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Objectives Community-onset bloodstream infections (BSIs) caused by carbapenemase-producing Enterobacter cloacae complex (ECC) species are increasing internationally. This observation suggests that ECC are emerging pathogens, requiring for detailed understanding on their genomic epidemiology including transmission dynamics and antimicrobial resistance profiles. Patients and methods We performed WGS on 79 Enterobacter spp. isolated from the patients with clinically significant BSIs and admitted to emergency department of a major tertiary hospital in Nepal between April 2016 and October 2017. Results We identified 5 species and 13 STs of ECC. Enterobacter xiangfangensis ST171, one of the globally emerging carbapenem resistant ECC clones with epidemic potential, was the most prevalent (42%). Phylogenetic analysis showed a large (>19 400 SNPs) core genome SNP distance across major STs, which was minimal ( Conclusions Our study highlighted that MDR ECC clones are important pathogens of BSIs in community. Though of low prevalence, carbapenem resistance observed in our ECC isolates raised concern about further community dissemination, underscoring the need for community surveillance to identify MDR ECC clones with epidemic potential.

Background Glanders is a transmissible zoonotic disease caused by Burkholderia mallei that infects equids and humans. No glanders cases in equids were reported so far in Nepal. Case presentation Following suspected glanders in animals with clinical signs in different regions in Nepal, serum samples were tested by CFT, ELISA and Luminex® tests. Two horses and a mule tested positive for glanders by all tests, while two other equids only tested positive by ELISA and Luminex®. Analysis of swabs and pus samples by a PCR system targeting B. mallei confirmed the presence of the bacterium in the samples collected from the 3 equids that yielded positive results in all serological tests. Genotyping of the three PCR positive samples with a SNP-based method identified a genotype closely related to the B. mallei strains circulating in India. Conclusion Confirmation of glanders cases underscores the need of implementing a surveillance program in Nepal and a strict control of the animal movement across the borders.

Bloodstream infections (BSIs) are common cause of morbidity and mortality among patients of all age groups. Of particular concern is the increasing emergence and spread of antimicrobial resistance (AMR) in many Gram-negative bacteria (GNB) that cause BSIs. The aim of this thesis was to determine some of the epidemiological features of BSIs in a tertiary hospital in Nepal providing a specific focus on the molecular aspects of AMR, virulence, and variations in Enterobacter spp., a clinically important GNB associated with BSIs. To better understand BSIs-causing non-Salmonella GNB and their AMR determinants in this setting, I conducted a retrospective surveillance study, accessing data from 2012 to 2018. I identified a high prevalence of multi-drug resistance (MDR) and several key AMR genes associated with predominant GNB pathogens. The neonatal intensive care unit (NICU) and the emergency room (ER) were the most prevalent hospital departments for BSIs. To gain an insight into the epidemiology of hospital-acquired (HA) BSIs and identify potential preventive measures, I conducted a further prospective study in the NICU. An increased isolation of Enterobacter spp. in the NICU during the study period suggested an outbreak, prompting an outbreak investigation of Enterobacter spp. isolates using whole genome sequencing (WGS). Identification of ER to having the highest burden of BSIs due to Enterobacter spp. demanded for an epidemiological research to understand the genomic determinants of virulence, AMR, and variations of Enterobacter spp.. To address this, I performed a comprehensive WGS analysis of Enterobacter spp. isolated in the ER. The conclusion of my work is that BSIs due to antimicrobial resistant non-Salmonella GNB is an expanding problem in our hospital, highlighting the need for a continued AMR surveillance. I found that Enterobacter spp. was an emerging MDR pathogen causing BSIs in this hospital. The complex transmission dynamics of Enterobacter spp. observed in ER warranted for a more comprehensive genomic investigation complemented with demographic and clinical data of patients to better disaggregate the epidemiology of BSIs in ER.

