Antibacterial Efficacy of Green Synthesized Silver Nanoparticles Using Salvia nubicola Extract against Ralstonia solanacearum, the Causal Agent of Vascular Wilt of Tomato.

Ralstonia solanacearum is a phytopathogen causing bacterial wilt diseases of tomato and affecting its productivity, which leads to prominent economic losses annually. As an alternative to conventional pesticides, green synthesized nanoparticles are believed to possess strong antibacterial activities besides being cheap and ecofriendly. Here, we present the synthesis of silver nanoparticles (Sn-AgNPs) from medicinally important aqueous plant extracts of Salvia nubicola. Characterization of biologically synthesized nanoparticles was performed through UV-vis spectrophotometry, Fourier transform infrared spectroscopy (FTIR), energy-dispersive X-ray spectroscopy, X-ray diffraction, scanning electron microscopy, transmission electron microscopy (TEM), and thermogravimetric analysis. The antibacterial activity of the biosynthesized silver nanoparticles was tested against the phytopathogen R. solanacearum through in vitro experiments. Preliminary phytochemical analysis of the plant extracts revealed the presence of substantial amounts of flavonoids (57.08 mg GAE/g), phenolics (42.30 mg GAE/g), tannins, and terpenoids. The HPLC phenolic profile indicated the presence of 25 possible bioactive compounds. Results regarding green synthesized silver nanoparticles revealed the conformation of different functional groups through FTIR analysis, which could be responsible for the bioreduction and capping of Ag ions into silver NPs. TEM results revealed the spherical, crystalline shape of nanoparticles with the size in the range of 23-63 nm, which validates SEM results. Different concentrations of Sn-AgNPs (T1 (500 µg/mL) to T7 (78.1 µg/mL)) with a combination of plant extracts (PE-Sn-AgNPs) and plant extracts alone exhibited an efficient inhibition of R. solanacearum. These findings could be used as an effective alternative preparation against the bacterial wilt of tomato.

High throughput biochemical profiling, and functional potential analysis for valorization of grape peduncles.

Vitis vinifera L., commonly known as grape is a major fruit crop in the world. Grapes seem to confer health benefits due to their chemical components, biological and antioxidant activities. The present study is conducted to evaluate the biochemical constituents, antioxidant, and antimicrobial potential of ethanolic grape peduncles (EGP) extract. The result of phytochemical analysis revealed the presence of various phytochemicals such as flavonoid, tannin, carbohydrates, alkaloids, cardiac glycoside, phenol, steroid, terpenoids, quinones and anthraquinones. Furthermore, total phenolic content (TPC) and total flavonoid contents (TFC) were 7.35 ± 0.25 mg GAE/g (Gallic Acid Equivalent per gram) and 29.67 ± 0.13 mg QE/g (Quercetin Equivalent per gram) respectively. DPPH (2, 2-diphenyl-1-picrylhydrazyl) free radical scavenging assay revealed IC50 = 159.3 µg/mL. The antibacterial and antifungal study disclosed that the extract was highly potent against Salmonella typhi with maximum zone of inhibition of 27.2 ± 1.60 mm and Epidermophyton floccosum with 74 ± 1.81% inhibition. The extract was analyzed for its cytotoxicity and antileishmanial activity and showed no activity against HeLa cell line and promastigotes of Leishmania major. Elements Fe, Mn, Ni, Pb and Cd were determined by atomic absorption spectroscopy and approximately 50 compounds were identified by Gas Chromatography-Mass Spectrometry (GC-MS). Current work suggest that grape peduncles can be a promising source of bioactive medicinal component.

Traditional Uses of Wild Edible Mushrooms among the Local Communities of Swat, Pakistan.

