Seroprevalence and Clinical Features of Scrub Typhus among Febrile Patients Attending a Referral Hospital in Kathmandu, Nepal.

(1) Background: Scrub typhus (ST) is endemic to Nepal. It is often underdiagnosed and misdiagnosed due to non-specific clinical presentation coupled with limited microbiological facilities, leading to adverse clinical outcomes. This study aimed to assess the seroprevalence of scrub typhus in febrile patients attending Sukraraj Tropical and Infectious Disease Hospital (STIDH), Nepal, from August 2018 to April 2019. (2) Materials and Method: Blood/serum samples and clinical and demographic data of adult febrile patients (≥19 years) who attended or were referred to the hospital were collected after obtaining written informed consent from the participants excluding immunocompromised individuals. Collected blood/serum samples were subjected to hematological, biochemical, and serological tests. A serological test for scrub typhus was performed using the ImmuneMed scrub typhus rapid diagnostic test kit. Data generated were analyzed using SPSS software version 24.0. (3) Results: Amongst the 2070 febrile patients, 462 (22.3%) were seropositive to at least one etiological agent of febrile illnesses (scrub typhus: 253 cases, dengue: 101 cases, leptospirosis: 9, brucellosis: 52, malaria: 9 and kala-azar: 20 cases). Scrub typhus accounted for 12.2% (n = 253) of total febrile illnesses followed by dengue (4.9%, n = 101). Mixed seropositivity of scrub typhus with dengue, brucellosis, and typhoid was found in 12 (0.6%), 9 (0.4%), and 5 (0.2%) cases, respectively. Among 253 scrub typhus patients, 53.4% were female. Among the 154 patients, the most common symptoms were fever (100%), headache (79.2%), sweating (70.1%), breathing difficulty (51.3%), redness of the eye (43.5%), and pathognomonic eschar was observed in 9.1% patients. Fifty percent of scrub typhus patients had low platelet count and >30% of patients had an elevated level of liver enzymes (such as serum glutamic oxaloacetic transaminase (SGPT) and serum glutamic pyruvic transaminase (SGOT). (4) Conclusion: Scrub typhus is a considerable cause of febrile illness in Nepal. Females apparently have a higher chance of acquiring scrub typhus. ST presents nonspecific clinical presentation. The diagnostic dilemma of typhus patients can be minimized by the early monitoring of ST-associated symptoms. The country's health system needs to be strengthened for early outbreak detection, and immediate response actions against scrub typhus to control the future outbreak of ST.

Disappearance of Influenza during the COVID-19 Pandemic: Nepal's Experience.

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Serological markers and molecular analysis of hepatitis B infection in a tertiary care hospital at Kathmandu, Nepal.

AIM: To analyze the serology and molecular markers of the hepatitis B-infected patients from the tertiary care hospital at Kathmandu in Nepal. METHODS: A total of 399 blood samples of patients from Sukraraj Tropical and Infectious Disease Hospital, Teku, Kathmandu, were collected. Samples were tested for HBsAg, HBeAg, and IgM anti-HBc using ELISA method. The samples were further categorized as acute and chronic. The genotyping was performed by real-time polymerase chain reaction (real-time PCR) and further validated by sequencing. RESULTS: Out of 399 samples that were collected, 271 and 128 samples were acute and chronic cases respectively. Fifty-six samples were genotyped by qPCR, out of which 40 samples belonged to genotype D, 4 to C/D recombinant, 5 to genotype C, 3 to genotype B, and 4 were genotype A respectively. From these, 15 samples were used for sequencing of P (polymerase) gene and S (surface) genes. Thus, obtained sequences were used to construct neighbor-joining tree using Tamura-Nei model evolution and further validated by Bayesian analysis. A total of four sub-genotypes namely A1, C1, D1, and D5 were detected. CONCLUSION: Hepatitis B virus infection is a global health problem affecting about 257 million people worldwide. In Nepal, there are few reports on the molecular and phylogenetic analysis of this virus. In this study, we report the circulation of seropositive occult hepatitis as well as CD-recombinant genotype in Nepalese population.

Prevalence and Predictors of Anemia in HIV-Infected Persons in Nepal.

