Demand, End-Uses, and Conservation of Alpine Medicinal Plant Neopicrorhiza scrophulariiflora (Pennell) D. Y. Hong in Central Himalaya

Neopicrorhiza scrophulariiflora (Pennell) D. Y. Hong of Scrophulariaceae family (hereinafter referred to as Neopicrorhiza) has medicinally important rhizomes with high levels of trade. What factors drive demand for Neopicrorhiza in Central Himalaya is unknown. In this context, a nationwide comprehensive survey was conducted from September 2016 to March 2017 to assess demand, end-uses, and conservation of dry Neopicrorhiza rhizomes in Nepal. A total of 2313 herbal products were surveyed for Neopicrorhiza as an ingredient in 38 retailer shops. Processing industries of Neopicrorhiza in Nepal were interviewed using structured questionnaire. There were 23 herbal industries manufacturing 45 types of ayurvedic medicines as end-products containing Neopicrorhiza. The volume and value of annual demand for dry rhizomes of Neopicrorhiza in Nepal were found as 6076 kg and NRs 8573236 (USD 83235.30), respectively, in 2015/016 with average 264.17 kg/industry and NRs 1410.87 (USD 13.69) per kg. The major uses of ayurvedic medicines containing Neopicrorhiza were to treat a number of disease categories: cardiovascular system/liver (17), cardiovascular system/blood (6), nervous system (6), dermatological system (4), musculoskeletal system (3), digestive system (2), respiratory system (2), genitourinal system (4), and others (1). Despite changing legal regulation, trade and consumption of Neopicrorhiza exist in Nepal. It can be concluded that domestic consumption is not the major cause of resource depletion of Neopicrorhiza in Nepal.


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Evidence-Based Complementary and Alternative Medicine Source: Field survey (2015/16). * Herbal product categories mentioned in Table 1 were adopted from Smith-Hall et al. [28]. [23], phenylethanoid glycosides [24], cucurbitacin glycoside [25,26], and phenylpropanoid glycosides [27] having a wide range of biological activities. The dry and unprocessed rhizomes of Neopicrorhiza are under commercial trade. Though Nepal supplied 66 ± 12% of rhizomes of Neopicrorhiza in the global supply chain [30], indicating higher levels of harvesting, the domestic demand for this species in Nepal is unknown. Shrestha and Jha [6] reported that populations of Neopicrorhiza are fragmented and its population sizes appear to be declining despite no commercial collection at study period in trans-Himalayan dry valley of central Nepal, Manang district. Shrestha and Shrestha [31] reported unsustainable harvesting method of Neopicrorhiza in Rasuwa district of northern Nepal, where the plant was entirely uprooted with no part left for future regeneration. Ghimire et al. [5] reported that the size class of rhizomes of Neopicrorhiza harvested for trade was significantly smaller than those collected for health care, leading to premature harvesting. Sustainable harvesting of underground parts (rhizomes, roots, bulbs, or other storage organs) of long-lived species presents a particularly great challenge [32]. The high levels of trade and unsustainable harvesting can lead to pressure on the growing stock of Neopicrorhiza in Nepal, though currently national population status of this species is lacking. A rising demand for Himalayan plant based medicinal and cosmetic products was mentioned by Holley and Cherla [33] and Olsen [34] which could foster demand for raw materials, which hence could lead to rise in harvest level of medicinal and aromatic plant species. So it has become important to understand what is driving the demand for Neopicrorhiza in Nepal. Understanding end-uses of plant species is essential for prediction of future demand for the species and planning for its harvesting sustainability. However, nothing is known about the industrial consumption and end-products of Neopicrorhiza. The aim of this paper is to contribute to the understanding of the end-uses of commercially important medicinal plants by exploring the industrial demand for rhizomes of Neopicrorhiza and its endproducts, uses, and conservation in Nepal.

