Symptoms 6 months following SARS-CoV-2 infection in Nepali women.

In Nepal, over 1 million individuals have tested positive for SARS-CoV-2. We sought to describe the frequency of nonrecovery from this infection at 6 months and associated symptoms. We conducted a retrospective cohort study of 6142 women who had positive and negative PCR tests for this infection 6 months previously at 3 institutions in Kathmandu. In telephone interviews women provided information on 22 symptoms and their intensities, health status and history, and functional status. Of 3732 women who had tested PCR positive, 630 (16.9%) reported that they were unrecovered. These 630 unrecovered women were distinguished statistically from the 3102 recovered women by more frequent histories of allergies, rheumatoid disease, BCG immunization, Covid vaccination, strep throat and recent URIs, and both weight gain and weight losses of more than 5 kg in the 6 months following testing, and stressful events in the preceding year. Fatigue, pain, difficulty remembering, shortness of breath, heat and cold intolerance and unrefreshing sleep were reported in 41.9% to 10.5% of these 630 unrecovered women. Six months after confirmed SARS-CoV-2 infection 16.9% of Nepali women have long-COVID manifested as an immune, metabolic, and hormonal systems disruptive and dysfunction syndrome.

Assessment of Antibiotic Resistance among Clinical Isolates of Enterobacteriaceae in Nepal.

Clinicians face a global challenge treating infections caused by Enterobacteriaceae because of the high rate of antibiotic resistance. This cross-sectional study from the Nepal Armed Police Force Hospital, Kathmandu, Nepal, characterized resistance patterns in Enterobacteriaceae across different antimicrobial classes and assessed incidences of multidrug-resistant (MDR) and extensively drug-resistant (XDR) infections. Enterobacteriaceae from clinical samples were isolated on blood and MacConkey agar, except for urine samples on cysteine lactose electrolyte-deficient agar. To determine antimicrobial susceptibility patterns, including MDR and XDR, the Kirby-Bauer disc diffusion method was used. Statistics were performed using SPSS, v. 17.0. Members of the family were identified in 14.5% (95% CI: 16.2-12.8%) of the total samples (N = 1,617), primarily in urine (54.7%, 128/234), blood (19.7%, 46/234), and sputum (15.0%, 35/234). Escherichia coli (n = 118, 44.2%) was the most predominant bacteria, followed by Citrobacter freundii (n = 81, 30.3%). As much as 95.6% (392/410) of the isolates were penicillin-resistant, whereas only 36.2% (290/801) were carbapenem-resistant. A total of 96 (36.0%) MDR and 98 (36.7%) XDR Enterobacteriaceae were identified. Proteus mirabilis (44.4%, 8/18) predominated MDR cultures, whereas C. freundii (53.1%, 43/81) predominated XDR cultures. Multidrug resistant (38.4%, 71/154) and XDR Enterobacteriaceae (22.7%, 35/154) were chiefly uropathogens. Fluoroquinolone resistance rates in non-MDR, MDR, and XDR isolates were 19.9%, 63.2%, and 96.2%, respectively, whereas cephalosporin resistance rates were 28.6%, 72.9%, and 95.4% and penicillin resistance rates were 67.0%, 97.4%, and 98.0%. One-seventh of patients visiting the hospital were found to be infected with Enterobacteriaceae, and of these patients, at least one-fourth were infected with MDR strains.

Seroprevalence of SARS-CoV-2 infection in the general population of Nepal during the first and second generalized waves of the COVID-19 pandemic-2020-2021.

