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.

Statistical optimization of cellulase production from Bacillus sp. YE16 isolated from yak dung of the Sikkim Himalayas for its application in bioethanol production using pretreated sugarcane bagasse.

Cellulolytic bacteria were isolated from yak dung samples collected from different habitats of Sikkim, India. Isolate YE16 from the Yumthang Valley sample showed highest enzyme activity of 7.68 U/mL and was identified as Bacillus sp., which has a sequence similarity of 96.15% with B. velezensis. One factor at a time (OFAT) analysis revealed that an acidic pH of 5 with 37 °C temperature was optimum for maximum enzyme production after 36 hrs of incubation (13.88 U/mL), which was further increased after statistical optimization (34.70 U/mL). Media optimization based on response surface methodology predicted that Carboxymethyl cellulose (CMC) and MgSO4 at concentrations of 30 g/L and 0.525 g/L, respectively, at pH 5.5 to show CMCase activity of 30.612 U/mL, which was consistent with the observed value of 30.25 U/mL and confirmed the model. The crude enzyme also efficiently hydrolyzed alkaline pretreated sugarcane bagasse, releasing 7.09 g/L of glucose equivalent with an ethanol production of 3.05 g.

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.

Climatic variability and its impact on the indigenous agricultural system using panel data analysis in the Sikkim Himalaya, India.

Climate-induced extreme events with fluctuations in climatic indicators like temperature and precipitation highly influence crop productivity. This study deals with quantitative analysis of climatic variability and crop production (1990-2018) using panel data regression analysis. The focus is on variability of three crops, i.e., paddy, maize, and wheat in the Rangit river basin of Sikkim Himalaya, India. Meterological data were acquired from the Indian Meteorological Department, agricultural data from the state agricultural department and a field survey were also conducted with the farmers, using a structured questionnaire, focused group discussion, and key informant observations. The acquired data was analyzed with the help of correlation and multiple linear regression analysis to analyze the relationship between climatic variability and crop production. The result of the study shows that all three crops are dependent on rainfall; however, paddy was the most sensitive to climatic variability. It was found that the overall grain production had an inverse relation to temperature, but it had a positive correlation with rainfall. It was observed that there was a continuous decline in the overall production of paddy and wheat. During deficiency years, it was found that when - 11.33% rainfall was deficient, paddy yield declined by - 1.52%. Further, a deviation of - 13.48% led to a decline of - 54.78% in wheat. The study advocates that timely policy interventions and strategies shall reduce climatic shocks and improve productivity. This would strengthen the livelihood security of the local communities, overcome the challenges of food security, and ensure long-term sustainability of Rangit River basin.

Elevation determines the productivity of large cardamom (Amomum subulatum Roxb.) cultivars in Sikkim Himalaya.

Large cardamom (Amomum subulatum Roxb.) is an economically important cash crop that provides a livelihood option for the rural communities in Sikkim Himalaya. However, its production has declined drastically over the past few decades due to climate change and other factors affecting the livelihood of marginal cardamom-dependent farmers in the region. Climate change causes a shift in elevational distributions of mountain species, and it is pivotal to understand its effect on yield and yield-related traits for economically important plant species like large cardamom. For this, we randomly studied 41 large cardamom cultivation sites in Sikkim (India) with elevations ranging between 975 and 2069 m asl and evaluated the yield-related traits (number of capsules per spike, capsule length, capsule width, fresh capsule weight, dry capsule weight, number of seeds per locule, fresh seed weight, and dry seed weight) in five cultivars (Dzongu Golsey, Sawney, Seremna, Ramsey, and Varlangey). We observed a significant variability (P < 0.05) for morphometric traits in each of the five cultivars cultivated in different elevations. The cultivation of low-elevation cultivars like Seremna and Dzongu Golsey (suitable in elevation < 975 m) has shifted upward to mid (975-1515 m) and high-elevation (> 1515 m), while cultivation of high-elevation Ramsey cultivar (suitable in elevation > 1515 m) has shifted downward (< 1515 m). The Dzongu Golsey, Sawney, and Seremna cultivated in mid-elevation (975-1515 m) performed better in terms of yield-related traits than the same cultivars cultivated in high-elevation (> 1515 m) and showed moderate to high negative correlation between elevation and yield-related traits, indicating the negative effect of elevation on their yield. Likewise, Ramsey and Varlangey cultivated in high elevation (> 1515 m) performed better than the one cultivated in mid-elevation (975-1515 m) and depicted moderate to high positive correlation between elevation and yield-related traits, suggesting a positive influence of elevation on their yield. Although there is an elevational shift in the cultivation of large cardamom cultivars, the elevation influences the performance of the large cardamom cultivars, and it also suggests cultivating the cultivars in their suitable elevation range for better productivity.