BACKGROUND: Anemia is the commonest hematological complications in HIV patients, and has a significant impact on quality of life, morbidity, and mortality. However, little is known about the epidemiology of anemia in this population in a Nepalese setting. Therefore, the present study aimed at assessing the prevalence of anemia in patients living with HIV and further to determine the independent predictors associated with it. METHODS: This cross-sectional study was conducted in patients diagnosed with HIV at Sukraraj Tropical and Infectious Disease Hospital, Teku, Kathmandu from November 2016 to August 2017. Anemia was considered a core variable, and covariates used for analysis were age, sex, CD4 count, antiretroviral therapy regimen, history of intravenous drug use, marital status, religion, geography, employment status, hypertension, and diabetes mellitus. Prevalence of anemia and its independent predictors were evaluated. Fisher's exact and χ 2 tests were performed to determine the significance of differences among categorical variables and t-tests for continuous variables. Binary logistic regression was modeled to assess predictors associated with anemia. RESULTS: Of the total 210 patients analyzed, median age was 37.50±10.57 years, and 110 (52.6%) were male. The estimated prevalence of anemia overall was 66.7% (95% CI 60.64%-73.35%): mild anemia 14.3% (95% CI 8.25%-19.74%), moderate anemia 40.5% (95% CI 31.88%-48.11%), and severe anemia 11.9% (95% CI 6.61%-17.30%). Prevalence of anemia increased significantly with decreasing CD4 count: 5.71%, 12.85%, and 48.09% among patients with CD4 counts >500, 200-499, and <200 cells/mm3, respectively (P=0.019). Severity of anemia was significantly associated with immunostatus (<200, 200-499, and >500; P=0.048). Female sex was significantly associated with increased odds of anemia (OR 2.27, P=0.007). CONCLUSION: The present study demonstrated a high rate of anemia in a substantial number of HIV individuals. Therefore, early detection and timely management of anemia, especially in females and those with decreased immunostatus, are crucial to prevent anemia progression and improve quality of life.

Understanding COVID-19 in Nepal.

The novel coronavirus COVID-19 (SARS-CoV-2) was first reported in 31 December 2019 in Wuhan City, China. The first case of COVID-19 was officially announced on 24 January, 2020, in Nepal. Nine COVID-19 cases have been reported in Nepal. We aim to describe our experiences of COVID-19 patients in Nepal. Keywords: COVID-19; experience; Nepal.

Carriage of Extended-Spectrum-ß-Lactamase- and AmpC-ß-Lactamase-Producing Enterobacteriaceae (ESBL-PE) in Healthy Community and Outpatient Department (OPD) Patients in Nepal.

BACKGROUND: Extended-spectrum ß-lactamase (ESBL)- and AmpC-ß-lactamase (ESBL)- and AmpC-Enterobacteriaceae have recently emerged as a public threat in the treatment of nosocomial as well as community-acquired infections. Very little information is currently available about its existence in Nepal. We, therefore, aim to determine the prevalence of ESBL and AmpC-ß-lactamase (ESBL)- and AmpC-Enterobacteriaceae have recently emerged as a public threat in the treatment of nosocomial as well as community-acquired infections. Very little information is currently available about its existence in Nepal. We, therefore, aim to determine the prevalence of ESBL and AmpC. METHODS: During a 6-month period (November 2014-April 2015), a total of 190 stool specimens from 190 participants were obtained from different population. Of the total 260 fecal isolates, 152 from outpatient department (OPD) and 108 from healthy volunteer were collected. Stool specimens were cultured and enterobacterial isolates were subjected to antimicrobial susceptibility tests according to the standard microbiologic guidelines. ESBL was screened using ceftazidime (CAZ, 30 µg) and cefotaxime (CTX, 30 µg) and cefotaxime (CTX, 30 ß-lactamase (ESBL)- and AmpC. RESULTS: The prevalence of ESBL, AmpC-ß-lactamase (ESBL)- and AmpC-ß-lactamase (ESBL)- and AmpC-Enterobacteriaceae have recently emerged as a public threat in the treatment of nosocomial as well as community-acquired infections. Very little information is currently available about its existence in Nepal. We, therefore, aim to determine the prevalence of ESBL and AmpC-E. coli was 70.2% followed by K. pneumoniae (12.7%), and among AmpC-ß-lactamase (ESBL)- and AmpC-E. coli was 70.2% followed by E. coli was 70.2% followed by K. pneumoniae (12.7%), and among AmpC-K. pneumoniae (12.7%), and among AmpC-C. freundii 2/7 (28.57%) were detected highest among AmpC-ß-lactamase (ESBL)- and AmpC. CONCLUSION: Our study revealed a high prevalence of ESBL- and AmpC-ß-lactamase-producing enteric pathogen in Nepalese OPD and healthy population. The significant increase of these isolates and increased rate of drug resistance indicates a serious threat that stress the need to implement the surveillance system and a proper control measure so as to limit the spread of ESBL-producing Enterobacteriaceae (ESBL-PE) in both OPD as well as in community. Therefore, healthcare providers need to be aware that ESBL- and AmpC-ß-lactamase-producing strains are not only circulating in hospital environments but also in the community and should be dealt with accordingly.ß-lactamase (ESBL)- and AmpC-Enterobacteriaceae have recently emerged as a public threat in the treatment of nosocomial as well as community-acquired infections. Very little information is currently available about its existence in Nepal. We, therefore, aim to determine the prevalence of ESBL and AmpC-ß-lactamase (ESBL)- and AmpC.

