Chrono-spatial reproductive phenology of Swertia bimaculata Hook.f. & Thomson ex C.B.Clarke reveals suitability as climate-changing bio-indicator along the Sikkim Himalayan altitude.

Genus Swertia under the family Gentianaceae is morphologically and taxonomically distinct with about 14 species from Sikkim Himalayan region. A Chrono-spatial floral phenology study was conducted on Swertia bimaculata from 1580 to 2400 m altitude in West Sikkim Himalayan region. The floral phenology was studied in a field nursery at 1200 m altitude and spatial floral phenology was studied over a 1580-2400 m altitudinal range in the West Sikkim Himalayan region. Detailed study on flowering pattern and seed formation in S. bimaculata revealed late flowering at 1200 m compared to 2400 m. Plants at the lower altitude showed prolonged and variable flowering from September to November and seed formation from November to December. During September to mid-October, pentamerous and bisexual flowers were observed, followed by 9-16 % tetramerous until November; and up to 24 % trimerous and 10 % bimerous until the end of flowering season in December. By the end of December, 43 % of flowers become stamen-less (monoclinous). Less than 2 % seed formation was observed in stamen-less flowers. The temporal phenological behavior reveals a successful reproductive adaptive strategy, depicting pentamerous flowers during favorable conditions, reducing to tetramerous, followed by tri- & bimerous with female (monoclinous) flowers until the end of flowering with stressful conditions, suggesting its suitability as a stress indicator to the changing climatic conditions. Along the altitudinal range floral anomalies from the usually observed pentamerous (and very rare hexamerous) flowers in favorable conditions change to tetramerous and/or trimerous, as the environmental conditions become more stressful, suggesting that the changes in this species could be used as an indicator of stress in changing climatic conditions.

Population dynamics and genetic isolation in the Central Himalayan region: insights from Sikkim population, India.

India's rich diversity encompasses individuals from varied geographical, cultural, and ethnic backgrounds. In the field of population genetics, comprehending the genetic diversity across distinct populations plays a crucial role. This study presents significant findings from genetic data obtained from the Sikkimese population of India. Autosomal markers were crucial for evaluating forensic parameters, with a combined paternity index of 1 × 109. Notably, Penta E emerged as a distinguishing marker for individual identification in the Sikkim population. Fst genetic distance values revealed insights into genetic isolation among different groups, enhancing our understanding of population dynamics in the central Himalayan region. The NJ-based phylogenetic tree highlighted close genetic relationships, of the Sikkim population with the Nepalese population surrounding neighbouring Himalayan populations providing glimpses into common ancestry. In summary, this study contributes valuable data to population genetics and underscores the importance of genetic variation in comprehending population dynamics and forensic applications.

Unique genetic architecture of prolificacy in 'Sikkim Primitive' maize unraveled through whole-genome resequencing-based DNA polymorphism.

KEY MESSAGE: 'Sikkim Primitive' maize landrace, unique for prolificacy (7-9 ears per plant) possesses unique genomic architecture in branching and inflorescence-related gene(s), and locus Zm00001eb365210 encoding glycosyltransferases was identified as the putative candidate gene underlying QTL (qProl-SP-8.05) for prolificacy. The genotype possesses immense usage in breeding high-yielding baby-corn genotypes. 'Sikkim Primitive' is a native landrace of North Eastern Himalayas, and is characterized by having 7-9 ears per plant compared to 1-2 ears in normal maize. Though 'Sikkim Primitive' was identified in the 1960s, it has not been characterized at a whole-genome scale. Here, we sequenced the entire genome of an inbred (MGUSP101) derived from 'Sikkim Primitive' along with three non-prolific (HKI1128, UMI1200, and HKI1105) and three prolific (CM150Q, CM151Q and HKI323) inbreds. A total of 942,417 SNPs, 24,160 insertions, and 27,600 deletions were identified in 'Sikkim Primitive'. The gene-specific functional mutations in 'Sikkim Primitive' were classified as 10,847 missense (54.36%), 402 non-sense (2.015%), and 8,705 silent (43.625%) mutations. The number of transitions and transversions specific to 'Sikkim Primitive' were 666,021 and 279,950, respectively. Among all base changes, (G to A) was the most frequent (215,772), while (C to G) was the rarest (22,520). Polygalacturonate 4-α-galacturonosyltransferase enzyme involved in pectin biosynthesis, cell-wall organization, nucleotide sugar, and amino-sugar metabolism was found to have unique alleles in 'Sikkim Primitive'. The analysis further revealed the Zm00001eb365210 gene encoding glycosyltransferases as the putative candidate underlying QTL (qProl-SP-8.05) for prolificacy in 'Sikkim Primitive'. High-impact nucleotide variations were found in ramosa3 (Zm00001eb327910) and zeaxanthin epoxidase1 (Zm00001eb081460) genes having a role in branching and inflorescence development in 'Sikkim Primitive'. The information generated unraveled the genetic architecture and identified key genes/alleles unique to the 'Sikkim Primitive' genome. This is the first report of whole-genome characterization of the 'Sikkim Primitive' landrace unique for its high prolificacy.