Few seroprevalence studies have been conducted on coronavirus disease (COVID-19) in Nepal. Here, we aimed to estimate seroprevalence and assess risk factors for infection in the general population of Nepal by conducting two rounds of sampling. The first round was in October 2020, at the peak of the first generalized wave of COVID-19, and the second round in July-August 2021, following the peak of the wave caused by the delta variant of SARS-CoV-2. We used cross-sectional probability-to-size (PPS)-based multistage cluster sampling to estimate the seroprevalence in the general population of Nepal at the national and provincial levels. We tested for anti-SARS-CoV-2 total antibody using the WANTAI SARS-CoV-2 Ab ELISA kit. In Round 1, the overall national seroprevalence was 14.4%, with provincial estimates ranging from 5.3% in Sudurpaschim to 27.3% in Madhesh Province. In Round 2, the estimated national seroprevalence was 70.7%, with the highest in the Madhesh Province (84.8%) and the lowest in the Gandaki Province (62.9%). Seroprevalence was comparable between males and females (Round 1, 15.8% vs. 12.2% and Round 2, 72.3% vs. 68.7%). The seroprevalence in the ecozones-Terai, hills, and mountains-was 76.3%, 65.3%, and 60.5% in Round 2 and 17.7%, 11.7%, and 4.6% in Round 1, respectively. In Nepal, COVID-19 vaccination was introduced in January 2021. At the peak of the first generalized wave of COVID-19, most of the population of Nepal remained unexposed to SARS-CoV-2. Towards the end of the second generalized wave in April 2021, two thirds of the population was exposed.

Assessment of four in vitro phenotypic biofilm detection methods in relation to antimicrobial resistance in aerobic clinical bacterial isolates.

INTRODUCTION: The lack of standardized methods for detecting biofilms continues to pose a challenge to microbiological diagnostics since biofilm-mediated infections induce persistent and recurrent infections in humans that often defy treatment with common antibiotics. This study aimed to evaluate diagnostic parameters of four in vitro phenotypic biofilm detection assays in relation to antimicrobial resistance in aerobic clinical bacterial isolates. METHODS: In this cross-sectional study, bacterial strains from clinical samples were isolated and identified following the standard microbiological guidelines. The antibiotic resistance profile was assessed through the Kirby-Bauer disc diffusion method. Biofilm formation was detected by gold standard tissue culture plate method (TCPM), tube method (TM), Congo red agar (CRA), and modified Congo red agar (MCRA). Statistical analyses were performed using SPSS version 17.0, with a significant association considered at p<0.05. RESULT: Among the total isolates (n = 226), TCPM detected 140 (61.95%) biofilm producers, with CoNS (9/9) (p<0.001) as the predominant biofilm former. When compared to TCPM, TM (n = 119) (p<0.001) showed 90.8% sensitivity and 70.1% specificity, CRA (n = 88) (p = 0.123) showed 68.2% sensitivity and 42% specificity, and MCRA (n = 86) (p = 0.442) showed 65.1% sensitivity and 40% specificity. Juxtaposed to CRA, colonies formed on MCRA developed more intense black pigmentation from 24 to 96 hours. There were 77 multi-drug-resistant (MDR)-biofilm formers and 39 extensively drug-resistant (XDR)-biofilm formers, with 100% resistance to ampicillin and ceftazidime, respectively. CONCLUSION: It is suggested that TM be used for biofilm detection, after TCPM. Unlike MCRA, black pigmentation in colonies formed on CRA declined with time. MDR- and XDR-biofilm formers were frequent among the clinical isolates.

Analysis of altered level of blood-based biomarkers in prognosis of COVID-19 patients.