Mushrooms play a crucial role in human life as well as in nature, providing food, medicine, and carrying out vital processes of decomposition, nutrient recycling, and developing mycorrhizal association with plants. The traditional system of knowledge about identification, collection, and usage of mushrooms has been accumulated through the shared experiences of many generations. Unfortunately, there have been continuous threats to the folk knowledge of mushrooms mainly due to habitat degradation, urbanization, and contemporary medication. The current research was, therefore, aimed to document an ethnomycological knowledge possessed by the ethnic communities of Swat, Pakistan. The purposive randomized sampling was carried out using chain referral method. Ethno-mycological information was collected from 62 informants using free listing, preference ranking, and use totaled methods. In total, 34 species of mushrooms belonging to 31 genera and 21 families were reported. About 85% of the reported species belong to Basidiomycetes, and 12.5% to Ascomycetes are used as food and for medicinal purposes. Morchella angusticeps, M. esculenta, Pleurotus sp., Auricularia sp., Flammulina velutipes, Agaricus bisporus, Ganoderma lucidum, and Sanghuangporus sanghuang were among the most cited edible and medicinal mushrooms. The current study revealed that district Swat is rich in wild edible and medicinal mushrooms (WEMs), and the local communities possess rich traditional knowledge about their collection, storage, and utilization. The diversity of WEMs of this region could contribute substantially to the socio-economic uplifting of the local communities through appropriate domestication and commercialization. Anthropogenic factors, coupled with depletion of traditional knowledge, threaten the diversity of WEMs in the region; therefore, in situ and ex situ conservation strategies are highly recommended.

A comprehensive review on traditional uses, phytochemistry and pharmacological properties of Paeonia emodi Wall. ex Royle: current landscape and future perspectives.

Paeonia emodi Wall. ex Royle is commonly known as Himalayan paeony has great importance as a food and medicine. The practice of Paeonia emodi Wall. ex Royle is very ancient and it is conventionally used for a wide range of illnesses in the folk system of medicine because of its wide beneficial phytochemical profile. The main purpose of the current review was the synthesis of recent data on botany, ethnopharmacology, phytochemistry and potential pharmacological mechanisms of action of Paeonia emodi Wall. ex Royle, thus offering new prospects for the development of new adjuvant natural therapies. Using scientific databases such as PubMed/MedLine, Scopus, Web of Science, ScienceDirect, Google Scholar, Springer, and Wiley, a comprehensive literature search was performed for Paeonia emodi Wall. ex Royle. For searching, we used the next MeSH terms: "Biological Product/isolation and purification", "Biological Products/pharmacology", "Drug Discovery/methods", "Ethnopharmacology, Medicine", "Traditional/methods", "Paeonia/chemistry", "Plant Extracts/pharmacology", "Phytochemicals/chemistry", "Phytochemicals/pharmacology", "Plants, Medicinal". The results of the most recent studies were analyzed and the most important data were summarized in tables and figures. Phytochemical research of Paeonia emodi Wall. ex Royle has led to the isolation of triterpenes, monoterpenes, phenolic acids, fatty acids, organic compounds, steroids, free radicals and some other classes of primary metabolites. In addition, diverse pharmacological activities like antibacterial, antifungal, anticoagulant, airway relaxant lipoxygenase and beta-glucuronidase inhibiting activity, radical scavenging activity, phytotoxic and insecticidal activities have been reported for Paeonia emodi Wall. ex Royle. Different bioactive compounds of Paeonia emodi Wall. ex Royle has proven their therapeutic potential in modern pharmacological and biomedical research to cure numerous gastrointestinal and nervous disorders. In future, further in vitro and in vivo therapeutic studies are required to identify new mechanisms of action, pharmacokinetics studies, and new pharmaceutical formulations for target transport and possible interaction with allopathic drugs. Also, new research regarding quality evaluation, toxicity and safety data in humans is needed.

A Novel Based Synthesis of Silver/Silver Chloride Nanoparticles from Stachys emodi Efficiently Controls Erwinia carotovora, the Causal Agent of Blackleg and Soft Rot of Potato.

In recent years, the biological synthesis of silver nanoparticles has captured researchers' attention due to their unique chemical, physical and biological properties. In this study, we report an efficient, nonhazardous, and eco-friendly method for the production of antibacterial silver/silver chloride nanoparticles utilizing the leaf extract of Stachys emodi. The synthesis of se-Ag/AgClNPs was confirmed using UV-visible spectroscopy, DPPH free radical scavenging activity, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and X-ray diffraction (XRD). An intense peak absorbance was observed at 437 nm from the UV-visible analysis. The Stachys emodi extract showed the highest DPPH scavenging activity (89.4%). FTIR analysis detected various bands that indicated the presence of important functional groups. The SEM morphological study revealed spherical-shaped nanoparticles having a size ranging from 20 to 70 nm. The XRD pattern showed the formation of a spherical crystal of NPs. The antibacterial activity performed against Erwinia carotovora showed the maximum inhibition by centrifuged silver nanoparticles alone (se-Ag/AgClNPs) and in combination with leaf extract (se-Ag/AgClNPs + LE) and leaf extract (LE) of 98%, 93%, and 62% respectively. These findings suggested that biosynthesized NPs can be used to control plant pathogens effectively.