Education Intervention on 'Hearing Impairment - Its Assessment and Management' among Medical Practitioners in Sikkim, India.

BACKGROUND: Health care providers must be aware of the choices in aural rehabilitation methods and assessment procedures available presently to those with hearing impairment in order to facilitate early identification and intervention and ensure quality of care to those hearing impaired individuals. The referrals from the medical fraternities for the audiological services are not uniform across India. Part of reason may be that awareness about the roles of and responsibilities of audiologist's is poor among medical practitioners. In some regions of India medical practitioners may be only grossly be aware of audiologists per se. They may not know enough to refer all individuals with hearing or balance disorders for the required audiological services. AIM: To evaluate changes in pre survey and post survey after education intervention specifically planned for medical practitioners in the state of Sikkim, India. Education intervention included awareness about hearing impairment and its assessment and management by audiologists. METHOD: A pre and post survey comparison research design with purposive convenient sampling technique was applied. Participants were medical practitioners working in geographical area of Sikkim at the time of the study. Inspection of demographic data of respondents showed that they were in their age range of 24 and 60 years and with work experience ranging 1-40 years. Participants were invited for an awareness talk which included, information about hearing loss, early identification and early intervention, tests and management of hearing loss in different age groups, need for hearing aids/cochlear implants, auditory training, and the role of an audiologist in the management of hearing loss and ways to prevent hearing loss. A pre and post awareness program responses were collected from all the participants using a customized questionnaire tool. RESULTS: Respondents exhibited, in the pre-survey questionnaire, lack of awareness in many aspects of audiology, including assessing hearing impairment, diagnosing hearing loss, and the role of an audiologist. There was an overall, statistically significant difference in the level of performance on the pre-awareness and post-awareness responses. CONCLUSION: Following an awareness campaign among the medical professionals in Sikkim, there was a considerable change in their level of awareness of hearing impairment, its assessment, and management by audiologists. The reach of the awareness campaign was significant as around 70% of subjects showed significant change in their knowledge and attitude towards hearing impairment, its management.

Anticancer effects of rutin from Fagopyrum tataricum (tartary buckwheat) against osteosarcoma cell line.

BACKGROUND: The present study is analysisof the seeds of buckwheat (Fagopyrum sp.),member of the Polygonaceae family for isolation of rutin and its anticancer property againstOsteosarcoma celllines (SAOS2). The selected plant is traditionally used for diabetes and cancer. It has several biological properties such as antibacterial, antioxidant and anti-aging. PURPOSE: Thirty-five buckwheat cultivars were obtained from Nepal Agriculture Genetic Resources Centre (NAGRC) Khumaltar, Kathmandu, Nepal, and Kumrek Sikkim. These plant varieties are scientifically evaluated their biological properties. METHODS: Rutin wasfractionated from buckwheat seeds using methanol fraction and analysed for quality by HPLC method. The rutin fraction of the cultivar NGRC03731 a tartary buck wheat and standard rutin was used against Osteosarcoma cell lines (SAOS2) and human gingival fibroblast cells (hGFs) for anticancer activity. The cell viability using rutin fraction and standard rutin treated with SAOS2 cells were assessed by MTT assay. For further research, the best doses (IC-50: 20 g/ml) were applied. By using AO/EtBr dual staining, the effects of Rutin fraction on SAOS2 cell death were analysed. The scratch wound healing assay was used to analyse cell migration. Real-time PCR was used to analyse the pro-/anti-apoptotic gene expression. RESULTS: The seeds with the highest rutin content, NGRC03731 seeds, had 433 mg/100 g of rutin.The rutin fraction treatment and standard rutin significantly reduced cell viability in the MTT assay, and osteosarcoma cells were observed on sensitive to the IC-50 dose at a concentration of 20 g/ml after 24 h.The SAOS2 cells exposed to rutin fraction at 20 g/ml and standard rutin at 10 g/ml exhibited significant morphological alterations, cell shrinkage and decreased cell density, which indicate apoptotic cells.Rutin-fraction treated cells stained with acridine orange/ethidium bromide (AO/EtBr) dual staining cells turned yellow, orange, and red which indicatesto measure apoptosis.The anti-migration potential of rutin fraction, results prevented the migration of SAOS2 cancer cells.Rutin-fraction significantly increased the expression of pro-apoptotic proteinsBad, using real-time PCR analysis (mRNA for Bcl-2 family proteins) resulted Bcl-2's expression is negatively regulated. CONCLUSION: Osteosarcoma (SAOS2) cell lines' proliferation, migration, and ability to proliferate were reduced markedly by rutin fraction and it also causes apoptosis of Osteosarcoma cell lines (SAOS2).