INTRODUCTION: Immune and inflammatory responses developed by the patients with Coronavirus Disease 2019 (COVID-19) during rapid disease progression result in an altered level of biomarkers. Therefore, this study aimed to analyze levels of blood-based biomarkers that are significantly altered in patients with COVID-19. METHODS: A cross-sectional study was conducted among COVID-19 diagnosed patients admitted to the tertiary care hospital. Several biomarkers-biochemical, hematological, inflammatory, cardiac, and coagulatory-were analyzed and subsequently tested for statistical significance at P<0.01 by using SPSS version 17.0. RESULTS: A total of 1,780 samples were analyzed from 1,232 COVID-19 patients (median age 45 years [IQR 33-57]; 788 [63.96%] male). The COVID-19 patients had significantly (99% Confidence Interval, P<0.01) elevated levels of glucose, urea, alanine transaminase (ALT), aspartate aminotransaminase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), white blood cell (WBC), C-reactive protein (CRP), procalcitonin (PCT), interleukin-6 (IL-6), ferritin, D-Dimer, and creatinine phosphokinase-MB (CPK-MB) compared to the control group. However, the levels of total protein, albumin, and platelets were significantly (P<0.01) lowered in COVID-19 patients compared to the control group. The elevated levels of glucose, urea, WBC, CRP, D-Dimer, and LDH were significantly (P<0.01) associated with in-hospital mortality in COVID-19 patients. CONCLUSIONS: Assessing and monitoring the elevated levels of glucose, urea, ALT, AST, ALP, WBC, CRP, PCT, IL-6, ferritin, LDH, D-Dimer, and CPK-MB and the lowered levels of total protein, albumin, and platelet could provide a basis for evaluation of improved prognosis and effective treatment in patients with COVID-19.

COVID-19 among Patients Visiting the Department of Emergency of a Tertiary Care Centre: A Descriptive Cross-sectional Study.

Introduction: Because of the unbridled transmissibility of the SARS-CoV-2 worldwide, researchers and healthcare professionals have set a common goal for timely diagnosis and future prevention of the disease. The aim of this study was to find out the prevalence of COVID-19 among patients visiting the Department of Emergency of a tertiary care centre. Methods: This descriptive cross-sectional study was conducted among the individuals suspected COVID-19 who had visited the Department of Emergency of a tertiary care centre between 11 January 2021 and 29 December 2021. Ethical approval was taken from Ethical Review Board (Reference number: 2768). Socio-demographic details, clinical symptoms, and two nasopharyngeal swab samples (one in viral transport medium to run RT-PCR and the other for Ag-RDT) were collected from each individual. Convenience sampling method was used. Point estimate and 95% Confidence Interval were calculated. Results: Among the 232 patients, COVID-19 was detected in 108 (46.55%) (40.13-52.97, 95% CI) by Ag-RDT. A total of 44 (39.63%) of age groups 31-40 years were predominantly infected with SARS-CoV-2. The mean age was 32.13±10.80 years and was mostly males 73 (65.77%). Fever was present in 57 (51.35%) and dry cough was present in 50 (45.05%) COVID-19 patients. Conclusions: The prevalence of COVID-19 among hospitalized individuals in this study was higher than in previous studies conducted in similar settings. Keywords: COVID-19; Nepal; prevalence; SARS-CoV-2.

Human Papillomavirus among Women Undergoing Papanicolaou Smear Test in the Department of Gynaecologic Oncology of a Tertiary Care Centre.

Introduction: In invasive cervical specimens or precursors, high-risk human papillomavirus Deoxyribonucleic acid may be detected to identify females at risk of developing cervical cancer. This study aimed to find out the prevalence of human papillomavirus among women undergoing Papanicolaou smear tests in a tertiary care centre. Methods: A descriptive cross-sectional study was conducted among women undergoing the Papanicolaou smear test in the Department of Gynaecologic Oncology, Nepal Armed Police Force Hospital, between 1 June 2022 and 15 November 2022. Ethical approval was obtained from the Ethical Review Board. A convenience sampling method was used. The point estimate was calculated at a 95% Confidence Interval. Results: Among the 199 women, 6 (3.02%) (0.64-5.40, 95% Confidence Interval) had human papillomavirus infection. The mean age of the infected females was 31.17±5.57 years. Human papillomavirus DNA for 16 and 18 were detected in 4 (66.67%) and 2 (33.33%) females, respectively. Conclusions: The prevalence of human papillomavirus in females was found to be lower than other studies done in similar settings. Keywords: cytology; histology; human papillomavirus; Nepal; prevalence.