Background: Hearing loss is the most common sensory loss accounting for 250 million population of world out of which 16% is attributed from occupational exposure to loud noise, ranging from 7% to 21% in different parts of the world. The prevalence of noise induced hearing loss is more common among factory workers than general population. There are limited reports regarding occupational noise from developing country like Nepal. Method: A cross sectional study was done in Dharan Industrial Area between February 2014 to May 2014. Twenty different industries operating inside the premises of industrial area were categorized into four major categories as per the nature and types, such as: Metal, Plastic, Food and Chemical based industries. Measurement of the sound was done in two phases. Pure tone audiometry of the exposed and non-exposed population was done. Results: Total 104 workers were included from 20 different factories; out of them 80 workers were exposed to loud noise and 24 were non-exposed who acted as the control. The frequency of hearing loss among exposed population was 38.8% and it was 12.5% among non-exposed and the difference was statistically significant. In exposed population hearing loss was 7.7%, 40.5% and 60.9% in age groups of 15- 30, 31- 45 and > 46 years respectively and the difference was statistically significant. Population working longer duration had more chance of noise induced hearing loss and the difference was significant. Conclusion: Population working in noise producing factory has high risk of developing noise induced hearing loss and the risk increases in older people and in people who works for long duration.

INTRODUCTION Potato, popularly known as king of vegetables, is one of the most important crops of Nepal. It is grown in all agro- ecological zones of 77 districts ranging from 100 to 4400 masl (Dhital & Khatri, 2004). People residing in terai and mid hills consume potato as a major vegetable while in high hills it is used as a staple food. It is the fourth most important crop after rice, wheat and maize; however holds first position in productivity. Its annual production is 2,805,582 tons with the productivity of 14.03 mt/ha (MoAD, 2017). It is highly nutritious food constituting good source of energy, vitamins and minerals. Potato cultivation is a major source of income for small holder farmers in mid and high hills. It plays vital role in maintaining the food security and eliminating the poverty and malnutrition in Nepal. Despite its huge importance and cultivation potentiality, its production is very low as compare to the neighboring countries. Many factors are responsible for low yield of potato but lack of high yielding and late blight resistant varieties is the bottleneck constraint for potato cultivation in Nepal. The production of existing cultivated varieties is not enough to meet increasing market demand. Farmers are forced to recycle the same low yielding variety for years. Variety development is a continuous process to replace old degenerating varieties with new improved varieties. National Potato Research Programme (NPRP) is responsible to conduct series of on-station and on-farm trials for developing new variety. On-farm trials are important to speed up the variety development process and enhance the adoption of new varieties in farm communities (Romney et al., 2003; Assefa et al., 2005). Besides, it enables the incorporation of farmers’ opinions and ensures testing of varieties under farmers’ management. Therefore as a part of on-farm trials, the experiment was conducted to evaluate growth and yield performance of promising potato clones in Bhaktapur and hence improve the productivity of potato through selection of appropriate potato clones in Bhaktapur. These clones were tested in various trials of several stations of the country but in Bhaktapur they were introduced for the first time in the present study. MATERIALS AND METHODS Five promising potato clones PRP 35861.18, CIP 384866.5, PRP 226267.11, CIP388676.1, and PRP 85861.11 were brought from