Anti-rheumatic potential vis-à-vis aconitine and hypaconitine content analysis in different Aconitum spp. from Sikkim Himalayas (India).

Aconitum spp. are important medicinal plants mentioned in Ayurveda as Ativisa or Vatsanabha. The present study aims to evaluate anti-rheumatic potential in seven Aconitum species and correlation with aconitine and hypaconitine content. Anti-rheumatic potential was analyzed through in vitro xanthine oxidase inhibition, anti-inflammatory and ROS scavenging assays; and quantification of aconitine and hypaconitine with RP-HPLC method validated as per ICH guidelines. The findings reveal that A. palmatum possessed the most promising response (IC50 =12.68±0.15 µg/ml) followed by A. ferox (IC50 =12.912±1.87 µg/ml) for xanthin oxidase inhibition. We observed a wide variation in aconitine and hypaconitine content ranging from 0.018 %-1.37 % and 0.0051 %-0.077 % respectively on dry weight basis. Aconitine and hypaconitine showed moderate positive correlation (r=0.68 and 0.59 respectively) with anti-rheumatic potential. The study identifies potential alternative species of Aconitum that can help in sustainable availability of quality raw material.

Vaccine Breakthrough Infections Among Healthcare Workers in a COVID-19-Designated Tertiary Care Government Hospital in Sikkim.

Introduction Since the emergence of the coronavirus disease 2019 (COVID-19) virus at the beginning of 2020, the world has gone through various waves of pandemics. The health care workers (HCWs) or the COVID warriors as they were termed were the first line of defense against the virus. They were armed with personal protective equipment and prophylactic doses of the COVID-19 vaccine. Despite these precautions, some of the HCWs still contracted the disease and a few others succumbed to it. The objective of this study was to estimate the prevalence of COVID-19 infections and vaccine breakthrough infections (BTIs) in HCWs after receiving the COVID-19 vaccine during the second wave of the pandemic. Methods This was a cross-sectional, hospital-based study conducted over a period of four months from September 2021 to December 2021 on HCWs aged 18 years and above working at the COVID-19-designated tertiary care government hospital in Sikkim. A structured coded questionnaire with no patient identifiers was used to gather details on demographics, vaccination history, breakthrough infection, and other social details. HCWs who had received at least one dose of the COVID-19 vaccine at the time of initiation of the study and were >18 years of age were included in this study. Results A total of 678 HCWs were screened, out of which 229 (33%) participants tested positive for COVID-19 and the rest of the participants (455; 67%) tested negative. COVID-19 infections and vaccine BTIs (COVID-19 infection >14 days after the second vaccination) were recorded and 137 (20%) respondents had a post-vaccination COVID-19 infection out of which 115 (18.5%) were BTI. The majority of the participants were females and of the age group of 26-35 years. The correlation of COVID-19 infections with the dose gap between vaccination, gender, age, profession, department, area posted during COVID duty, cycles of duty performed, hospitalization due to infection, influenza vaccination, and comorbidity was analyzed. Conclusion COVID-19 vaccines are disease-modifying and they decrease the severity of BTIs in HCWs. Pandemics and outbreaks cannot be predicted; therefore, it becomes very important to have healthy frontline workers who are constantly exposed to infectious agents. Monitoring of health and surveillance of infectious diseases among the HCWs should be encouraged.