Co-infection of Uropathogenic Escherichia coli among COVID-19 Patients Admitted to a Tertiary Care Centre: A Descriptive Cross-sectional Study.

Introduction: Simultaneous infection of antibiotic-resistant uropathogens in patients with COVID-19 has necessitated the revision of the prescription of broad-spectrum antibiotics on the grounds of evidence-based studies and antimicrobial stewardship principles. The objective of this study was to find out the prevalence of uropathogenic Escherichia coli co-infection among hospital-admitted COVID-19 patients of a tertiary care centre. Methods: This descriptive cross-sectional study was conducted in urinary tract infection suspected COVID-19 patients admitted to a tertiary care hospital, from 25th June to 24th December 2021 after ethical clearance from the Institutional Review Committee with registration number 207707860. Convenience sampling was used. Serum procalcitonin levels were also measured. Data analysis was performed using the Statistical Package for the Social Sciences software version 17.0. Point estimate at 95% Confidence Interval was calculated along with frequency and proportion for binary data, and mean and standard deviation for continuous data. Results: Among the 49 hospital-admitted COVID-19 patients, 3 (6.12%) (0.59-12.83 at 95% Confidence Interval) were co-infected with uropathogenic Escherichia coli. Absolute non-susceptibility of Escherichia coli to antibiotics such as ceftriaxone, cotrimoxazole, nalidixic acid, gentamicin, and ampicillin was observed. All isolates were multidrug-resistant. All co-infected patients were female and had a median age of 35 years. Mean±SD value for procalcitonin in patients with co-infection (6.13±7.88 ng/ml) was six times higher than for the patients without co-infection (0.95±1.11 ng/ml). Conclusions: Escherichia coli co-infection in hospitalised COVID-19 patients was less frequent as compared to published literature. The serum procalcitonin value in patients with co-infection was substantially higher than that of patients without co-infection. Keywords: antimicrobial drug resistance; co-infection; COVID-19; Escherichia coli; procalcitonin.

Stress Fractures among Paramilitary Trainee Visiting a Paramilitary Hospital of Nepal: A Descriptive Cross-sectional Study.

INTRODUCTION: Young paramilitary recruits, who undergo strenuous exercise during basic training, are often presented with stress fractures, which could be due to an inadequate vitamin D (25-hydroxyvitamin D) intake. This study aimed to find the prevalence of stress fracture among young paramilitary trainees visiting the orthopedic outpatient department of a paramilitary hospital. METHODS: This was a descriptive cross-sectional study done among paramilitary trainees in a paramilitary Hospital of Nepal between April 2019 to April 2021. The study was approved by the Ethical Review Board (Reference number: 1003) of the Nepal Health Research Council. Convenience sampling was used. Anthropometric variables, serum 25-hydroxyvitamin D level, and bone mineral density of spine and hip were determined. Data analysis was performed using the Statistical Package for the Social Sciences software version 17.0. Point estimate at 95% Confidence Interval was calculated along with frequency, proportion for binary data and mean, standard deviation for continuous data. RESULTS: Among 417 young paramilitary trainees, 24 (5.76%) (3.52-7.99 at 95% Confidence Interval) were found to have a stress fracture. The stress fracture patients had a serum 25-hydroxyvitamin D level of 21.47ng/mL±6.98. Similarly, the bone mineral density value of the spine and hip among the patients was -1.34g/cm2±1.37 and 0.36g/cm2±1.24, respectively. CONCLUSIONS: The prevalence of stress fracture among young paramilitary trainee was high compared to previous studies. Additionally, average Vitamin D and the bone mineral density value of the spine and the total hip among stressed fractured patients were also low.

Role of Tocilizumab in the Treatment of COVID-19 Patients with Cytokine Storm: A Case Series.

