Background: Wild edible plants (WEP) provide staple and supplement foods, as well as cash income to local communities, thus favouring food security. However, WEP are largely ignored in land use planning and implementation, economic development, and biodiversity conservation. Moreover, WEP-related traditional knowledge is rapidly eroding. Therefore, we designed this study to fulfill a part of the knowledge gap by providing data on diversity, traditional knowledge, economic potential, and conservation value of WEP from Nepal. Methods: The information was collected through focus group discussions and key informant interviews. Percentage of general utility of the plants among the study communities was evaluated using the Chi-square (χ²) test of homogeneity. High priority species were identified after consultation with the local stakeholders followed by scoring based on defined criteria. Pairwise ranking was used to assess ethnoecological knowledge to identify the threats to WEP. Results: We documented 81 species belonging to Angiosperms (74), Pteridophytes (5), and Fungi (2). Most of the species were used as fruits (44 species) followed by vegetables (36). Almost half of the species (47%) were also used for purposes other than food. From the species with market value (37% of the total), 10 were identified as high priority species. Pairwise ranking revealed that WEP are threatened mostly by habitat destruction, land-use change and over-harvesting. Some of these plants are crop wild relatives and could thus be used for crop improvement. Interestingly, our study also revealed that young people who spend most of the time in the forest as herdsmen are particularly knowledgeable of wild fruit plants. Conclusion: We provide empirical evidence from a relatively large area of Nepal about diversity and status of WEP, as well as methodological insights about the proper knowledge holders to consult. Regarding the unique and important knowledge they have on WEP, young people should be included when recruiting participants to ethnobotanical studies or to any type of consultation about WEP. The habit of using wild edible plants is still alive and is a traditional culinary practice that demonstrates rich traditional knowledge of local people. WEP were found to be important for livelihood as well as showing great potential for crop improvement. Priority species should be promoted for income generation activities through sustainable collection and trade. Communities should engage in minimizing the threats to these valuable resources.

Non-timber Forest Products (NTFPs), an important provisioning ecosystem services, are recognized for their contribution in rural livelihoods and forest conservation. Effective management through sustainable harvesting and market driven commercialization are two contrasting aspects that are bringing challenges in development of NTFPs sector. Identifying potential species having market value, conducting value chain analyses, and sustainable management of NTFPs need analysis of their use patterns by communities and trends at a regional scale. We analyzed use patterns, trends, and challenges in traditional use and management of NTFPs in the southern slope of Kangchenjunga Landscape, Eastern Himalaya and discussed potential implications for conservation and livelihoods. A total of 739 species of NTFPs used by the local people of Kangchenjunga Landscape were reported in the reviewed literature. Of these, the highest number of NTFPs was documented from India (377 species), followed by Nepal (363) and Bhutan (245). Though the reported species were used for 24 different purposes, medicinal and edible plants were the most frequently used NTFP categories in the landscape. Medicinal plants were used in 27 major ailment categories, with the highest number of species being used for gastro-intestinal disorders. Though the Kangchenjunga Landscape harbors many potential NTFPs, trade of NTFPs was found to be nominal indicating lack of commercialization due to limited market information. We found that the unsustainable harvesting and lack of marketing were the major constraints for sustainable management of NTFPs sector in the landscape despite of promising policy provisions. We suggest sustainable harvesting practices, value addition at local level, and marketing for promotion of NTFPs in the Kangchenjunga Landscape for income generation and livelihood improvement that subsequently contributes to conservation.

This study aimed to isolate, purify, and characterize a potential thermophilic cellulase-producing bacterium from the Himalayan soil. Eleven thermophilic bacteria were isolated, and the strain PANG L was found to be the most potent cellulolytic producer. Morphological, physiological, biochemical, and molecular characterization identified PANG L as Bacillus licheniformis. This is the first study on the isolation of thermostable cellulase-producing Bacillus licheniformis from the Himalayan soil. This bacterium was processed for the production of cellulase enzyme. The optimum conditions for cellulase production were achieved at 45C after 48h of incubation at pH 6.5 in media-containing carboxymethyl cellulose (CMC) and yeast extract as carbon and nitrogen sources, respectively, in a thermo-shaker at 100rpm. The enzyme was partially purified by 80 ammonium sulphate precipitation followed by dialysis, resulting in a 1.52-fold purification. The optimal activity of partially purified cellulase was observed at a temperature of 60C and pH 5. The cellulase enzyme was stable within the pH ranges of 35 and retained 67 of activity even at 55C. Cellulase activity was found to be enhanced in the presence of metal ions such as Cd²⁺, Pb²⁺, and Ba²⁺. The enzyme showed the highest activity when CMC was used as a substrate, followed by cellobiose. The K_m and V_max values of the enzyme were 1.8mg/ml and 10.92μg/ml/min, respectively. The cellulase enzyme obtained from Bacillus licheniformis PANG L had suitable catalytic properties for use in industrial applications.