Phytoconstituents from Urtica dioica (stinging nettle) of Sikkim Himalaya and their molecular docking interactions revealed their nutraceutical potential as α-amylase and α-glucosidase inhibitors.

In this study, antioxidative methanolic leaf extract (MeOH-SIS) of Urtica dioica was characterized for anti-diabetic activity. The extract was purified on a column to yield seven homogenous fractions (F1-F7) which were further determined for DPPH radical scavenging activity. MeOH-SIS and the fraction F1 (selected based on % yield and activity) were evaluated for their in vitro α-amylase and α-glucosidase inhibitory activity. The results showed inhibition of both enzymes in a dose dependent manner and F1 exhibited relatively higher inhibition than its mother extract MeOH-SIS. GC-MS analyses of both the extracts identified 24 major compounds among which 10 were previously described as bioactive compounds. Among all, 5 compounds demonstrated to have quality pharmacokinetics profiles and were examined for possible binding affinity against the active sites of α-amylase and α-glucosidase using molecular docking. The binding interaction of 2R-acetoxymethyl-1,3,3-trimethyl-4 T-(3-methyl-2-buten-1-yl)-1 T-cyclohexanol within the active sites of the target receptors was found to be significant among others, and can be developed as a potential inhibitor of α-amylase and α-glucosidase. The leaf extract can be utilized to develop food additive for the control and management of oxidative stress induced diabetes.

GIS-based data-driven bivariate statistical models for landslide susceptibility prediction in Upper Tista Basin, India.

Predicting landslides is becoming a crucial global challenge for sustainable development in mountainous areas. This research compares the landslide susceptibility maps (LSMs) prepared from five GIS-based data-driven bivariate statistical models, namely, (a) Frequency Ratio (FR), (b) Index of Entropy (IOE), (c) Statistical Index (SI), (d) Modified Information Value Model (MIV) and (e) Evidential Belief Function (EBF). These five models were tested in the high landslides-prone humid sub-tropical type Upper Tista basin of the Darjeeling-Sikkim Himalaya by integrating the GIS and remote sensing. The landslide inventory map consisting of 477 landslide locations was prepared, and about 70% of all landslide data was utilized for training the model, and 30% was used to validate it after training. A total of fourteen landslide triggering parameters (elevation, slope, aspect, curvature, roughness, stream power index, TWI, distance to stream, distance to road, NDVI, LULC, rainfall, modified fournier index, and lithology) were taken into consideration for preparing the LSMs. The multicollinearity statistics revealed no collinearity problem among the fourteen causative factors used in this study. Based on the FR, MIV, IOE, SI, and EBF approaches, 12.00%, 21.46%, 28.53%, 31.42%, and 14.17% areas, respectively, identified in the high and very high landslide-prone zones. The research also revealed that the IOE model has the highest training accuracy of 95.80%, followed by SI (92.60%), MIV (92.20%), FR (91.50%), and EBF (89.90%) models. Consistent with the actual distribution of landslides, the very high, high, and medium hazardous zones stretch along the Tista River and major roads. The suggested landslide susceptibility models have enough accuracy for usage in landslide mitigation and long-term land use planning in the study area. Decision-makers and local planners may utilise the study's findings. The techniques for determining landslide susceptibility can also be employed in other Himalayan regions to manage and evaluate landslide hazards.

Baseline metagenome-assembled genome (MAG) data of Sikkim hot springs from Indian Himalayan geothermal belt (IHGB) showcasing its potential CAZymes, and sulfur-nitrogen metabolic activity.