Prediction of glaciated area fraction over the Sikkim Himalayan Region, India: a comparative study using response surface method, random forest, and artificial neural network.

Glacier area fraction at high altitude mountains is a serious worry in today's time triggered by climate change. The current information on this natural resource is very important for the survival of humanity as it affects the water, food, and energy security of people dependent on it. Due to its problematic accessibility and tough environmental condition, ground monitoring is quite challenging. This study investigates the impact of environmental parameters and pollutants on glacier area fraction over the Eastern Himalaya region and its prediction through random forest (RF), multilayer perceptron (MLP), radial basis function analysis (RBFN), and response surface methodology (RSM) models. The data are obtained from the Goddard Earth Sciences Data and Information Services Center (GES DISC), NASA's data archive portal ( https://giovanni.gsfc.nasa.gov ). The collinearity of independent variables reveals that all selected input parameters are highly correlated with R2 value > 0.9. The RSM and RF model provided valuable insight of the predictor's significance in addition to their capability to predict the response. The model performance was evaluated in terms of R2 value and the error matrices. The model's R2 value was found to be 0.843, 0.839, 0.838, and 0.743 for MLP, RBFN, RF, and RSM respectively. Although, the neural network model R2 values are the highest, but the most reliable and suitable model is RF as the error matrices for this model are much lower than others. This study encourages the investigation of the hybridization of these models for more accurate prediction.

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.

Characterization of a unique Sikkimese yak population of India: a multivariate approach.

Native Sikkimese yak in Sikkim state of India is a pastoral treasure being raised through centuries-old transhumance practices and has evolved in response to natural and man-made selection. Currently, the population of Sikkimese yak is at risk with about five thousand total headcounts. Characterization is essential for taking appropriate decisions for conservation of any endangered population. In an attempt to phenotypically characterize the Sikkimese yaks, this study recorded phenotypic morphometric traits information, viz., body length (LG), height at withers (HT), heart girth (HG), paunch girth (PG), horn length (HL), horn circumference (HC), distance between horns (DbH), ear length (EL), face length (FL), face width (FW), and tail length with switch (TL), on 2154 yaks of both sexes. Multiple correlation estimation highlighted that HG and PG, DbH and FW, and EL and FW were highly correlated. Using principal component analysis, LG, HT, HG, PG, and HL were found to be the most important traits for phenotypic characterization of Sikkimese yak animals. Discriminant analysis based on different locations of Sikkim hinted at the existence of two separate clusters, however, broadly, phenotypic uniformity could be observed. Subsequent genetic characterization can offer greater insights and can pave the way for future breed registration and conservation of the population.

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.

Characterization of Polycyclic Aromatic Hydrocarbons (PAHs) associated with fine aerosols in ambient atmosphere of high-altitude urban environment in Sikkim Himalaya.