Occurrence and Distribution of Heavy Metals in Mining Degraded Soil and Medicinal Plants: A Case Study of Pb/Zn Sulfide Terrain Northern Areas, Pakistan.

Mining activities have serious environmental impacts, thus releasing heavy metals (HMs) such as cadmium (Cd), lead (Pb), chromium (Cr), zinc (Zn) and nickel (Ni) into the surrounding environment. The current paper investigated the impacts of mining activities of Pb-Zn sulfide on soil and medicinal plants. Hence, soil samples (n = 36) and medicinal plants (n = 36) samples were collected, acid extracted and were analyzed through Inductively Coupled Plasma Mass Spectrometry (ICP-MS) for HMs quantification. Our results showed that mineralized zones showed high HMs enrichment levels as compared to non-mineralized zones. Various Indices for HMs assessment revealed that the contaminated soil of the study area had low to extreme level. The mean concentrations of HMs in mining degraded soil and medicinal plants were significantly higher (p ≤ 0.01) and were found in order of Zn > Pb > Cr > Ni > Cd and Cr > Zn > Pb > Ni > Cd respectively. Similarly, some widely consumable medicinal plants showed good metal accumulation for Cd, Cr and Pb i.e., 3.57 mg kg1, 350 mg kg-1 and 335 mg kg-1 in C. sativa, while 5.9 mg kg-1, 276.9 mg kg-1 and 188.7 mg kg-1 in R. hestatus respectively. Hence, the present study recommended that medicinal plants grown in mining areas should be analyzed for HMs concentration before consumption.

Biosynthesized silver nanoparticles using Polygonatum geminiflorum efficiently control fusarium wilt disease of tomato.

Nanomaterials are gaining tremendous potential as emerging antimicrobials in the quest to find resistance-free alternatives of chemical pesticides. In this study, stable silver nanoparticles were synthesized using the aqueous extract of medicinal plant species Polygonatum geminiflorum , and their morphological features were evaluated by transmission electron microscopy, X-ray diffraction spectroscopy and energy dispersive X-ray analysis. In vitro Antifungal activity of the synthesized silver nanoparticles (AgNPs) and P. geminiflorum extract (PE) either alone or in combination (PE-AgNPs) against Fusarium oxysporum was evaluated using disc-diffusion and well-diffusion methods. In planta assay of the same treatments against Fusarium wilt diseases of tomato was evaluated by foliar spray method. Moreover, plant extract was evaluated for the quantitative investigation of antioxidant activity, phenolics and flavonoids by spectroscopic and HPLC techniques. Phytochemical analysis indicated the presence of total phenolic and flavonoid contents as 48.32 mg ± 1.54 mg GAE/g and 57.08 mg ± 1.36 mg QE/g, respectively. The DPPH radical scavenging of leaf extract was found to be 88.23% ± 0.87%. Besides, the HPLC phenolic profile showed the presence of 15 bioactive phenolic compounds. Characterization of nanoparticles revealed the size ranging from 8 nm to 34 nm with average crystallite size of 27 nm. The FTIR analysis revealed important functional groups that were responsible for the reduction and stabilization of AgNPs. In the in vitro assays, 100 µg/ml of AgNPs and AgNPs-PE strongly inhibited Fusarium oxysporum. The same treatments tested against Fusarium sprayed on tomato plants in controlled environment exhibited nearly 100% plant survival with no observable phytotoxicity. These finding provide a simple baseline to control Fusarium wilt using silver nano bio-control agents without affecting the crop health.

Green Synthesis of Silver Nanoparticles Using Euphorbia wallichii Leaf Extract: Its Antibacterial Action against Citrus Canker Causal Agent and Antioxidant Potential.