Here we present the construction and characterization of metagenome assembled genomes (MAGs) from two hot springs residing in the vicinity of Indian Himalayan Geothermal Belt (IHGB). A total of 78 and 7 taxonomic bins were obtained for Old Yume Samdong (OYS) and New Yume Samdong (NYS) hot springs respectively. After passing all the criteria only 21 and 4 MAGs were further studied based on the successful prediction of their 16 S rRNA. Various databases were used such as GTDB, Kaiju, EzTaxon, BLAST XY Plot and NCBI BLAST to get the taxonomic classification of various 16 S rRNA predicted MAGs. The bacterial genomes found were from both thermophilic and mesophilic bacteria among which Proteobacteria, Chloroflexi, Bacteroidetes and Firmicutes were the abundant phyla. However, in case of OYS, two genomes belonged to archaeal Methanobacterium and Methanocaldococcus. Functional characterization revealed the richness of CAZymes such as Glycosyl Transferase (GT) (56.7%), Glycoside Hydrolase (GH) (37.4%), Carbohydrate Esterase family (CE) (8.2%), and Polysaccharide Lyase (PL) (1.9%). There were negligible antibiotic resistance genes in the MAGs however, a significant heavy metal tolerance gene was found in the MAGs. Thus, it may be assumed that there is no coexistence of antibiotic and heavy metal resistance genes in these hot spring microbiomes. Since the selected hot springs possess good sulfur content thus, we also checked the presence of genes for sulfur and nitrogen metabolism. It was found that MAGs from both the hot springs possess significant number of genes related to sulfur and nitrogen metabolism.

Surface area change detection of snow/ice cover over Zemu Glacier of Sikkim Himalaya.

This study assessed the snow/ice surface area changes over the Zemu Glacier in the Eastern Himalayas. Zemu is considered to be the largest glacier in the Eastern Himalayas located in the Sikkim State of India. Change detection in the snow/ice surface areal extent of the Zemu Glacier was delineated from the year 1945 using US Army Map Service-Topographical Sheets and Landsat imageries available from 1987 to 2020. The results obtained solely focus on surface-change delineation using remote sensing satellite data and GIS software. Landsat imagery of the years 1987, 1997, 2009, 2018, and 2020 was used for snow and ice pixel extraction. Normalized Difference Snow Index (NDSI), Snow Cover Index (S3), and new band ratio index were used to extract the pure snow and ice pixels, fresh snow, and debris-covered snow/ice area and pixels mixed with shadow to demarcate the surface area changes. Manual delineation was required and undertaken for better results. A slope raster image was generated from Shuttle Radar Topography Mission (SRTM) digital elevation model (DEM) data to delineate the slope and hill shade. The results illustrate that the snow/ice surface covering the glacier had an area of 113.5 km2 in the year 1945 which was reduced to 78.31 km2 in the year 2020 indicating a total snow/ice surface area loss of 31% in 75 years. A loss of 11.45% was observed from 1945 to 1987 in the areal extent, while from 1987 to 2009, a decadal loss of approximately 7% was observed. The surface area loss of 8.46% from 2009 to 2018 led to infer the maximum loss of snow and ice over the glacier body at a rate of 0.94% per year. Between the years 2018 and 2020, the glacier lost 1.08% of its surface area. The Accumulation Area Ratio (AAR) considering the accumulation and ablation areas of the glacier indicated a gradual decline of the accumulation zone in the recent years. Global Land Ice Measurement from Space (GLIMS) program data with RGI version 6.0 was used as a reference to demarcate the areal extent of Zemu Glacier. The study gained above 80% of overall accuracy by generating a confusion matrix in ArcMap. The analysis of this seasonal snow/ice cover showed that there has been a drastic decline in the surface snow/ice cover area over the Zemu Glacier over the years 1987 to 2020 and NDSI; S3 analysis provided improved results to delineate snow/ice cover area over such steep terrain topography of Sikkim Himalaya.

Antioxidant prodigiosin-producing cold-adapted Janthinobacterium sp. ERMR3:09 from a glacier moraine: Genomic elucidation of cold adaptation and pigment biosynthesis.

Janthinobacterium from cold niches has been studied broadly for bioactive violacein production. However, reports on the atypical red-pigmented Janthinobacterium strains are shallow. The bioactive red prodigiosin pigment has immense pharmacological significance, including antioxidant, antimicrobial and anticancer potential. Here, we report the first complete genome of a prodigiosin-producing Janthinobacterium sp. ERMR3:09 from Sikkim Himalaya in an attempt to elucidate its cold adaptation and prodigiosin biosynthesis. Nanopore sequencing and Flye assembly of the ERMR3:09 genome resulted in a single contig of 6,262,330 bp size and 62.26% GC content. Phylogenomic analysis and genome indices indicate that ERMR3:09 is a potentially novel species of the genus Janthinobacterium. The multicopy cold-responsive genes and gene upregulation under cold stress denoted its cold adaptation mechanisms. Genome analysis identified the unique genes, gene cluster and pathway for prodigiosin biosynthesis in ERMR3:09. Considering the notable antioxidant activity, it can be the next powerhouse of bioactive prodigiosin production.