Polycyclic Aromatic Hydrocarbons (PAHs) compounds are ubiquitous in ambient air due to their persistence, carcinogenicity, and mutagenicity. Gangtok being one of the cleanest cities in India located in Eastern Himalayan region, witnesses high developmental activities with enhanced urbanization affecting the ambient air quality. The present study aims to measure PM2.5 and PAHs in the ambient atmosphere of the Sikkim Himalaya to understand the influence of natural and anthropogenic activities on aerosol loading and their chemical characteristics. The PM2.5 samples were collected and analysed for the duration from Jan 2020 to Feb 2021.The seasonal mean concentrations of PM2.5 and PAHs were observed to be high during autumn and low during summer season. Overall, the annual mean concentration of PM2.5 was found higher than the prescribed limit of World Health Organization and National Ambient Air Quality Standards. The concentration of the 16 individual PAHs were found to be highest during autumn season (55.26 ± 37.15 ng/m3). Among the different PAHs, the annual mean concentration of fluorene (3.29 ± 4.07 ng/m3) and naphthalene (1.15 ± 3.76 ng/m3) were found to be the highest and lowest, respectively. The Molecular Diagnostic Ratio (MDR) test reveals higher contribution from heavy traffic activities throughout the winter and autumn seasons. The other possible sources identified over the region are fossil fuel combustion, and biomass burning. The multivariate statistical analysis (Multifactor Principal Component Analysis) also indicates a strong association between PM2.5 /PAHs and meteorological variables across the region in different seasons. The precipitation and wind pattern during the study period suggests that major contribution of the PM2.5 and PAHs were from local sources, with minimal contribution from long-range transport. The findings are important for comprehending the trends of PAH accumulation over a high-altitude urban area, and for developing sustainable air quality control methods in the Himalayan region.

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.

Rahnella sikkimica sp. nov., a novel cold-tolerant bacterium isolated from the glacier of Sikkim Himalaya with plant growth-promoting properties.

The current study describes a novel species with the strain name ERMR1:05T isolated from the forefield soil of East Rathong Glacier in West Sikkim Himalaya (India). The isolate was facultatively anaerobic, gram-stain negative, non-spore-forming, rod-shaped, and oxidase negative. Whole-genome-based bacterial core gene phylogenetic analysis placed the strain in the genus Rahnella, well separated from Rouxiella spp. The digital DNA-DNA hybridisation and average nucleotide identity values between strain ERMR1:05T and other members of genus Rahnella were below the proposed thresholds for the species delineation. Based on these results, a new species, Rahnella sikkimica sp. nov., is proposed with strain ERMR1:05T (CIP 111636T, MTCC 12598T) as the type strain. The bacterium showed upregulation of cold-stress genes in cold conditions. Additionally, the genome analysis of the bacterium showed the presence of plant growth-promotion factors suggesting its role in crop improvement in cold hilly regions.

Monitoring the distribution pattern and invasion status of Ageratina adenophora across elevational gradients in Sikkim Himalaya, India.

Understanding the spread intensity and population dynamics of invasive plant species is a prerequisite for developing management strategies in the Himalayan Forest ecosystems that are experiencing an accelerated rate of climate change. Although there are studies on the occurrence of few invasive species in the Himalayan ecosystems, systematic information on their intensity of spread and species association is still missing. Considering existing data gaps, we aimed to assess the intensity of spread and distribution pattern of A. adenophora, one of the high-concern invasive species (HiCIS) of India that is causing havoc in the Himalayas, across an elevational gradient. Field data were collected in 2018 and 2021 in the Indian federal state of Sikkim, located in the Eastern Himalayas. We analyzed the population status and species association of A. adenophora along an elevational gradient ranging from > 600 m to 2700 m above sea level, which was divided into seven gradients of 300 m width, and each gradient was further randomly sampled. Overall, 81 species were present in association with A. adenophora, including 58 herbs, 19 shrubs, and 4 climbers, belonging to 30 families and 67 genera in the region. No other species continuously co-occurred along with A. adenophora throughout the elevation ranging from > 600 m to 2700 m. The species observed increased frequency (100%), density (40.51 ind./100 m2), and basal cover (11.25 cm2/m2) in the elevational gradient 1500-1800 m in 2018. In 2021, A. adenophora dominated the highest elevational gradient (< 2400-2700 m) with increased frequency (99.96%), density (58.41 ind./100m2), and basal cover (42.54 cm2/100m2), which demonstrated rapid invasion and improved plant health and reproductive vigor in comparison to the lower elevational gradient in Sikkim Himalaya. Despite being completely absent at the highest elevation (< 2400-2700 m), in 2018, it observed gregarious spread at the highest elevation in 2021, which is of serious concern to ecologists. The presence of the targeted species in all seven studied altitudinal gradients reflects stage III of the species invasion. An enormous shift in the distribution pattern along elevational gradients within a short time span is alarming for the Himalayan ecosystem since it is becoming a thriving habitat for invasive species owing to anthropogenic activity. We mapped the potential geographical extent using the species distribution model (SDM) and predicted the suitable habitat of distribution in Sikkim Himalaya. In order to curtail the spread and counteract the negative impact of this species on native vegetation in Sikkim Himalaya and ultimately reverse the process, local and regional initiatives for its biological control and management must be taken.