Materials and Methods
Step 1. Telephone survey: First of all, a list of herbal manufacturing industries in Nepal was compiled during September-November 2016 using a number of tools-web survey, online database analysis of Department of Plant Resources of Government of Nepal, database analysis of Department of Industry of Government of Nepal, analysis of directory of national census of manufacturing establishments (2069/2011), directories of herbal associations in Nepal, and stakeholder consultation. The output was a list of 246 herbal manufacturing industries.
Telephone interview adapted from Stanton and Futrell [35], Cooper and Schindler [36], and Boyd et al. [37] was conducted with representative of herbal manufacturing industries in November 2016 to find out the industries that process Neopicrorhiza. The main tool for the data collection with "telephone survey" was a structured questionnaire, finalized by expert panel and pretesting, and was composed of two major sections. Section 1 included introduction to the topic and researcher. Section 2 included categories of end-products the industry is currently producing, whether the industry processes Neopicrorhiza or not, name, location, and contact details of the industry.
Step 2: Market survey: The herbal products for Neopicrorhiza as an ingredient were randomly surveyed at 2313 herbal products at retailer shops in two big cities of Nepal, during November-December 2016 (see Table 1). We surveyed 1286 herbal products in Kathmandu city in 30 shops and 1027 herbal products at 8 retailer shops of Bharatpur city of Chitwan district of Nepal.
The main tool for the data collection in "market survey" was a structured questionnaire, finalized by expert panel and pretesting, and was composed of introduction, name of end-product containing Neopicrorhiza, form of product, manufacturers' details, who mainly buys, basis of sale (prescription or without prescription), use of product, and retail price.   (2), the permanent names of the provinces will be determined by a 2/3 vote of the respective province's legislature. They were officially indicated as 1,2,3,4,5, and 7 provinces. Province number 6 became the first province to get a name, as a provincial assembly meeting renamed it "Karnali" on 24 February 2018.
Step 3. Industrial survey: A total of 23 industries processing Neopicrorhiza were identified in Nepal and visited during January-March 2017, and face-to-face interviews were conducted with representatives of these industries using structured questionnaire, finalized by expert panel and pretesting.
The structured questionnaire was composed of three major sections. Section 1 included characteristics of respondents such as name, age, education, and number of years of experiences in processing of medicinal plants. Section 2 included characteristics of the industry such as name, year of establishment, and number of regular staff. Section 3 included name of end-product or end-product containing Neopicrorhiza, amount of its raw rhizomes needed, price paid for its raw rhizomes, and marketing channel of end-products. The information on end-products collected from market survey was also cross-checked and validated with this survey.
Respondents were informed about the purpose of the research and their prior consent was obtained for interview. The research did not require approval of ethics from government of Nepal since it was a pure academic research mainly based on noninvasive and nondestructive methods (i.e., interviews).
The standard terminology for categories of diseases was adopted from Kunwar et al. [29]. Data analysis was performed using MS Excel and IBM Statistics SPSS 20. Descriptive statistics including frequency, percentage, mean, sum, and average were used. The information was presented in tables and explained.

Characteristics of Processing Industries of Neopicrorhiza in Nepal.
Out of 246 herbal manufacturing industries, a total of 23 industries were found processing dry rhizomes of Neopicrorhiza in Nepal to manufacture its end-products. The number of Neopicrorhiza-processing industries in province three was the highest number of industries (n=13, 56.52%), while province four had the lowest number of those industries (n=1, 4.35%). The number of Neopicrorhiza-processing industries in provinces two and five was three (13.04%) and six (26.09%), respectively. There was not any Neopicrorhizaprocessing industries in province number 1, Karnali province, and province number 7 ( Figure 1). Out of the 23 Neopicrorhiza-processing industries recorded, 8 (34.78%) were located in Kathmandu district, 6 (26.09%) in Rupandehi district, and two (8.69%) in each of Parsa, Lalitpur (8.69%), and Makawanpur districts, and one (4.35%) was located in each of Chitwan, Gorkha, and Dhading districts. Herbal processing and manufacturing are still in their infancy in Nepal, though herbal industries have a great scope of providing employment and developing new natural products that can fetch better prices in international markets [38]. Winrock International et al. [39] identified only 15 major companies involved in the processing of 205 non-timber forest products for making ayurvedic preparations and essential oils. Edwards [40] had projected that Nepal will find it difficult to compete with India's highly complex, professionally managed herbal processing operations, which are on a considerably larger scale than may ever be developed in Nepal. We argue that there is a definite need for nation's priority towards the expansion of ayurvedic medicine and herbal manufacturing industries and to create conducive environment for competitive functioning in Nepal.
Characteristics of the industries that involved processing rhizomes of Neopicrorhiza are summarized in Table 2. All industries were established between the periods of 1963 and 2013. Except Singha Darbar Vaidhya Khana, all were privately owned (n=22, 95.65%). Majority of the industries (n=20, 86.95%) were established after 1992. The industries had different number of regular staff ranging from 2 to 50. Most of the industries (n=11; 47.83%) had 2-11 regular staff. The scales of Neopicrorhiza-processing industries were of small (30.43%), medium (34.78%), and large (34.78%) as perceived and reported by the respondents themselves.
Our research shows that majority of the Neopicrorhizaprocessing industries of Nepal emerged after 1992 AD, privately owned except one government owned Singha Darbar Vaidhya Khana, started during Rana regime for royal palace. The Industrial Enterprises Act of 1974 and its frequent amendments shifted the government's emphasis on growth from the public to the private sector in Nepal [41]. Nepal entered into multiparty democratic system in 1990 after a nationwide successful revolution, which increased private sector development in the country, including industries. The Industrial Policy of Nepal came into effect only in 2010 that identified priority industries for Nepal, including forest based industries, and ayurvedic and homeopathic medicine manufacturing, as priority industries [42].