NPRP, Khumaltar, Lalitpur and planted in farmers' field along with Desiree as check variety in winter-spring season of 2018. The experiment was laid out in Randomized Complete Block Design with 4 replications in Bhaktapur (1348 masl). The ridge of 3m long was prepared using potato ridge maker at a spacing of 60 cm. Well sprouted potato tuber seed of 25-30 g containing at least 2 eyes were planted at 25cm apart. The individual plot size was 7.2 m2 with four ridges containing 12 plants each. The plot was fertilized at the rate of 100:100:60kg N:P2O5:K2O and 20mt FYM per hactare. The cultural practices and observation techniques were performed as per the recommendation of NPRP (Khatri et al., 1999). Observations were recorded on growth and yield characters. Plant Uniformity was recorded using 1to 5 scale [1 = Very poor (completely heterogeneous in plant size and vigour), 2 = Poor (Some heterogeneous in plant size and plant vigour), 3 = Fair (average or acceptable), 4 = Good (relatively high homogeneity in plant size, plant vigour and growth) and 5 = Excellent (completely homogeneous in plant size, plant vigour and growth) (Khatri et al., 1999)]. Plant vigour was also recorded using 1 to 5 scale [1 = very weak (all the plants are small, Data entry, management and descriptive analysis was done using MS Excel 2007. ANOVA and DMRT were done using R 3.5.1 and R-Studio 1.1.453.0 version (R core team, 2017) and significance level was defined at 5%. RESULTS Growth characters The results of germination, plant uniformity and vigour are presented in Table 1. Significant variation in germination among the clones was observed at 30 DAP. Desiree showed the higher germination as compared to the promising potato clones of CIP. PRP clones showed similar germination to 'Desiree' indicating competitiveness of new clones to an established early variety. The germination of clones was found statistically non significant at 45 DAP. PRP 85861.11 and PRP 35861.18 were similar but superior to Desiree for plant uniformity at 60 DAE while the variation was non significant at 30 DAE. PRP 35861.18 had higher plant vigor at both 45 DAE and 60 DAE than CIP 388676.1. Table 1. Germination, uniformity and plant vigor of potato clones in Bhaktapur, 2018 Treatment Germination (%) Uniformity Plant Vigor 30DAP 45DAP 45 DAE 60DAE 45DAE 60DAE PRP 35861.18 63.46±12.49a 98.75±2.5 4.5±0.58 4.5±0.58ab 3.75±0.96a 4.25±0.5a CIP 384866.5 30.21±19.65b 92.71±7.12 3.25±0.5 3.5±0.58c 2.75±0.5bc 3.75±0.5ab PRP 226267.11 54.17±36.49ab 93.75±7.98 4.00±0.82 4±0.82abc 3.25±0.5ab 4.25±0.5a CIP 388676.1 34.38±27.72b 90.63±7.12 3.75±0.96 3.75±0.96bc 2.25±0.5c 3.0±0.00c PRP 85861.11 55.13±18.12ab 92.29±3.56 4.25±0.96 5±0.0a 3.25±0.5ab 3.50±0.58bc Desiree 73.75±17.46a 97.92±4.17 3.5±0.56 3.25±0.5c 3±0.0abc 3.00±0.00c F test ** NS NS * * ** P value 0.00002 0.10719 0.25 0.00179 0.0175 0.00163 CV% 30.26 4.65 19.76 16.46 16.89 11.9 LSD 23.65 6.614 1.15 0.99 0.77 0.65 Significance codes: 0 ‘***’ 0.001 ‘**’ 0.01 ‘*’ 0.05 ‘.’ 0.1 ‘ ’ 1 Note: DAP=Days after Planting, DAE= Days after emergence, LSD=Least Significant Difference, CV%=Coefficient of Variation Percentage The results of stem thickness, plant height and ground cover of the potato clones are presented in Table 2. The highest stem thickness was shown with PRP 226267.11. Plant height also varied significantly among the clones. At 45 DAE PRP 35861.18 was the tallest while at 60 DAE PRP 35861.18 and PRP 226267.11 were the tallest among the tested clones. Desiree was the most dwarf clone in both observations. Statistically significant variation was observed in ground cover of clones. PRP 226267.11 and PRP 85861.11 were recorded as superior to Desiree for ground cover. Table 2. Effect of clones on stem thickness, plant height and ground cover in Bhaktapur, 2018 Stem thickness(cm) Plant height (cm) Ground cover at 60 DAE Treatment 45 DAE 60 DAE 45DAE 60DAE PRP 35861.18 3.01±0.22b 3.16±0.06bc 50.95±6.47a 62.45±4.1a 75±8.16ab CIP 384866.5 3.36±0.53ab 3.31±0.2ab 37.80±6.62c 51.16±6.11b 75±4.08ab PRP 226267.11 3.69±0.53a 3.54±0.35a 44.70±6.76b 61.50±11.6a 80±4.08a CIP 388676.1 3.12±0.07b 2.9±0.26c 30.75±3.87d 39.70±5.24c 48.75±4.79c PRP 85861.11 3.27±0.46b 3.08±0.22bc 42.90±6.10b 49.20±3.67b 77.50±6.45a Desiree 3.24±0.26b 3.0±0.23bc 33.63±5.46d 40.88±4.52c 67.5±6.45b P value 0.0281 0.00762 2.84e-07 1.47e-06 3.95e-05 F- test * ** *** *** *** LSD 0.38 0.32 4.15 6.10 9.39 CV% 7.65 6.67 6.86 7.97 8.82 Significance codes: 0 ‘***’ 0.001 ‘**’ 0.01 ‘*’ 0.05 ‘.’ 