Burden of Shigella spp and Vibrio spp, and their antibiotic sensitivity pattern in the patients with acute gastroenteritis in tertiary care hospital in Nepal.

OBJECTIVES: The present study aims to investigate the etiology, clinical profile and resistance pattern of the isolated pathogens in Nepalese adults with acute gastroenteritis. This cross-sectional study was conducted at Sukraraj Tropical and Infectious Disease Hospital, from April 2016 to Sep 2017. Subjects' ages 14 or above, presenting with gastroenteritis with positive stool culture were enrolled for analysis. RESULTS: Of total 153 patients, 47.72% subjects confirmed the presence of bacterial infection. Vibrio cholerae spp and Shigella spp were detected in 36.6% and 23.28% respectively. The most common resistance among Vibrio cholerae was to nitrofurantoin (92.8%), cotrimoxazole (92.8%) and nalidixic acid (92.8%). Among 17 isolates of Shigella spp, the most frequent drug resistant was observed in ampicillin (64.7%), nalidixic acid (58.8%), ceftriaxone (47%). Chloramphenicol (94.1%), tetracycline (88.2%), and cotrimoxazole (82.3%) were found to be the most sensitive towards this pathogen. High rate of diarrhea due to bacterial infection, especially Shigella spp and Vibrio spp and their high rate of drug resistance emphasize an urgent need of designing a surveillance system for antimicrobial resistance in Nepalese setting.

Molecular evidence supports the expansion of visceral leishmaniasis towards non-program districts of Nepal.

BACKGROUND: Visceral Leishmaniasis (VL) is caused by a protozoan parasite Leishmania donovani that is transmitted to humans by an infected female sandfly, Phlebotomus argentipes. VL is common in the Indian sub-continent including Nepal and efforts for its elimination are ongoing. However, expansion of disease towards the higher altitude areas, previously considered as VL free in Nepal, may impact the ability to achieve the elimination target by 2020. METHODS: This was an exploratory study, where VL suspected patients living exclusively in the non-program districts of Nepal and presenting with fever > 2 weeks and splenomegaly was included. The patients' blood samples were collected, and DNA was extracted. DNA was subjected to PCR amplification and subsequent sequencing. Additionally, past 10 years data of VL cases from the national databases were analysed to see the trends of the disease in program and non program districts. RESULTS: Analysis of the past 10 years data revealed that trend of VL cases significantly decreased in the program districts (p = 0.001) while it increased in the non-program districts (p = 0.002). The national trend for overall incidence of VL also significantly decreased over this time period. Limited number of patients' samples (n = 14) were subjected to molecular investigation, and four patients were found to be positive for Leishmania species by PCR. Interestingly, these cases in non-program districts were indeed also L. donovoni complex. All four patients were male with age ranges from 10 to 68 years. GenBank BLAST of the obtained DNA sequences confirmed identified specimens as L. donovani complex. We identified additional VL cases from non-program districts (including the high lands) of Nepal, indicating that the infection could be an emerging threat for the non-program areas of Nepal. CONCLUSION: The demonstration of VL cases in areas initially considered non-endemic has raised concern about on-going transmission in those regions and may trigger subsequent government plan and action to include those areas in the elimination program. Thus, the government should consider revising the disease control programs to accommodate non-program districts for achieving the VL elimination goal set for 2020.

Whole genome characterisation of G11P[25] and G9P[19] rotavirus A strains from adult patients with diarrhoea in Nepal.

Rotavirus A (RVA) causes acute diarrhoea in children and less frequently in adults. However, the knowledge about the genotype distribution of RVA strains circulating in adults is limited particularly in developing countries. This study aimed to characterise the RVA strains detected from adult patients with diarrhoea in Nepal. A total of 47 RVA positive stool samples from adult patients with diarrhoea in Kathmandu, Nepal during 2007-2008 were examined for the G and P genotypes by sequencing. Nearly half (49%) of the samples were genotyped as G9P[8] (n = 23), G1P[8], G2P[4] (n = 5 each), G12P[8] (n = 4), G12P[6] (n = 3), G1P[6] (n = 2), G3P[8] and G9P[6] (n = 1 each). Interestingly, two G11P[25] and one G9P[19] strains detected were further subjected to Illumina MiSeq next generation sequencing to determine their whole genome sequences. The genotype constellations of RVA/Human-wt/NPL/TK2615/2008/G11P[25] and RVA/Human-wt/NPL/TK2620/2008/G11P[25] were I12-R1-C1-M1-A1-N1-T1-E1-H1, whereas that of RVA/Human-wt/NPL/TK1797/2007/G9P[19] was I5-R1-C1-M1-A8-N1-T1-E1-H1. The 11 genes of TK2615 and TK2620 were virtually identical, and they were either porcine-like or unique except the VP2 and NSP1 genes which were of human RVA origin. The two G11P[25] strains were also very similar to KTM368, another G11P[25] isolated from a child in Nepal in 2004. On the other hand, no gene of TK1797 was likely to be of human RVA origin. The observation that porcine-like RVAs were detected from adult patients justifies further studies to explore the role of adults in the interspecies transmission of animal RVA to humans.