Domestic Industrial Consumption (Volume and Value) of Neopicrorhiza in Nepal. The domestic industrial demand for
Neopicrorhiza in Nepal was unknown for a long time. Industrial survey shows that the annual demand for dry rhizomes of Neopicrorhiza in Neopicrorhiza-processing industries (23) in Nepal was 6076 kg (6.076 tons). It is novel finding. The demand for its rhizomes ranged from 5 to 1000 kg/yr in different industries, with average of 264.17 kg/yr per industry. We compared our findings with that of India where the annual domestic consumption of Picrorhiza kurroa was 415 MT [43]. Rhizomes of Picrorhiza kurroa (found in western Himalaya) and Neopicrorhiza scrophulariiflora (found in eastern and central Himalaya) are morphologically similar and enter into trade without differentiation. The annual legal trade of Neopicrorhiza in Nepal in fiscal year 2015/016 was 61 tons [44] which is around ten times higher than the domestic industrial demand for this species (6 tons) in 2015/016 as found in this study. Olsen [34] indicated with no doubt that the annual harvested amounts far exceed the domestic demand for medicinal and aromatic plants in Nepal, and we also found similar results for Neopicrorhiza. Olsen [30] estimated 100-400 tons of annual Neopicrorhiza export from Nepal. Though the export figures vary, these data indicate higher levels of export of dry Neopicrorhiza rhizomes from Nepal than domestic consumption. Province-wise annual demand for Neopicrorhiza in industries of Nepal in 2015/016 is presented in Table 3, with highest demand in province three and lowest in province four. None of the herbal industries located in provinces 1, 6 (hereinafter referred as Karnali), and 7 were found using Neopicrorhiza as raw material.
The industries paid NRs 1150 to 1600 (USD 11. 16 [30] reported that local traders paid NRs 87.9 (USD 0.853) in average to purchase one kilogram of dry rhizomes of Neopicrorhiza in 1997/98. Total annual value of dry rhizomes of Neopicrorhiza purchased for domestic industrial consumption in Nepal in 2015/016 was found NRs 8573236 (USD 83235.30) for 6.076 tons, which is much lesser than Olsen's estimate of 1.5 million USD annual export value of dry rhizomes of Neopicrorhiza from Nepal in 1997/98.

Availability of Neopicrorhiza for Industries and Its Substitutes.
All the respondents reported that their industries had not faced any difficulty in obtaining quality rhizomes of Neopicrorhiza to fulfill their annual demand. In most of the cases (18, 78.26%), the traders themselves made contact with the industries and provided sample of dry rhizomes of Neopicrorhiza. Then the specialists in industries inspected the quality of the rhizomes and made decision whether to purchase or not. They did not depend on single trader for purchase of the dry rhizomes of Neopicrorhiza. Only 6 (21.74%) industries contacted traders from their list to purchase the dry rhizomes of Neopicrorhiza. In both cases, price of the rhizomes was also the major competitive factor while deciding the purchase from particular trader. All the Neopicrorhiza-processing industries did not use any substitute for Neopicrorhiza, indicating that their demand for its rhizomes were easily fulfilled, they did not require large quantities of Neopicrorhiza annually, and Neopicrorhiza was only used as an ingredient in end-products. Plants gathered in Nepal are used either for domestic consumption or for sale and manufacturing the end-products in Nepal, India, and third countries [46], as also found in this study. The lower levels of demand for Neopicrorhiza and government's lifting of ban for collection and domestic trade might be the main reasons for ease in obtaining quality rhizomes of Neopicrorhiza by the industries.
Picrorhiza kurroa, of which India is a main consumer with annual domestic consumption of 415 MT [43], is open for trade in India. However, Olsen [30] argued that the majority of trade occurs in Neopicrorhiza from Nepal rather than Picrorhiza kurroa in India. Demand for Picrorhiza kurroa is continuously increasing in India; e.g., an annual growth rate of 12.9% was recorded for its demand for 220 tons in 2001-2002 and 317 tons during 2004-2005 [47]. China is apparently a major consumer of Neopicrorhiza [34]. This scenario indicates that there will definitely be pressure on supply status of Neopicrorhiza in future to industries due to this increasing regional demand. Multiple uses exert higher demand, leading to increased harvest, and such actions raise threats for medicinal plant species [18]. Regional level strategies will be necessary to keep balance between trade and conservation of Neopicrorhiza. Till date, population status of Neopicrorhiza in Nepal is unknown, without which any assumption of sustainable supply and trade of this species on long run might be misleading.