0.1 ‘ ’ 1 Note: DAE= Days after emergence, LSD=Least Significant Difference, CV%=Coefficient of Variation Percentage Among the 6 tested clones, the growth pattern of PRP 35861.18 was found as lodging, CIP 388676.1 as erect and the remaining 4 clones as spreading type. CIP 388676.1 was categorized as late maturity type (>135), Desiree as early type ( Table 3. Variation in growth characters among potato clones in Bhaktapur, 2018 Treatment Growth Habit Maturity Late Blight PRP 35861.18 Lodging Medium Mild Resistant CIP 384866.5 Spreading Medium Resistant PRP 226267.11 Spreading Medium Resistant CIP 388676.1 Erect Late Highly Resistant PRP 85861.11 Spreading Medium Highly Resistant Desiree Spreading Early Highly Susceptible Yield characters The tuber distribution by weight and number as well as yield of the tested clones is illustrated in Table 4. Significantly higher USN and USW per plant was reported with PRP 85861.11 among the tested clones. SSN and SSW per plant also varied significantly among the clones. PRP 85861.11 produced the highest SSN while PRP 85861.11 and PRP 35861.18 gave the highest SSW per plant. Similarly the significant highest OSN and OSW per plant was observed with CIP 388676.1 Table 4. Effect of clones on yield parameters of potato in Bhaktapur, 2018 Treatments USN / plant USW / plant SSN / plant SSW / Plant OSN/ Plant OSW / plant Yield (mt/ha) PRP 35861.18 3.9 ±1.16b 53.85 ±13.37b 4.95 ±0.74ab 173.25 ±28.94a 1.35 ±0.41b 88.65 ±29.44b 21.05 ±3.20ab CIP 384866.5 6.75 ±2.75b 84.25 ±36.60ab 4.25 ±1.05bc 137.25 ±36.34ab 0.90 ±0.68b 65 ±53.26b 19.1 ±3.35b PRP 226267.11 5.95 ±2.09b 72.00 ±17.76b 4.95 ±1.41ab 168 ±36.34a 1.60 ±0.82b 125.75 ±89.33b 24.38 ±5.49a CIP 388676.1 5.1 ±3.93b 69.1 ±52.51b 3.05 ±1.15c 101.1 ±33.43b 2.55 ±1.0a 196.3 ±93.53a 24.43 ±2.51a PRP 85861.11 10.85 ±0.66a 118.95 ±17.60a 6.2 ±0.75a 174.7 ±16.93a 0.95 ±0.9b 73.90 ±52.81b 24.5 ±4.59a Desiree 4.75 ±1.23b 51.25 ±11.70b 3.95 ±0.19bc 133.35± 16.55ab 1.35 ±0.7b 94 ±48.71b 18.57 ±4.34b F test ** * ** * ** ** * P value 0.0024 0.0455 0.00447 0.0181 0.00713 0.00847 0.02239 CV% 31.61 38.27 20.03 20.01 37.67 41.39 13.20 LSD 2.96 43.20 1.336 43.30 0.81 66.92 4.39 Significance codes: 0 ‘***’ 0.001 ‘**’ 0.01 ‘*’ 0.05 ‘.’ 0.1 ‘ ’ 1 Note: USN=Undersize (50gm) number, USW= Undersize (50gm) weight, LSD=Least Significant Difference, CV%=Coefficient of Variation Percentage The variation in tuber yield of clones was found statistically significant. PRP 85861.11 (24.5mt/ha), CIP 388676.1(24.43mt/ha) and PRP 226267.11(24.38mt/ha) were found statistically similar and superior for tuber yield. Desiree (18.57 mt/ha) produced the least yield among the tested clones. Among the six tested clones, three were oval shaped, two were oblong shaped and one was round in shape. The tubers of 2 clones were with shallow eye, 2 with medium and 2 with deep eyes. Regarding the tuber color, 2 clones were white colored, 3 with red color and one with yellow color. Similarly, variation was noticed in tuber flesh color as mentioned in Table 5. Table 5. Variation in tuber characteristic among potato clones in Bhaktapur, 2018 Treatment Shape Eye depth Tuber color Tuber Flesh color PRP 35861.18 Oval Medium White White CIP 384866.5 Round Deep Yellow Pale yellow PRP 226267.11 Oval Medium Red Cream CIP 388676.1 Oval Shallow White White PRP 85861.11 Oblong Deep Red Cream Desiree Oblong Shallow Red Yellow Based on the external appearance of tuber and the total production, PRP 226267.11 held the first position by farmer’s ranking. Organoleptic taste ranking of boiled potato reported PRP 85861.11 as the most delicious clone among all (Table 6) Table 6. Farmers’ ranking