Post-monsoon seasonal variation of prokaryotic diversity in solfataric soil from the North Sikkim hot spring.

The solfataric soil sediments of the hot springs of Sikkim located at Yume Samdung and Lachen valley were studied for deciphering the bacterial diversity. The main aim here is to present a comparative study and generate a baseline data on the post-monsoon seasonal variation for the months of October and December, analyzed through 16S rRNA V3-V4 amplicon sequencing. The results have shown that there is not much variation at phylum level in the month of October in all the three hot springs such as New Yume Samdung (NYS), Old Yume Samdung (OYS), and Tarum (TAR) hot spring. The abundant phyla mainly present were Firmicutes, followed by Proteobacteria, Actinobacteria, and Bacteroidetes. Similarly, in the month of December, Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetes were prevalent; however, the percent relative abundance of these phyla in the month of December is relatively less. Besides this decrease in percent abundance, it was interestingly seen that relatively more phyla were found contributing towards the bacterial diversity in the month of December. Similar to phylum level, at genus level, there was not much variation seen among various prevalent genera of the three studied hot springs in both months. The major genera prevalent in both months among all the three hot springs were followed by Bacillus, Desulfotomaculum, Lactobacillus, and Paenibacillus. A similar trend was also seen at gene level that relative abundance of various genera was higher in the month of October but more genera were found to be contributing towards bacterial diversity in the month of December. Few distinct genera were found to be more abundant in the month of December such as Rhodopirellula and Blastopirellula. The results may conclude that there is not much variation in the abundance and type of bacterial communities during the post-monsoon season in the month of October and December. However, this may be assumed that there is the accumulation or increase in the bacterial communities during the winter (relatively higher temperature among hot springs) and may favor few mesophilic and more thermophilic communities as well.

Moderate resolution LAI prediction using Sentinel-2 satellite data and indirect field measurements in Sikkim Himalaya.

The leaf area index (LAI) has been traditionally used as a photosynthetic variable. LAI plays an essential role in forest cover monitoring and has been identified as one of the important climate variables. However, due to challenges in field sampling, complex topography, and availability of cloud-free optical satellite data, LAI assessment on larger scale is still unexplored in the Sikkim Himalayan area. We used two optical instruments, digital hemispherical photography (DHP) and LAI-2200C, to assess the LAI across four different forests following 20 × 20 m2 elementary sampling units (ESUs) in the Himalayan state of Sikkim, India. The use of Sentinel-2 derived vegetation indices (VIs) demonstrated a better correlation with the DHP based LAI estimates than using LAI-2200C. Further, the combination of both reflectance bands and VIs were integrated to predict the LAI maps using random forest model. The temperate evergreen forests demonstrated the highest LAI value, while the predicted maps exhibited LAI maxima of 3.4. The estimated vs predicted LAI for DHP and LAI-2200C based estimation demonstrated reasonably good (R2 = 0.63 and R2 = 0.68, respectively) agreement. Further, improvements on the LAI prediction can be attempted by minimizing errors from the inherent field protocols, optimizing the density of field measurements, and representing heterogeneity. The recent rise of frequent forest fires in Sikkim Himalaya prompts for better understanding of fuel load in terms of surface fuel or canopy fuel that can be linked to LAI. The high-resolution LAI map could serve as input to forest fuel bed characterization, especially in seasonal forests with significant variations in green leaves and litter, thereby offering inputs for forest management in changing climate.

Developing ecotourism sustainability maximization (ESM) model: a safe minimum standard for climate change mitigation in the Indian Himalayas.