Biologically synthesized silver nanoparticles are emerging as attractive alternatives to chemical pesticides due to the ease of their synthesis, safety and antimicrobial activities in lower possible concentrations. In the present study, we have synthesized silver nanoparticles (AgNPs) using the aqueous extract of the medicinal plant Euphorbia wallichii and tested them against the plant pathogenic bacterium Xanthomonas axonopodis, the causative agent of citrus canker, via an in vitro experiment. The synthesized silver nanoparticles were characterized by techniques such as UV-Vis spectroscopy, Fourier transform infrared spectroscopy, energy-dispersive X-ray spectroscopy, X-ray diffraction analysis and transmission electron microscopy. Moreover, the plant species were investigated for phenolics, flavonoids and antioxidant activity. The antioxidant potential of the extract was determined against a DPPH radical. The extract was also evaluated for phenolic compounds using the HPLC technique. The results confirmed the synthesis of centered cubic, spherical-shaped and crystalline nanoparticles by employing standard characterization techniques. A qualitative and quantitative phytochemical analysis revealed the presence of phenolics (41.52 mg GAE/g), flavonoids (14.2 mg QE/g) and other metabolites of medicinal importance. Different concentrations (1000 µg/mL to 15.62 µg/mL-2 fold dilutions) of AgNPs and plant extract (PE) alone, and both in combination (AgNPs-PE), exhibited a differential inhibition of X. axanopodis in a high throughput antibacterial assay. Overall, AgNPs-PE was superior in terms of displaying significant antibacterial activity, followed by AgNPs alone. An appreciable antioxidant potential was recorded as well. The observed antibacterial and antioxidant potential may be attributed to eight phenolic compounds identified in the extract. The Euphorbia wallichii leaf-extract-induced synthesized AgNPs exhibited strong antibacterial activity against X. axanopodis, which could be exploited as effective alternative preparations against citrus canker in planta in a controlled environment. In addition, as a good source of phenolic compounds, the plant could be further exploited for potent antioxidants.

Antioxidant, Anticancer, and PXR-Dependent CYP3A4 Attributes of Schweinfurthia papilionacea (Burm.f.) Boiss., Tricholepis glaberrima DC. and Viola stocksii Boiss.

Present study established the biological potential of Schweinfurthia papilionacea, Tricholepis glaberrima and Viola stocksii extracts for their potential applications in drug formulations. Initially, FTIR was performed to ascertain functional groups and then plant extracts were prepared using five solvents depending on the polarity. Total phenolic contents were observed in the range of 36.36 ± 1.08 mg GAE/g to 95.55 ± 2.46 mg GAE/g while flavonoid contents were found in the range of 10.51 ± 0.25 mg QE/g to 22.17 ± 1.79 mg QE/g. Antioxidant activity was determined using TRP, CUPRAC, TAC and DPPH assays and was recorded highest in S. papilionacea followed by T. glaberrima extracts. TPC and TFC were found to be strongly correlated with TRP (r > 0.50), CUPRAC (r > 0.53) and DPPH (r = 0.31 and 0.72) assay while weakly correlated with TAC (r = 0.08 and 0.03) as determined by Pearson correlation analysis. Anticancer activity showed that S. papilionacea chloroform extracts possess highest cell viability (85.04 ± 4.24%) against HepG2 cell lines while T. glaberrima chloroform extracts exhibited highest activity (82.80 ± 2.68%) against HT144 cell lines. Afterwards, highest PXR activation was observed in T. glaberrima (3.49 ± 0.34 µg/mL fold) at 60 µg/mL and was correlated with increase in CYP3A4 activity (15.0 ± 3.00 µg/mL IC50 value). Furthermore, antimalarial activity revealed >47600 IC50 value against P. falciparum D6 and P. falciparum W2 and antimicrobial assay indicated highest activity (32 ± 2.80 mm) in S. papilionacea against C. neoformans. At the end, GC-MS analysis of n-hexane plant extracts showed 99.104% of total identified compounds in T. glaberrima and 94.31% in V. stocksii. In conclusion, present study provides insight about the different biological potentials of S. papilionacea and T. glaberrima extracts that rationalize the applications of these extracts in functional foods and herbal drugs for the management of oxidative-stress related diseases, antimicrobial infections and liver and skin cancer.

Biogenic synthesis of iron oxide nanoparticles via Skimmia laureola and their antibacterial efficacy against bacterial wilt pathogen Ralstonia solanacearum.

