Understanding disease burden and transmission dynamics in resource-limited, developing countries like Nepal is often challenging due to a lack of adequate surveillance systems. These issues are exacerbated by limited access to diagnostic and research facilities throughout the country. Nepal has one of the highest COVID-19 case rates (915 cases per 100,000 people) in South Asia, with densely-populated Kathmandu experiencing the highest number of cases. Swiftly identifying case clusters and introducing effective intervention programs is crucial to mounting an effective containment strategy. The rapid identification of circulating SARS-CoV-2 variants can also provide important information on viral evolution and epidemiology. Genomic-based environmental surveillance can help in the early detection of outbreaks before clinical cases are recognized, and identify viral micro-diversity that can be used for designing real-time risk-based interventions. This research aimed to develop a genomic-based environmental surveillance system by detecting and characterizing SARS-CoV-2 in sewage samples of Kathmandu using portable next-generation DNA sequencing devices. Out of 20 selected sites in the Kathmandu Valley, sewage samples from 16 (80%) sites had detectable SARS-CoV-2. A heat-map was created to visualize transmission activity in the community based on viral load intensity and corresponding geospatial data. Further, 41 mutations were observed in the SARS-CoV-2 genome. Some detected mutations (n=9, 2%) were novel and yet to be reported in the global database, with one indicating a frameshift deletion in the spike gene. We also observed more transition than transversion on detected mutations, indicating rapid viral evolution in the host. Our study has demonstrated the feasibility of rapidly obtaining vital information on community transmission and disease dynamics of SARS-CoV-2 using genomic-based environmental surveillance.

Butyrate, a histone deacetylase inhibitor, has several therapeutic applications, including cancer. However, the effect of butyrate in HBV replication is not known so far. It was hypothesized that butyrate might inhibit HBV replication and host cell proliferation via SIRT-1. It was found that the increased expression of SIRT-1 in Hep G2.2.15 cells (HBV expressing cells) than Hep G2 cells. Next the expression of SIRT-1 and Acetylated p53 (Ac-p53) were measured in the liver biopsy samples of chronic hepatitis B (CHB) patients with high viral load and compared to CHB patients with low viral load and found that there was a high SIRT-1 expression and a low Ac-p53 levels in CHB patients with high viral load compared to CHB patients with low viral load. Incubation of butyrate inhibited SIRT-1 expression and cell proliferation. Inhibition of SIRT-1 by butyrate or SIRT-1 siRNA increased the levels of Ac-p53. The elevated Ac-p53 decreased p-akt, cyclin D1, and thereby inhibited cell proliferation. Incubation of butyrate with Hep G2.2.15 cells also inhibited HBx protein expression, HBV-DNA and hepatitis B surface antigen (HBsAg). Taken together, the data showed that butyrate inhibited HBV replication and cell proliferation by inhibiting SIRT-1 expression in hepatoma cells. K E Y W O R D S anti-viral, butyrate, HBV, p53, replication, SIRT-1 1 | INTRODUCTION Hepatitis B Virus (HBV) is a small double stranded-DNA virus, and its infection is a major health problem worldwide. 1 HBV infection is responsible for chronic liver inflammation (hepatitis), which further progresses to liver cirrhosis and liver failure or hepatocellular carcinoma (HCC). 2 The control of HBV replication is one of the most significant approaches to reduce the chronic HBV infection-linked pathogenicity. To date, there is no treatment available which can completely eradicate HBV infection in the patients. Few chemotherapeutic drugs such as, adefovir, entecavir, lamivudine, and telbivudine target the HBV-DNA polymerase and can reduce the viral replication. 3,4 However, long-term use of these drugs might lead to the increase the viral drug-resistance. 4Recently, use of interferon-alpha (IFN-α) along with the analogues of nucleotide/nucleoside significantly minimized the HBV infection and replication in the HBV infected patients. 5,6 Nevertheless, long-term use of IFN-α and nucleotide/nucleoside therapy tends to develop the drug resistance and causes some side effects in the patients. 7,8 Therefore, there is a need for a potential therapy or the targeted drugs for the anti-HBV therapeutics.SIRT-1 or Sirtuin-1 is a class III histone deacetylase (HDAC), which can deactivate histones or many non-histone proteins including p53, NF-κB, AP-1, and PGC-1α. SIRT-1 modulates several cellular processes such as apoptosis, cell cycle regulation, aging, and gene silencing. 9-11 SIRT-1 is also known to induce HBV replication. 12 SIRT-1 inducer resveratrol had been shown to influence the HBV

Dogs are often commensal with human settlements. In areas where settlements are adjacent to wildlife habitat, the management of dogs can affect risk of spillover of disease to wildlife. We assess dog husbandry practices, and measure the prevalence of Canine Distemper Virus (CDV) in dogs, in 10 villages in Nepal’s Annapurna Conservation Area (ACA), an important region for Himalayan wildlife. A high proportion (58%) of owned dogs were allowed by their owners to roam freely, and many village dogs originated from urban areas outside the region. CDV antibodies, indicating past exposure, were detected in 70% of dogs, and 13% were positive for P-gene, suggesting current circulation of CDV. This is the first detection of canine distemper virus in a National Park in Nepal Himalaya. Dogs were generally in good condition, and none exhibited clinical signs of CDV infection, which suggests that infections were asymptomatic. CDV exposure varied with village location and age of dogs, but this variation was minor, consistent with high rates of movement of dogs across the region maintaining high seroprevalence. Residents reported the occurrence of several species of wild carnivores in or close to villages. These results suggest a high potential for transmission of CDV from village dogs to wild carnivores in ACA. We suggest that control of dog immigration, along with vaccination and neutering of dogs could mitigate the risk of CDV spillover into wild carnivore populations.