The in-hospital mortality in patients with COVID-19 could be correlated with severe acute respiratory syndrome coronavirus-2 induced hyper-inflammation, which is attributed to an unconstrained inflammatory cytokine storm. The pro-inflammatory cytokine, specifically, interleukin-6 plays a prominent role in the cytokine storm and may result in alveolar-capillary blood-gas exchange dysfunction. Therefore, the method to block the signal transduction pathway of interleukin-6 could be a potential treatment for severe COVID-19 patients. In this case series of three patients with severe COVID-19, we focus on the rationale for utilization of tocilizumab, an anti-interleukin-6 receptor antibody, which could block the signal transduction pathway of interleukin-6. The observations from this study allowed us to hypothesize that the infusions of tocilizumab may not reduce the elevated level of interleukin-6, and hence may not be a significant therapeutic for reducing in-hospital mortality associated with COVID-19. Additionally, it could also be speculated that interleukin-6 may not be a potentially actionable target cytokine to treat COVID-19-associated cytokine storms. Keywords: COVID-19; cytokines; interleukin-6.

Barriers and facilitators to the implementation of a national multisectoral action plan for the prevention and control of noncommunicable diseases in Nepal: perspectives of stakeholders.

BACKGROUND: Nepal adopted the Multisectoral Action Plan for the Prevention and Control of Non-Communicable Diseases (MSAP) in 2014. Implementation of the plan has been challenging, with limited participation from non-health sectors. OBJECTIVES: The overall aim of the study was to gain the perspectives of key stakeholders involved in the Nepal MSAP on the barriers and facilitators to its implementation, through the participation of relevant sectors in the plan. METHODS: We held face-to-face semi-structured interviews with 12 stakeholders working in sectors involved in the MSAP. These sectors included the Office of the Prime Minister and Council of Ministries; Ministry of Health and Population (MOHP); Ministry of Education, Science and Technology; Ministry of Forest and Environment; academia; and professional organizations. Thematic analysis of transcripts was used to identify themes on awareness of NCDs, awareness of the MSAP, and barriers and facilitators to participation in the MSAP. RESULTS: Participants recognised NCDs as a growing and major burden in Nepal. However, a number of participants were not familiar with the MSAP, identifying a lack of leadership and poor dissemination. Political and systemic transformation, since the adoption of the MSAP, was seen as a key barrier to implementation. International commitments to develop multisectoral action made by the Government of Nepal were identified as drivers. The recent establishment of a separate section for NCDs and Mental Health within the Department of Health Services of MOHP and the promotion of a Health in All Policies (HiAP) approach in recent national documents, were both considered to support implementation. CONCLUSIONS: The establishment of permanent multisectoral or multistakeholder mechanisms has been challenging despite strong political calls for their development. Moving beyond 2020, multisectoral action plans should engage with stakeholders from federal, provincial and local governments in order to develop costed action plans with specific roles and responsibilities for each sector.

Metallo-ß Lactamase Producing Non-Fermentative Gram-Negative Bacilli from Various Clinical Isolates in a Tertiary Care Hospital: A Descriptive Cross-sectional Study.

INTRODUCTION: Non-fermentative gram-negative bacilli are common causes of human infections especially nosocomial infections. These organisms are usually resistant to multiple antimicrobial agents including carbapenems. The study aimed to find out the prevalence of metallo-ß-lactamase producing non-fermentative gram-negative bacilli among the samples which yielded growth of bacteria in a tertiary care hospital. METHODS: This is a descriptive cross-sectional study conducted in a tertiary care hospital from February 2017 to May 2017. Convenience sampling method was used. Bacterial identification, characterization and antimicrobial susceptibility testing were done by following standard microbiological guidelines. Metallo-ß-lactamase production was detected by using combined disk diffusion test and double-disc synergy test. Data were analyzed by using Statistical Package of Social Science software version 16. Point estimate at 95% confidence interval was calculated along with frequency and proportion for binary data. RESULTS: Among 628 samples which yielded growth of bacteria, 118 (18.79%) at 95% Confidence Interval (15.74-21.84) were metallo-ß-lactamase producing non-fermentative gram-negative bacilli. Among them, 54 (45.76%) were Pseudomonas aeruginosa and 64 (54.24%) were Acinetobacter baumannii. CONCLUSIONS: A high prevalence of metallo-ß-lactamase production was observed among the nonfermentative gram-negative bacilli than the study done in similar settings. It is mandatory to perform routine monitoring of metallo-ß-lactamase producing isolates in clinical laboratories in order to help the clinicians prescribe proper antibiotics.