Plant diseases are threat to global food security. The excessive use of agrochemicals is the leading cause of pesticides resistance and toxicity to beneficial life forms. The quest for innocuous and alternate antimicrobial agent is crucial in order to overcome the pathogen resistance and the birth of nanotech offers pledge to combat pathogenic organisms. In this study, a facile benign biogenic approach was adopted for the synthesis of biocompatible iron oxide nanoparticles (Fe2O3-NPs) via Skimmia laureola leaf extract and the synthesized nanoparticles were evaluated for their antibacterial efficacy against bacterial wilt pathogen Ralstonia solanacearum in vitro and in planta. Physico-chemical characterization of the synthesized nanoparticles was performed through UV-Visible spectroscopy, Fourier Transform Infrared Spectroscopy, X-Ray Diffraction, Energy Dispersive X-ray Spectroscopy and Scanning Electron Microscopy. The results revealed polydisperse nanoparticles in the size range of 56 nm to 350 nm. The culture media containing 6 mg/mL of Fe2O3-NPs dramatically inhibited the bacterial growth in vitro. Scanning electron microscopy revealed degenerative characteristics including degraded, shriveled and concentrated cell walls. Diseases severity was effectively reduced with 6% w/v of Fe2O3-NPs treated root zone in planta. Plant shoots, root length and fresh biomass were enhanced with Fe2O3-NPs treatments. The results indicated that the biosynthesized Fe2O3-NPs have the potential to control agriculturally important phytopathogen Ralstonia solanacearum in vitro and in planta.

Traditional use of medicinal plants among Kalasha, Ismaeli and Sunni groups in Chitral District, Khyber Pakhtunkhwa province, Pakistan.

ETHNOPHARMACOLOGICAL RELEVANCE: The traditional use of medicinal plants for the treatment of human and livestock ailments is important to indigenous communities in the northern parts of Pakistan, and considered to be a valuable local biological and sociocultural heritage. The aim of this study was to obtain a detailed inventory of medicinal plant use and preparation among Kalasha, Ismaeli and Sunni groups. MATERIALS AND METHODS: Semi-structured group and individual interviews were carried out with men and women of different age groups that identified themselves as being Kalasha, Ismaeli or Sunni. Interviews were followed up by field visits to collect herbarium vouchers and record in greater detail the exact methods of harvesting, preparation and use on medicinal plants. RESULTS: A total of 76 species were recorded for treatment of various diseases. The Kalasha, Ismaili and Sunni ethnic groups have similar medicinal floras, but show striking differences in plant use. Our comparative survey shows that out of all species reported in this study, only 13 species have been reported previously from Chitral District. CONCLUSIONS: Indigenous knowledge of folk medicine is intricately linked to local culture, religion and history. Any short study can only scratch the surface of this intricate system, but provide an insight into the critical importance of medicinal plants for local livelihoods and the important role these play in health care systems. There is a great need to assess and properly manage the production potential of medicinal plants to ensure sustainable supply of these species for local use and subsistence trade.

Indigenous knowledge of folk medicines among tribal minorities in Khyber Pakhtunkhwa, northwestern Pakistan.

ETHNOPHARMACOLOGICAL RELEVANCE: Mapping ethnomedicinal plants and associated indigenous knowledge of folk medicines can provide a comprehensive overview of individual herbs employed in health care. Reliance on medicinal plants in remote parts of northern Pakistan is high, especially among women, but no research has investigated specifically which plants are used. This study investigated indigenous knowledge of folk medicines among tribal minorities in selected sites in upper Swat, Buner and Chitral Districts in Khyber Pakhtunkhwa Province. MATERIALS AND METHODS: Interviews were conducted with gender-specific focus groups using questionnaires and standardized data sheets, followed by forest walks in each of the visited areas. General medicinal herb use, preparations, storage, marketing and collection habits for each gender group were ascertained from the questionnaires. RESULTS: In total 168 women and 390 men were interviewed and provided information on 127 different shared medicinal species. Species use consensus among the informants ranged from 2.3% to 83.3%, with Cynodon dactylon, Avena sativa, Celtis australis, Datura stramonium, Solanum nigrum, Skimmia laureola, Spiraea nervosa, Ziziphus jujuba, Rumex hastatus, Plantago lanceolata, Lathyrus aphaca and Ficus palmata having the highest reported consensus. The survey also revealed that a number of medicinal species were exploited by the community for both marketing and personal use, and many of these species were reported as being rare, vulnerable or even endangered. CONCLUSIONS: The results revealed that women in all the three districts were important custodians of medicinal plant knowledge, but elder women in general and the women from Buner district in particular had a superior understanding of folk medicine. The forest walks revealed that women׳s traditional medicinal knowledge was based on a more limited diversity of plant species. People in tribal communities have an expressed interest in learning efficient techniques for medicinal plant collection, preparation, storage and cultivation advice, and to learn more about the potential of marketing medicinal herbs and ways to reach local market centers. Education and awareness were considered to be essential for improved health care and successful marketing.