End-Products of Neopicrorhiza.
We did not know before what end-products were manufactured from Neopicrorhiza. Market and industrial surveys revealed that Neopicrorhiza was used for manufacturing forty-five (45) herbal products in Nepal. A complete list of those end-products with description is presented in Table 4. All those end-products were ayurvedic medicines. Neopicrorhiza was used only as an ingredient in these products. The end use of Neopicrorhiza for production of only ayurvedic medicines can be explained by the presence of medicinally useful iridoid glycosides such as picrosides I and II, and kutkoside in its rhizomes [48][49][50][51][52]. Out of 45 ayurvedic medicines containing Neopicrorhiza, 15 (33.33%) were classical (Classical Ayurvedic Medicine (CAM) is prepared by considering standard formulations from traditional Ayurvedic text books like Charaka Samhita, Sushruta Samhita, etc. Those formulations remain the same for specific medicines irrespective of the manufacturers.) and 30 (66.67%) were proprietary (Proprietary Ayurvedic Medicine (PAM) is prepared by industries using own formulations.) medicines. The forms of these medicines were different: 6 (13.33%) were in tablet form, 18 (40%) were in syrup form, 15 (33.33%) were in powder form, 5 (11.11%) were in gel form, and 1 (2.22%) was in capsule form.
The annual consumption of dry rhizomes of Neopicrorhiza for production of Hepadex (a liver tonic) was recorded as the highest (1000 kg). The annual consumption of dry rhizomes of Neopicrorhiza for production of particular end-product containing Neopicrorhiza ranged from 2 to 1000 kg. The average annual consumption of dry rhizomes of Neopicrorhiza for production of each product is 135.02 kg. The top three products, for production of which more than 500 kg dry rhizomes of Neopicrorhiza were purchased in a year, were Hepadex (a liver tonic), Rohitkyadi Churna (a liver medicine), and Hepagard DS (a liver tonic).

Production Rate of End-Products of Neopicrorhiza Adopted by
Industries. The number of industries producing particular product and rank of production of each product is presented in Table 4. Rohitkyadi Churna, Neembadi Churna, and Arogyavardini Vati are the top three end-products containing Neopicrorhiza in terms of production by higher number of industries. Among these products, Rohitkyadi Churna enjoyed the highest production rate of 14.5%. Edwards [40] argued that the technology behind much processing of nontimber forest products was relatively straightforward and a wide range of ayurvedic preparations was already produced in Nepal, albeit on a small scale. Our findings on the data of annual demand for dry Neopicrorhiza rhizomes in domestic industries (6.076 tons) and their expense on these rhizomes (USD 83235.30) is consistent with Edwards's general view of small scale production of ayurvedic medicines within Nepal.
Nepali and Indian manufacturers of Ayurvedic medicines available in the Nepalese markets are nearly equal in number, but the Indian products are dominating the Nepalese market [53]. It is interesting to note that, out of 45 Neopicrorhiza-containing ayurvedic medicines, only three of those medicines were produced by more than one industry (Table 4). It shows that industries of Nepal have not yet diversified the Neopicrorhiza-containing ayurvedic medicines in Evidence-Based Complementary and Alternative Medicine 7 Table 4: End-products of Neopicrorhiza with their processing industries, consumption, uses, and production rank in 2015/16.   [29]) Note: The liver is included for cardiovascular disease category because liver filters or detoxifies all the blood coming from the digestive tract before the blood circulates back into the heart. In human body, liver is the only organ having two blood supplies: hepatic artery bringing blood from heart and hepatic portal vein brining blood from the intestines.

Rank of production
their production system. More processing of plants and the production of end-products in Nepal would greatly increase the value of the plants to the national economy [54]. Around only ten percent herbal industries out of 246 surveyed herbal industries in Nepal were found processing Neopicrorhiza rhizomes to produce the end-products. Hence, when issues emerge on resource depletion of Neopicrorhiza in Nepal, we here argue that it is due to export of Neopicrorhiza from Nepal than from domestic consumption, and regulatory measures should address the trade of Neopicrorhiza in terms of foreign export.