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.

Description of nine new species of phytoseiid mites (Acari: Mesostigmata) from Sikkim, a north eastern States of India.

A rapid roving survey was done in the East Sikkim district of the state Sikkim with a target to explore phytoseiid fauna of the region. Nine species of phytoseiid mites namely Amblyseius lanceae sp. nov.; Amblyseiulella cancellatus sp. nov.; Amblyseiulella gangtokiensis sp. nov.; Euseius tubuliferus sp. nov.; Okiseius pahari sp. nov.; Paraamblyseius ranipoolensis sp. nov.; Phytoseius ferrum sp. nov.; Typhlodromus (Anthoseius) himaliniae sp. nov. Typhlodromus (Anthoseius) kanchanjanghai sp. nov. are described with proper illustrations in the present paper.

Psychrotrophic plant beneficial bacteria from the glacial ecosystem of Sikkim Himalaya: Genomic evidence for the cold adaptation and plant growth promotion.

Commercial biofertilizers tend to be ineffective in cold mountainous regions due to reduced metabolic activity of the microbial inoculants under low temperatures. Cold-adapted glacier bacteria with plant growth-promoting (PGP) properties may prove significant in developing cold-active biofertilizers for improving mountain agriculture. With this perspective, the cultivable bacterial diversity was documented from the East Rathong glacier ecosystem lying above 3900 masl of Sikkim Himalaya. A total of 120 bacterial isolates affiliated to Gammaproteobacteria (53.33%), Bacteroidetes (16.66%), Actinobacteria (15.83%), Betaproteobacteria (6.66%), Alphaproteobacteria (4.16%), and Firmicutes (3.33%) were recovered. Fifty-two isolates showed many in vitro PGP activities of phosphate solubilization (9-100 µg/mL), siderophore production (0.3-100 psu) and phytohormone indole acetic acid production (0.3-139 µg/mL) at 10 °C. Plant-based bioassays revealed an enhancement of shoot length by 21%, 22%, and 13% in ERGS5:01, ERMR1:04, and ERMR1:05, and root length by 14%, 17%, 11%, and 22% in ERGS4:06, ERGS5:01, ERMR1:04, and ERMR1:05 treated seeds respectively. An increased shoot dry weight of 4-29% in ERMR1:05 and ERMR1:04, and root dry weight of 42-98% was found in all the treatments. Genome analysis of four bacteria from diverse genera predicted many genes involved in the bacterial PGP activity. Comparative genome study highlighted the presence of PGP-associated unique genes for glucose dehydrogenase, siderophore receptor, tryptophan synthase, phosphate metabolism (phoH, P, Q, R, U), nitrate and nitrite reductase, TonB-dependent receptor, spermidine/putrescine ABC transporter etc. in the representative bacteria. The expression levels of seven cold stress-responsive genes in the cold-adapted bacterium ERGS4:06 using real-time quantitative PCR (RT-qPCR) showed an upregulation of all these genes by 6-17% at 10 °C, and by 3-33% during cold-shock, which indicates the cold adaptation strategy of the bacterium. Overall, this study signifies the psychrotrophic bacterial diversity from an extreme glacier environment as a potential tool for improving plant growth under cold environmental stress.

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.

To Study Association of Neck Circumference with Metabolic Syndrome and Cardiovascular Risk Factors - A Hospital Based Case Control Study in Ethnic Population of Sikkim.

