikara (Blood involved pathology), Raktamokshan and Rakta prasadana (blood purifying) internal medicines were given along with Vatanulomaka aushadhas (those which promote regular movement of Vata). As the case showed significant changes in the symptoms of pain, swelling, burning sensation, and dilated veins with discolouration, during the ten days of treatment with internal medicines as well as siravyadham done for 3 alternate days. It can be concluded that in such conditions of varicosed vein, Raktamokhsana along with Raktaprasadana drugs will be a choice of treatment. A large scale clinical study should be conducted to evaluate the efficacy and benefits of this modality of treatment with longer follow ups. ACKNOWLEDGEMENT:

CONCLUSION: Siravyadham is considered as half of entire treatment or even complete treatment in Shalya Tantra (surgery) because Rakta (blood) is considered as one among the major Adhistana (base) of vitiation. According to Ayurveda

We acknowledge Dr. Akhilesh Shukla, Assistant professor, Dept. of Samhita, Sanskrit and Siddhanta, Amrita School of Ayurveda, Kollam for the help during the preparation of the article as well as for the suggestions to improve the article.

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REFERENCES: Acharya Yadavji Trikamji (2009) Susrutha Samhita with Nibandhasangraha Commentary of Dalhanacharya and Nyayachandrika Panchika of GayadasaVaranasi, Chaukamba Orientalia, Nidana Sthana Chp: 11/8-9, Pp: 311. Sarira Sthana Chp: 8/22-23, Pp-383.

Acharya Parashuram Shastri Pt.Vidyasagar, (2006) Sarngadhara Samhita.Varanasi, Chaukamba Surbharati Prakashan, Pp. 373

Acharya Vaidya Harisastri Paradakara, (2011) Ashtanga Hridaya,with commentaries of Sarvanga Sundari of Arunadutta and Ayurveda Rasayana of Hemadri, 10th edn, Varanasi, Chikitsa Sthanam Chp:8/158, Pp-654., Chp:.22/35, and 22/41-44, Pp-731.

K.V Krishnan Vaidyan and S.Gopalapilla, (2006) Sahasrayogam, 26th edn, Malayalam, Vidyarambham publishers, kashaya prakarana Pp-40-291.

SOURCE OF SUPPORT:

NIL

Das S. (2014), A Concise textbook of Surgery, 8th edn, Kolkata: S. Das Chp-16 Pp256â&#x20AC;&#x201C;275.

Vaidya Harisastri Paradakara Acharya (2011) Ashtanga Hridaya, with commentaries of Sarvanga Sundari of Arunadutta and Ayurveda Rasayana of Hemadri, 10th edn, Varanasi. Uttara Sthana Chp: 30/7, Pp. 884.

CONFLICT OF INTEREST: None declared

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