Uses of End-Products of Neopicrorhiza. We found that
Neopicrorhiza is used to produce only Ayurvedic medicines. The major uses of the ayurvedic medicines containing Neopicrorhiza were recorded to treat a number of illness categories: cardiovascular, nervous, dermatological, musculoskeletal, genitourinal, respiratory, digestive, and others. The highest number of products (n=17, 37.78) was reported to treat disease associated with liver. Previous studies had explored the use of the unprocessed rhizomes of Neopicrorhiza to treat various ailments such as liver disorders, fever, asthma, and jaundice and have pharmaceutical value for hepatoprotective, immunomodulator, and antiasthmatic activities [22,55,56] in Indian, Bhutanese, Tibetan, and Chinese traditional medicines. Mulliken and Crofton [57] argued that it is not inconceivable that in future the flow of Picrorhiza/Neopicrorhiza supplies could shift to the north (China), particularly if the efficacy of Picrorhiza kurroa in the treatment of liver disease is confirmed. The information about major use (indication) of the ayurvedic medicine provided in the label of the medicines was cross-checked and validated during market survey and industrial survey and was found consistent with information of Table 5.  (Table 6). In 2001, Government of Nepal banned for collection, utilization, sale, distribution, transport, and export of Neopicrorhiza [58]. In 2003, the government had revised the restriction based on Forest Act 1993 [59]. The government has set conditions for export of this species: (1) clear taxonomic identification of the Neopicrorhiza and its confirmation by the Department of Plant Resources are mandatory. (2) Department of Forest then permits the export after assessing the availability of the species.
Government of Nepal has changed the royalty rates of medicinal and aromatic plant products from time to time, including Neopicrorhiza [60]. In 2005, the royalty rate was NRs 10/kg [61].  (Table 7). 33.652 tons of Neopicrorhiza was collected and traded from Humla district during the fiscal year 1999-2000 AD (2055-056 BS) [62]. The government's official records show that the total volume of dry rhizomes of Neopicrorhiza exported to India, after receiving CITES certificate, from Nepal during 2015/016 period was around 28 tons [44]. Excessive and unregulated commercial harvest of medicinal plants (legally and illegally) has caused direct threat to the high value species [63]. Apart from unsustainable harvesting, other factors such as debris deposition at pastureland and cliff, deforestation, habitat encroachment, overgrazing, wildfires, shifting cultivation, and climate change contribute to species loss [4,17,18,29,64].
Looking at the province-wise trade, the legally traded volumes of Neopicrorhiza in provinces 3,4, Karnali, and 7 were 1.    [34]. Olsen [30] estimated 100-400 tons of annual Neopicrorhiza export from Nepal in 1998/99. These estimates seemed much higher when compared to legally traded volume of Neopicrorhiza in 2015/016 (i.e., 61 tons). This variation could hint for possibility of existence of illegal trade of Neopicrorhiza through Nepal. The main challenge will be how to regulate the export of this species outside the country while meeting the domestic demand and maintaining sustainable growing stock of this species. However, currently the domestic consumption of Neopicrorhiza cannot be entitled to be the root cause of resource depletion of this species.

Conclusions
This research presents a first exploration of domestic demand for alpine medicinal plant Neopicrorhiza, and its end-uses and manufacturers in Nepal. The annual domestic demand for dry rhizomes of Neopicrorhiza in Nepal was found around six tons in case year 2015/016 and this value is lesser than that of total traded volume and export. Forty-five end-products were found containing Neopicrorhiza as an ingredient and all those products were ayurvedic medicines. The major uses of the ayurvedic medicines containing Neopicrorhiza were to treat diseases associated with cardiovascular, nervous, dermatological, musculoskeletal, genitourinal, respiratory, and digestive systems. It can be concluded that the domestic consumption is not the major cause of resource depletion of Neopicrorhiza in Nepal. Regional consideration is necessary in setting priorities for conservation of Neopicrorhiza and sustainable trade in and from Nepal. In addition, growing stock of this species should be quantified throughout its habitat to set such priorities. Similar studies on end uses of Neopicrorhiza are recommended in countries (e.g., India) where this species is exported from Nepal.

Data Availability
The data used to support the findings of this study are available from the corresponding author upon written request. The questionnaire used in industrial survey is available in Smith-Hall et al. [28].