Effect of the COVID-19 pandemic response on intrapartum care, stillbirth, and neonatal mortality outcomes in Nepal: a prospective observational study.

BACKGROUND: The COVID-19 pandemic response is affecting maternal and neonatal health services all over the world. We aimed to assess the number of institutional births, their outcomes (institutional stillbirth and neonatal mortality rate), and quality of intrapartum care before and during the national COVID-19 lockdown in Nepal. METHODS: In this prospective observational study, we collected participant-level data for pregnant women enrolled in the SUSTAIN and REFINE studies between Jan 1 and May 30, 2020, from nine hospitals in Nepal. This period included 12·5 weeks before the national lockdown and 9·5 weeks during the lockdown. Women were eligible for inclusion if they had a gestational age of 22 weeks or more, a fetal heart sound at time of admission, and consented to inclusion. Women who had multiple births and their babies were excluded. We collected information on demographic and obstetric characteristics via extraction from case notes and health worker performance via direct observation by independent clinical researchers. We used regression analyses to assess changes in the number of institutional births, quality of care, and mortality before lockdown versus during lockdown. FINDINGS: Of 22 907 eligible women, 21 763 women were enrolled and 20 354 gave birth, and health worker performance was recorded for 10 543 births. From the beginning to the end of the study period, the mean weekly number of births decreased from 1261·1 births (SE 66·1) before lockdown to 651·4 births (49·9) during lockdown-a reduction of 52·4%. The institutional stillbirth rate increased from 14 per 1000 total births before lockdown to 21 per 1000 total births during lockdown (p=0·0002), and institutional neonatal mortality increased from 13 per 1000 livebirths to 40 per 1000 livebirths (p=0·0022). In terms of quality of care, intrapartum fetal heart rate monitoring decreased by 13·4% (-15·4 to -11·3; p<0·0001), and breastfeeding within 1 h of birth decreased by 3·5% (-4·6 to -2·6; p=0·0032). The immediate newborn care practice of placing the baby skin-to-skin with their mother increased by 13·2% (12·1 to 14·5; p<0·0001), and health workers' hand hygiene practices during childbirth increased by 12·9% (11·8 to 13·9) during lockdown (p<0·0001). INTERPRETATION: Institutional childbirth reduced by more than half during lockdown, with increases in institutional stillbirth rate and neonatal mortality, and decreases in quality of care. Some behaviours improved, notably hand hygiene and keeping the baby skin-to-skin with their mother. An urgent need exists to protect access to high quality intrapartum care and prevent excess deaths for the most vulnerable health system users during this pandemic period. FUNDING: Grand Challenges Canada.

Trends for Neonatal Deaths in Nepal (2001-2016) to Project Progress Towards the SDG Target in 2030, and Risk Factor Analyses to Focus Action.