and organoleptic taste ranking Treatment Farmer’s ranking Organoleptic taste ranking of boiled potato PRP 35861.18 3 5 CIP 384866.5 5 2 PRP 226267.11 1 3 CIP 388676.1 2 6 PRP 85861.11 4 1 Desiree 6 4 DISCUSSION The potato clones tested at CFFT, RARS Nepalgunj (NPRP, 2016/17) reported that there is significant variation in germination at 30DAP. The lower germination of promising clones as compared to Desiree might be due to presence of some growth inhibitors in seed tuber and its susceptibility to prolonged frost at early period of planting. Burton and Meigh (1971) also reported that benzothiazole, 1,4-dimethylnaphthalene and 1,6- dimethylnaphthalene are comparatively potent inhibitors of sprout growth in the potato tuber. PRP 35861.18 and CIP 388676.1 had shown the highest and lowest plant vigour respectively in our study which corresponds to the finding of Upadhyay et al. (2017). Ahamad et al. (2012) reported non significant variation in plant uniformity, ground cover and late blight. It might be due to different genotypes, weather and environmental condition. Upadhyay et al. (2017) reported PRP 35861.18 and CIP 388676.1 as the tallest and shortest clones respectively among the tested clones which agrees with our findings. Significant variation in plant height among the genotypes was observed in Gautam et al. (2004) and Ghimire et al. (2004). Abbas (2011) reviewed and reported that varieties vary in flesh and skin colour, eye depth and tuber shape. Upadhyay et al. (2017) reported significant variation in number of seed size tuber and over size tuber only but the finding of this study showed significant variation in number of all tubers types. Cho and Iritani (1983) reported that tuber number is function of stem population but is influenced by cultivars and several other factors, which control vegetative growth. Dhakal et al. (2011) and Chapagain et al. (2014) reported that the varieties significantly influenced the yield of tuber per plant. According to Upadhyay et al. (2017) PRP 85861.11 gave the highest yield and Desiree produced the lowest among 6 tested clones which is quite similar to our finding. It might also be due to similar environmental condition of Bhaktapur and Kavre. CONCLUSION Hence it can be concluded that all 5 promising potato clones performed better than Desiree with respect to growth and yield parameters. PRP 85861.11, CIP 388676.1 and PRP 226267.11 were the superior clones regarding the tuber yield. All the promising clones were found resistant to late blight while Desiree was found highly susceptible. PRP 35861.18 showed the better growth performance. The farmers’ preference was high for PRP 226267.11 and organoleptic taste ranking

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The Eurasian lynx Lynx lynx and the Pallas’s cat Otocolobus manul are elusive felids which are little studied in the Himalayas of Nepal where they seem to occur at low densities. We present mtDNA genomic and dietary data of six Eurasian lynx scats and one Pallas’s cat scat collected in Dolpa district of Nepal. The Eurasian lynx scats were found in alpine shrub lands at elevations from 4,738 - 4,818 m. Dietary analysis based on microscopic frequency of occurrence revealed that the six Eurasian lynx scats contained 56 % woolly hare Lepus oiostolus, 17.7 % pika Ochotona sp. and rodent Alticola sp., 9.7 % Himalayan marmot Marmota himalayana, 7 % vegetation and 3.3 % domestic goat Capra aegagrus hircus. The Pallas’s cat scat found in a rocky boulder field contained 76 % pika hair, 18 % woolly hare hair and 4 % vegetation. This study provides the first scientific record of Eurasian lynx in Shey-Phoksundo National Park SPNP. The finding on Pallas's cat represents an expansion of the IUCN distribution range in Nepal, presents the second locality record of the species in the country, and with the sample collected at 5,539 m is the highest elevation record for the species to date. Further researches into population status, ecology, genetics and conservation of these elusive felids in the Himalayas are recommended.