Economic benefits of high value medicinal plants to Pakistani communities: an analysis of current practice and potential.

BACKGROUND: Poverty is pervasive in the Swat Valley, Pakistan. Most of the people survive by farming small landholdings. Many earn additional income by collecting and selling plant material for use in herbal medicine. This material is collected from wild populations but the people involved have little appreciation of the potential value of the plant material they collect and the long term impact their collecting has on local plant populations. METHODS: In 2012, existing practices in collecting and trading high value minor crops from Swat District, Pakistan, were analyzed. The focus of the study was on the collection pattern of medicinal plants as an economic activity within Swat District and the likely destinations of these products in national or international markets. Local collectors/farmers and dealers were surveyed about their collection efforts, quantities collected, prices received, and resulting incomes. Herbal markets in major cities of Pakistan were surveyed for current market trends, domestic sources of supply, imports and exports of herbal material, price patterns, and market product-quality requirements. RESULTS: It was observed that wild collection is almost the only source of medicinal plant raw material in the country, with virtually no cultivation. Gathering is mostly done by women and children of nomadic Middle Hill tribes who earn supplementary income through this activity, with the plants then brought into the market by collectors who are usually local farmers. The individuals involved in gathering and collecting are largely untrained regarding the pre-harvest and post-harvest treatment of collected material. Most of the collected material is sold to local middlemen. After that, the trade pattern is complex and heterogeneous, involving many players. CONCLUSIONS: Pakistan exports of high value plants generate over US$10.5 million annually in 2012, with a substantial percentage of the supply coming from Swat District, but its market share has been declining. Reasons for the decline were identified as unreliable and often poor quality of the material supplied, length of the supply chain, and poor marketing strategies. These problems can be addressed by improving the knowledge of those at the start of the supply chain, improving linkages among all steps in the chain, and developing sustainable harvesting practices.

Economically and ecologically important plant communities in high altitude coniferous forest of Malam Jabba, Swat, Pakistan.

A study on the economically important plant communities was carried out during summer 2008 in various parts of Malam Jabba valley, Swat. The principal aim of the study was phytosociological evaluation with special reference to the occurrence of commercially important medicinal plant species in coniferous forest of the study area. Secondly to prepare ethnobotanical inventory of the plant resources of the area, as well as to evaluate the conservation status of important medicinal and aromatic plants (MAPs) through rapid vulnerable assessment (RVA) procedure. The study documented 90 species of ethnobotanical importance, out of these 71 spp used as medicinal plant, 20 spp fodder plant, 10 spp vegetables, 14 spp wild fruit, 18 spp fuel wood, 9 spp furniture and agricultural tools, 9 spp thatching, fencing and hedges, 4 spp honey bee, 2 spp evil eyes, 2 spp religious and 3 spp as poison. Phytosociologically six plant communities were found, comprising five herbs-shrubs-trees communities and one meadow community. Further study is, therefore, required to quantify the availability of species and to suggest suitable method for their production and conservation. Recommendations are also given in the spheres of training in identification, sustainable collection, value addition, trade monitoring and cooperative system of marketing.

Assessment of local management practices on the population ecology of some medicinal plants in the coniferous forest of Northern Parts of Pakistan.

A study on the assessment of local management practices on the population of three medicinal plants viz.: Persicaria amplexicaule. D. Don., Valeriana jatamansi Jones and Viola serpens Wall ex Roxb was conducted during 2002-2004 in the coniferous forest of Northern Parts of Pakistan. The objective of the study was to know the impact of current management practices on the population size of targeted plants. The study showed that the involvement of locals in the gathering of targeted plants varied with the change in elevation. Among the targeted plants V. serpens was collected by large majorities of people (83.3%) at 2700 m followed by 72% at 2300 m and 37% at 1900 m. V. jatamansi was harvested by a small number of people (18.1%) at 1900 and 2300 m each, followed by 8.3% at 2700 m. While P. amplexicaule was harvested by a few collectors (9.1%) at 1900 m and 9.6% at 2300 m followed by 8.3% at 2700 m. The study concluded that these species have been extracted so heavily in the past that they are found now sparsely in some sites of the study area. Secondly, due to loss of its habitat by deforestation and encroachment of land for cultivation its population is on the decline towards extinction. Therefore, the current study recommends the conservation of the remaining populations of targeted plants through active participation of local communities.