INTRODUCTION: Nepal has made considerable progress on improving child survival during the Millennium Development Goal period, however, further progress will require accelerated reduction in neonatal mortality. Neonatal survival is one of the priorities for Sustainable Development Goals 2030. This paper examines the trends, equity gaps and factors associated with neonatal mortality between 2001 and 2016 to assess the likelihood of Every Newborn Action Plan (ENAP) target being reached in Nepal by 2030. METHODS: This study used data from the 2001, 2006, 2011 and 2016 Nepal Demographic and Health Surveys. We examined neonatal mortality rate (NMR) across the socioeconomic strata and the annual rate of reduction (ARR) between 2001 and 2016. We assessed association of socio-demographic, maternal, obstetric and neonatal factors associated with neonatal mortality. Based on the ARR among the wealth quintile between 2001 and 2016, we made projection of NMR to achieve the ENAP target. Using the Lorenz curve, we calculated the inequity distribution among the wealth quintiles between 2001 and 2016. RESULTS: In NDHS of 2001, 2006, 2011 and 2016, a total of 8400, 8600, 13,485 and 13,089 women were interviewed respectively. There were significant disparities between wealth quintiles that widened over the 15 years. The ARR for NMR declined with an average of 4.0% between 2001 and 2016. Multivariate analysis of the 2016 data showed that women who had not been vaccinated against tetanus had the highest risk of neonatal mortality (adjusted odds ratio [AOR] 3.38; 95% confidence interval [CI] 1.20-9.55), followed by women who had no education (AOR 1.87; 95% CI 1.62-2.16). Further factors significantly associated with neonatal mortality were the mother giving birth before the age of 20 (AOR 1.76; CI 95% 1.17-2.59), household air pollution (AOR 1.37; CI 95% 1.59-1.62), belonging to a poorest quintile (AOR 1.37; CI 95% 1.21-1.54), residing in a rural area (AOR 1.28; CI 95% 1.13-1.44), and having no toilet at home (AOR 1.21; CI 95% 1.06-1.40). If the trend of neonatal mortality rate of 2016 continues, it is projected that the poorest family will reach the ENAP target in 2067. CONCLUSIONS: Although neonatal mortality is declining in Nepal, if the current trend continues it will take another 50 years for families in the poorest group to attain the 2030 ENAP target. There are different factors associated with neonatal mortality, reducing the disparities for maternal and neonatal care will reduce mortality among the poorest families.

A conceptual framework for understanding illegal killing of large carnivores.

The growing complexity and global nature of wildlife poaching threaten the survival of many species worldwide and are outpacing conservation efforts. Here, we reviewed proximal and distal factors, both social and ecological, driving illegal killing or poaching of large carnivores at sites where it can potentially occur. Through this review, we developed a conceptual social-ecological system framework that ties together many of the factors influencing large carnivore poaching. Unlike most conservation action models, an important attribute of our framework is the integration of multiple factors related to both human motivations and animal vulnerability into feedbacks. We apply our framework to two case studies, tigers in Laos and wolverines in northern Sweden, to demonstrate its utility in disentangling some of the complex features of carnivore poaching that may have hindered effective responses to the current poaching crisis. Our framework offers a common platform to help guide future research on wildlife poaching feedbacks, which has hitherto been lacking, in order to effectively inform policy making and enforcement.

First isolation of dengue virus from the 2010 epidemic in Nepal.

Dengue is an emerging disease in Nepal and was first observed as an outbreak in nine lowland districts in 2006. In 2010, however, a large epidemic of dengue occurred with 4,529 suspected and 917 serologically-confirmed cases and five deaths reported in government hospitals in Nepal. The collection of demographic information was performed along with an entomological survey and clinical evaluation of the patients. A total of 280 serum samples were collected from suspected dengue patients. These samples were subjected to routine laboratory investigations and IgM-capture ELISA for dengue serological identification, and 160 acute serum samples were used for virus isolation, RT-PCR, sequencing and phylogenetic analysis. The results showed that affected patients were predominately adults, and that 10% of the cases were classified as dengue haemorrhagic fever/ dengue shock syndrome. The genetic characterization of dengue viruses isolated from patients in four major outbreak areas of Nepal suggests that the DENV-1 strain was responsible for the 2010 epidemic. Entomological studies identified Aedes aegypti in all epidemic areas. All viruses belonged to a monophyletic single clade which is phylogenetically close to Indian viruses. The dengue epidemic started in the lowlands and expanded to the highland areas. To our knowledge, this is the first dengue isolation and genetic characterization reported from Nepal.