Poultry (Gallus domesticus) farming plays an important role as an income generating enterprise in a developing country like Nepal, contributing more than 4% to the national GDP. It is also one of the major sources of protein for growing population. Newcastle Disease (ND) is a major poultry disease affecting both commercial and backyard poultry production worldwide. There were more than 90 reported cases of ND outbreaks in Nepal in 2018, with over 74,986 birds being affected. ND might be responsible for over 7% of total poultry mortality in the country. Recent outbreak of ND in 2021 affected many farms throughout Nepal, and caused massive poultry production loss. ND is caused by a single stranded RNA virus which presents very similar clinical symptoms as Influenza A (commonly known as Bird flu), adding much complexity to clinical disease identification and intervention.We conducted a nationwide ND and Influenza A prevalence study, collecting samples from commercial and backyard poultry farms from across the major poultry production hubs of Nepal, and conducted both serological and molecular assessments-giving us disease exposure history and identification of floating strains of ND Virus (NDV). Of 600 commercial chickens tested from various farms, both NDV (n=381, 64%) and IA (n=125, 21%) antibodies were detected in the majority of the samples. In backyard chicken (n=108, 39 farms), sero-prevalence was also relatively high for both NDV (n=38, 35%) and IA (n=17, 16%). Out of the 40 commercial farms, majority had detectable NDV (n=31, 78%) and IA (n=15, 38%) virus present. In backyard farms (n=36), we also detected NDV (n=6, 16%) and IA (n=1, 3%) virus. We Genotyped (strain) detected NDV, and found Genotype II to be present in most of the commercial farms (which might be coming from live vaccine usage) and Genotype I in some backyard poultry samples. The identified Genotype I strain is reported for the first time, and hence could be an endemic NDV strain found in Nepal. Our 2021 ND outbreak investigation identified Genotype VII c as the causative strain.Additionally, we have developed a thermostable I-2 NDV vaccine (Ranigoldunga™) in tablet formulation and tested on various (mixed) breeds of chicken (G. domesticus). This vaccine seems to be highly effective against NDV, including a virulent 2021 outbreak strain (Genotype VII c). The I-2 Tablet ND vaccine showed more than 85% efficacy when administered either ocularly or in water, and has a stability of 30 days in room temperature.

16 Dogs are often commensal with human settlements. In areas where settlements are adjacent to 17 wildlife habitat, the management of dogs can affect risk of spillover of disease to wildlife. 18 We assess dog husbandry practices, and measure the prevalence of Canine Distemper Virus 19 (CDV) in dogs, in 10 villages in Nepal's Annapurna Conservation Area (ACA), an important 20 region for Himalayan wildlife. A high proportion (58%) of owned dogs were allowed by 21 their owners to roam freely, and many village dogs originated from urban areas outside the 22 region. CDV antibodies, indicating past exposure, were detected in 70% of dogs, and 13% 23 were positive for P-gene, suggesting current circulation of CDV. This is the first detection of 2 24 canine distemper virus in a National Park in Nepal Himalaya. Dogs were generally in good 25 condition, and none exhibited clinical signs of CDV infection, which suggests that infections 26 were asymptomatic. CDV exposure varied with village location and age of dogs, but this 27 variation was minor, consistent with high rates of movement of dogs across the region 28 maintaining high seroprevalence. Residents reported the occurrence of several species of wild 29 carnivores in or close to villages. These results suggest a high potential for transmission of 30 CDV from village dogs to wild carnivores in ACA. We suggest that control of dog 31 immigration, along with vaccination and neutering of dogs could mitigate the risk of CDV 32 spillover into wild carnivore populations. 34 Introduction35 Domestic dogs Canis lupus familiaris (hereafter "dogs") have followed the human journey 36 (1) to become the world's most abundant and widely distributed carnivore (2-4). Dog 37 husbandry refers to the selection and management of dogs (through roaming restrictions, feed 38 type and frequency, veterinary care, reproductive management, and so on) to fulfil their 39 assigned function (i.e. pet, herding, guarding). Husbandry shapes the health and fitness of 40 dogs, and affects their behaviour and physiology (5). Through the process of deciding which 41 dogs receive care, which get to reproduce, or even which live or die, humans shape the 42 demography of dog populations (6). Human behaviour also shapes interactions between dogs 43 and wildlife (7). Combined, dog husbandry and human behaviour shape the way dogs 44 respond to pathogens as host or reservoir (8). Studies have investigated the diseases shared 45 between dogs and wildlife (9-11), but there have been few attempts to understand the 46 importance of humans in facilitating the role of dogs as agents of pathogen transmission to 47 wildlife (7,11,12).3 48 49 Canine distemper virus (CDV) is a globally-distributed infectious disease that was once 50 thought to affect dogs only. It has now been detected in over 200 species of terrestrial 51 vertebrates (13,14), and remains an emerging disease because high mutability and subsequent 52 host-switching enables the virus to emerge in previously unknown host species (15)(16)(17).53 Although CDV can ...