Recently, ecotourism has been identified as an adaptation strategy for mitigating climate change impacts, as it can optimize carbon sequestration, biodiversity recovery, and livelihood benefits and generate new opportunities for the sustenance of the economy, environment, and society of the area endowed with natural resources and cultural values. With the growing responsibility at the global level, ecotourism resource management (ERM) becomes inevitable for its sustainable requirements. The integration of ecological and socio-economic factors is vital for ERM, as has been demonstrated by developing an Ecotourism Sustainability Maximization Model for an area under study, that is the Yuksam-Dzongri corridor (also known as Kangchendzonga Base Camp Trek), in the Khangchendzonga Biosphere Reserve (KBR), Sikkim, India. This model is based on the earlier developed ecotourism sustainability assessment (ESA) framework by the authors, which is based on the hierarchical relationship among ecotourism principles, criteria, indicators, and verifiers. Employing such relationships, this paper attempts to maximize ecotourism sustainability (ES) as a function of its sustainability principles, criteria, indicators, and verifiers, subject to the constraints identified through the safe minimum standard (SMS) approach by employing linear programming. Using 58 indicators as decision variables and 114 constraints, the model resulted in a maximum level of achievable ES with a score of 84.6%, allowing the resultant optimum values of the indicators to be maintained at the operational level. A central tenet of the model is the collective responsibility and adoption of a holistic approach involving the government, tourists, tourism enterprises, and local people.

Political stability and its effect on economy: some lessons from Sikkim Himalaya.

The Himalayan state of Sikkim has a unique political culture and social environment that could be replicated. Unlike other north-eastern states of India where ethnic identities have been intensely used for electoral politics and state developmentalism, the collective Sikkimese identity has taken precedence over narrow ethnic identity politics in Sikkim. As a result, the state has evolved as one of the most politically stable and better-governed states in India, resulting in rapid economic growth. This has been made possible by the charismatic quality of its political leadership, and the peoples' collective vision of an inclusive Sikkimese identity.

Coexistence of Heavy Metal Tolerance and Antibiotic Resistance in Thermophilic Bacteria Belonging to Genus Geobacillus.

Hot springs are thought to be potential repositories for opportunistic infections, such as antibiotic-resistant strains. However, there is a scarcity of information on the mechanisms of antibiotic resistance gene (ARG) uptake, occurrence, and expression in thermophilic bacteria. Furthermore, because the genesis and proliferation of ARGs in environmental microorganisms are unknown, the research on antibiotic resistance profiles and probable mechanisms in thermophilic bacteria will become increasingly important. The goals of this study are to explore bacterial diversity, antibiotic and heavy metal resistance, and the prevalence and presence of ARG and metal resistance gene (MRG) in Geobacillus species. The 16S rRNA sequencing was used to determine the culturable bacterium diversity of 124 isolates. Standard Kirby Bauer Disc Diffusion and tube dilution procedures were used to determine antibiotic sensitivity and minimum inhibitory concentration (MIC). The tube dilution method was also used to check metal tolerance. To detect ARG and heavy MRG (HMRG), whole genome sequencing studies of the type species of the genus Geobacillus and five randomly selected Geobacillus species were performed. Graph Pad Prism and XLSTAT were used to perform statistical analyses such as ANOVA, EC50 analysis, and principal component analysis (PCA). The phylum Firmicutes and the genus Geobacillus dominated the culture-dependent bacterial diversity. Surprisingly, all thermophilic isolates, i.e., Geobacillus species, were sensitive to at least 10 different antibiotics, as evidenced by the lack of ARGs in whole genome sequencing analysis of numerous Geobacillus species. However, some of these isolates were resistant to at least five different heavy metals, and whole genome sequencing revealed the presence of MRGs in these thermophilic bacteria. The thermophilic genus Geobacillus is generally antibiotic sensitive, according to this study. In contrast, heavy metal is tolerated by them. As a result, it is possible that ARGs and MRGs do not coexist in these bacteria living in hot springs.

Characterization of ACE inhibitory and antioxidant peptides in yak and cow milk hard chhurpi cheese of the Sikkim Himalayan region.