This study is aimed at isolating and identifying a thermophilic cellulolytic bacterium from hot spring water and characterizing thermostable cellulase produced by the isolate. Enrichment and culture of water sample was used for isolation of bacterial strains and an isolate with highest cellulase activity was chosen for the production, partial purification, and biochemical characterization of the enzyme. Different staining techniques, enzymatic tests, and 16s ribosomal DNA (16s rDNA) gene sequencing were used for the identification of the isolate. The cellulase producing isolate was Gram positive, motile, and sporulated rod‐shaped bacterium growing optimally between 55°C and 65°C. Based on partial 16s rDNA sequence analysis, the isolate was identified as Geobacillus sp. and was designated as Geobacillus sp. KP43. The cellulase enzyme production condition was optimized, and the product was partially purified and biochemically characterized. Optimum cellulase production was observed in 1% carboxymethyl cellulose (CMC) at 55°C. The molecular weight of the enzyme was found to be approximately 66 kDa on 12% SDS‐PAGE analysis. Biochemical characterization of partially purified enzyme revealed the temperature optimum of 70°C and activity over a wide pH range. Further, cellulase activity was markedly stimulated by metal ion Fe^2+. Apart from cellulases, the isolate also depicted good xylanase, cellobiase, and amylase activities. Thermophilic growth with a variety of extracellular enzyme activities at elevated temperature as well as in a wide pH range showed that the isolated bacteria, Geobacillus sp. KP43, can withstand the harsh environmental condition, which makes this organism suitable for enzyme production for various biotechnological and industrial applications.

Visceral Leishmaniasis (VL) is caused by a protozoan parasite Leishmania donovani that is transmitted to humans by an infected female sandfly, Phlebotomus argentipes. VL is common in the Indian sub-continent including Nepal and efforts for its elimination are ongoing. However, expansion of disease towards the higher altitude areas, previously considered as VL free in Nepal, may impact the ability to achieve the elimination target by 2020. This was an exploratory study, where VL suspected patients living exclusively in the non-program districts of Nepal and presenting with fever > 2 weeks and splenomegaly was included. The patients’ blood samples were collected, and DNA was extracted. DNA was subjected to PCR amplification and subsequent sequencing. Additionally, past 10 years data of VL cases from the national databases were analysed to see the trends of the disease in program and non program districts. Analysis of the past 10 years data revealed that trend of VL cases significantly decreased in the program districts (p = 0.001) while it increased in the non-program districts (p = 0.002). The national trend for overall incidence of VL also significantly decreased over this time period. Limited number of patients’ samples (n = 14) were subjected to molecular investigation, and four patients were found to be positive for Leishmania species by PCR. Interestingly, these cases in non-program districts were indeed also L. donovoni complex. All four patients were male with age ranges from 10 to 68 years. GenBank BLAST of the obtained DNA sequences confirmed identified specimens as L. donovani complex. We identified additional VL cases from non-program districts (including the high lands) of Nepal, indicating that the infection could be an emerging threat for the non-program areas of Nepal. The demonstration of VL cases in areas initially considered non-endemic has raised concern about on-going transmission in those regions and may trigger subsequent government plan and action to include those areas in the elimination program. Thus, the government should consider revising the disease control programs to accommodate non-program districts for achieving the VL elimination goal set for 2020.

Background: The Asiatic wild water buffalo (Bubalus arnee) is an endangered species that is conserved in the Koshi Tappu Wildlife Reserve (KTWR), Nepal, and was recently translocated to the Chitwan National Park (CNP). Gastrointestinal (GI) parasites are the cause of significant negative health and production impacts on animals worldwide.Methods: A coprological survey of GI parasites of wild water buffalo was carried out in the CNP in 2020. Fresh dung samples (n = 25) were collected from wild water buffaloes and analysed using sedimentation and flotation techniques for morphological identification of parasite cysts, oocysts and eggs.Results: Nine different GI parasites were recorded of which Entamoeba spp. (20 samples, 80%) were the most common. The presence of Entamoeba spp. was further validated using polymerase chain reaction (PCR) analysis and DNA sequencing. The PCR results were positive for all of the microscopically positive samples, and the species was identified as Entamoeba bovis. Three samples were sequenced and formed a cluster of E. bovis, which was separated from other Entamoeba spp. in phylogenetic analysis.Conclusion: This is the first report for molecular detection of E. bovis from wild water buffaloes in Nepal. Future work should focus on the prevalence of such infections in water buffaloes in forest environments.