Metabolic syndrome is known to increase the risk of cardiovascular diseases, diabetes, stroke. Upper body obesity is found to be associated with metabolic syndrome and increased risk of cardiovascular diseases. This present study is being conducted with the objective to evaluate association of neck circumference with metabolic syndrome in ethnic population of Sikkim. MATERIAL: The present study is a hospital based pilot study conducted at Dept. of Medicine at a tertiary care centre in Northeast India after ethical clearance from institute's ethical committee and after taking written informed consent, A total of 30 subjects had been evaluated for components of metabolic syndrome and cardiovascular risk factors. OBSERVATION: 30 subjects of metabolic syndrome were screened, raised triglyceride level was most commonly found, Neck circumference was found to be statistically significant associated with metabolic syndrome, 56.6 % subject with metabolic syndrome had abnormal neck circumference. CONCLUSION: Upper body obesity had been recognized as to increase risk of cardiovascular disease and neck circumference can be used as simple and novel indicator. Patients with larger neck circumference should be screened for cardiovascular risk factors.

In vitro and genetic screening of probiotic properties of lactic acid bacteria isolated from naturally fermented cow-milk and yak-milk products of Sikkim, India.

A total of 272 isolates of lactic acid bacteria (LAB) were isolated from 22 samples of naturally fermented milk products of Sikkim in India viz. dahi, soft-variety chhurpi, hard-variety chhurpi, mohi and philu, out of which, 68 LAB isolates were randomly grouped on the basis of phenotypic characteristics, and were identified by 16S rRNA gene sequence analysis. Leuconostoc mesenteroides was the most dominant genus, followed by Leuc. mesenteroides subsp. jonggajibkimchii, Lactococcus lactis subsp. cremoris, Lc. lactis, Lc. lactis subsp. hordniae, Lc. lactis subsp. tructae, Enterococcus faecalis, E. italicus and E. pseudoavium. LAB strains were tested for probiotics attributes by in vitro and genetic screening, based on marker genes. LAB strains showed tolerance to pH 3.0, bile salt, resistance to lysozyme and ß-galactosidase activity. Enterococcus faecalis YS4-11 and YS4-14 and Lactococcus lactis subsp. cremoris SC3 showed more than 85% of hydrophobicity. Genes clp L and tdc encoding for low pH tolerance, agu A and Ir1516 encoding for bile tolerance, LBA1446 gene encoding for BSH activity, map A, apf, mub 1 and msa encoding for mucosal binding property were detected. Gene mesY for bacteriocin production was detected only in Leuconostoc spp. Based on the in vitro and genetic screening of probiotic attributes, Leuc. mesenteroides; Leuc. mesenteroides subsp. jonggajibkimchii and Lc. lactis subsp. cremoris were tentatively selected for possible probiotic candidates.

Multilocus sequence based identification and adaptational strategies of Pseudomonas sp. from the supraglacial site of Sikkim Himalaya.

Microorganisms inhabiting the supraglacial ice are biotechnologically significant as they are equipped with unique adaptive features in response to extreme environmental conditions of high ultraviolet radiations and frequent freeze-thaw. In the current study, we obtained eleven strains of Pseudomonas from the East Rathong supraglacial site in Sikkim Himalaya that showed taxonomic ambiguity in terms of species affiliation. Being one of the most complex and diverse genera, deciphering the correct taxonomy of Pseudomonas species has always been challenging. So, we conducted multilocus sequence analysis (MLSA) using five housekeeping genes, which concluded the taxonomic assignment of these strains to Pseudomonas antarctica. This was further supported by the lesser mean genetic distances with P. antarctica (0.73%) compared to P. fluorescens (3.65%), and highest ANI value of ~99 and dDDH value of 91.2 of the representative strains with P. antarctica PAMC 27494. We examined the multi-tolerance abilities of these eleven Pseudomonas strains. Indeed the studied strains displayed significant tolerance to freezing for 96 hours compared to the mesophilic control strain, while except for four strains, seven strains exhibited noteworthy tolerance to UV-C radiations. The genome-based findings revealed many cold and radiation resistance-associated genes that supported the physiological findings. Further, the bacterial strains produced two or more cold-active enzymes in plate-based assays. Owing to the polyadaptational attributes, the strains ERGC3:01 and ERGC3:05 could be most promising for bioprospection.