In this study, simulated in vitro GI digestion of the Himalayan hard chhurpi cheese resulted in the increase of hydrolyzed protein content, antioxidant and ACE-inhibitory activities. LC-MS/MS-based peptidomics revealed a total of 1473 peptides in the samples originating from different milk proteins, including α-S1-casein, α-S2-casein, ß-casein, κ-casein, α-lactalbumin, and ß-lactoglobulin, out of which 60 peptides have been reported for different functional properties. A total of 101 peptides were predicted to be antihypertensive using the bioactivity prediction web servers, AHTpin and mAHTPred. In silico molecular docking studies predicted 20 antihypertensive peptides, exhibiting non-bond interactions between hard chhurpi peptides and ACE catalytic residues. A peptide, SLVYPFPGPI, identified in GI digested cow hard chhurpi and undigested, and GI digested samples of yak hard chhurpi, showed a stronger binding affinity towards ACE. Identifying antioxidant and ACE inhibitory peptides in hard cheese products adds value to them as functional foods of the Himalayan region.

Antioxidant Potential of Selected Wild Edible Leafy Vegetables of Sikkim Himalayan Region: Effects of Cooking Methods and Gastrointestinal Digestion on Activity.

Green leafy vegetables or GLVs are one of the main attractions in the local vegetable market and are widely consumed as the main course and side dish in the Sikkim Himalayan region (SHR). This study evaluated the total phenolic (TPC) and flavonoid contents (TFC) and antioxidant potential in different extracts such as methanolic (MeOH), ethyl acetate (EtOAC), and hexane extracts of selected GLVs followed by changes in the antioxidant activity on cooking and stimulated gastrointestinal (GI) digestion. The MeOH extracts of Urtica dioica L. (Sisnu), Nasturtium officinale W. T. Aiton (Simrayo), Diplazium esculentum Retz. Sw. (Ningro), and Chenopodium album L. (Bethu) were estimated to have higher TPC [22.73-45.84 µg gallic acid equivalent (GAE)/mg of extract]. In contrast, the plant extracts prepared using EtOAC (except for N. officinale, where TFC was found to be higher in hexane extract) were found to contain higher TFC (3.42-14.86 µg quercetin equivalent (QE)/mg of extract). The MeOH extracts also exhibited higher 2, 2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity (9.55-18.67 µg ascorbic acid equivalent (AAE)/mg of extract), total antioxidant activity (TAA) (0.27-0.32 mg AAE/mg of extract), and reducing power potential (RPP) (1.6-9.9 µg AAE/mg of extract). Among the test MeOH extracts, U. dioica demonstrated relatively higher antioxidant activities and was selected for cooking experiments followed by simulated GI digestion. The findings revealed that the loss of antioxidant activity was minimal in steam-cooked leaves (3.5% in 40 min) as compared to the boiled ones (18% in 10 min). The simulated GI (simulated salivary, gastric, and intestinal) digestion performed on raw, steam cooked, and boiled U. dioica leaves showed substantial enhancement of antioxidant properties (by 64.63%) through steam cooking in comparison to the raw leaves. Overall the study concludes that higher antioxidant properties can be achieved on the consumption of steam-cooked U. dioica leaves.

Cold-adaptive traits identified by comparative genomic analysis of a lipase-producing Pseudomonas sp. HS6 isolated from snow-covered soil of Sikkim Himalaya and molecular simulation of lipase for wide substrate specificity.

The genomic analysis of industrially important bacteria can help in understanding their capability to withstand extreme environments and shed light on their metabolic capabilities. The whole genome of a previously reported broad temperature active lipase-producing Pseudomonas sp. HS6, isolated from snow-covered soil of the Sikkim Himalayan Region, was analyzed to understand the capability of the bacterium to withstand cold temperatures and study its lipolytic nature. Pseudomonas sp. HS6 was found to be psychrotolerant with an optimal growth temperature ranging between 25 and 30 °C, with the ability to grow at 5 °C. The genome harbours various cold-adaptation genes, such as cold-shock proteins, fatty acid alteration, and cold stress-tolerance genes, supporting the psychrotolerant nature of the organism. The comparative analysis of Pseudomonas sp. HS6 genome showed the presence of amino acid substitutions in genes that favor efficient functioning and flexibility at cold temperatures. Genome mining revealed the presence of four triacylglycerol lipases, among which the putative lipase 3 was highly similar to the broad temperature-active lipase purified and characterized in our previous study. In silico studies of putative lipase 3 revealed broad substrate specificity with partial and no inhibition of the enzyme activity in the presence of PMSF and orlistat. The presence of genes associated with cold adaptations and true lipases with activity at broad temperature and substrate specificity in the genome of Pseudomonas sp. HS6 makes this bacterium a suitable candidate for industrial applications.