Visceral leishmaniasis from a non-endemic Himalayan region of Nepal.

Visceral leishmaniasis (VL) is endemic to the southern plains of Nepal. Here, we report the first case of VL from a non-endemic Himalayan region of Nepal. The patient presented with a history of high-grade fever, splenomegaly, and anemia but had not traveled to a VL-endemic region. Visceral leishmaniasis was diagnosed following microscopic detection of the Leishmania species amastigote in a bone marrow aspirate, positive result for the rK39 test, and further validation by nested polymerase chain reaction (PCR). The patient was treated with 5 mg/kg liposomal amphotericin B and was clinically improved upon discharge. Our result suggests that VL is expanding towards non-endemic regions of Nepal, and it should therefore be considered that VL surveillance systems be strengthened, particularly for non-program districts and VL be included as a differential diagnosis in febrile illnesses.

First report of Lyme disease in Nepal.

INTRODUCTION: Lyme disease is a tick-borne illness caused by the spirochete Borrelia burgdorferi and is widely reported in the USA, Central Europe, South East Asia and Latin America. Until recently, no scientific report regarding Lyme disease in Nepal had been published. CASE PRESENTATION: A 32-year-old, previously healthy female visited the hospital with a history of joint pains, fatigue, neck stiffness, tingling sensation and headache. She was initially treated for typhoid fever, brucellosis and malaria, but did not show significant improvement. Doxycycline was prescribed empirically for 3 weeks for the treatment of suspected tick-borne illness. A two-tiered immunoglobulin laboratory testing confirmed Borrelia burgdorferi. She developed post-treatment Lyme disease syndrome after completion of antibiotic therapy. CONCLUSION: To the best of our knowledge, this is the first report of Lyme disease in Nepal and probably the first documented case of post-treatment Lyme disease syndrome in Asia. Lyme disease might have been overlooked in Nepal and, therefore, patients having clinical signs and symptoms similar to Lyme disease should not be disregarded in differential diagnosis.

Epidemiology, Impact and Control of Rabies in Nepal: A Systematic Review.

BACKGROUND: Rabies is a vaccine-preventable viral zoonosis belonging to the group of neglected tropical diseases. Exposure to a rabid animal may result in a fatal acute encephalitis if effective post-exposure prophylaxis is not provided. Rabies occurs worldwide, but its burden is disproportionately high in developing countries, including Nepal. We aimed to summarize current knowledge on the epidemiology, impact and control of rabies in Nepal. METHODS: We performed a systematic review of international and national scientific literature and searched grey literature through the World Health Organization Digital Library and the library of the National Zoonoses and Food Hygiene Research Centre, Nepal, and through searching Google and Google Scholar. Further data on animal and human rabies were obtained from the relevant Nepalese government agencies. Finally, we surveyed the archives of a Nepalese daily to obtain qualitative information on rabies in Nepal. FINDINGS: So far, only little original research has been conducted on the epidemiology and impact of rabies in Nepal. Per year, rabies is reported to kill about 100 livestock and 10-100 humans, while about 1,000 livestock and 35,000 humans are reported to receive rabies post-exposure prophylaxis. However, these estimates are very likely to be serious underestimations of the true rabies burden. Significant progress has been made in the production of cell culture-based anti-rabies vaccine and rabies immunoglobulin, but availability and supply remain a matter of concern, especially in remote areas. Different state and non-state actors have initiated rabies control activities over the years, but efforts typically remained focalized, of short duration and not harmonized. Communication and coordination between veterinary and human health authorities is limited at present, further complicating rabies control in Nepal. Important research gaps include the reporting biases for both human and animal rabies, the ecology of stray dog populations and the true contribution of the sylvatic cycle. INTERPRETATION: Better data are needed to unravel the true burden of animal and human rabies. More collaboration, both within the country and within the region, is needed to control rabies. To achieve these goals, high level political commitment is essential. We therefore propose to make rabies the model zoonosis for successful control in Nepal.

First report of Chikungunya virus infection in Nepal.

Chikungunya virus is an emerging arboviral disease that has been spreading rapidly across south Asia in recent years. Until recently, no chikungunya cases have been reported in Nepal. For the first time, we report three cases of chikungunya virus infection in Nepal.

The burden of parasitic zoonoses in Nepal: a systematic review.

BACKGROUND: Parasitic zoonoses (PZs) pose a significant but often neglected threat to public health, especially in developing countries. In order to obtain a better understanding of their health impact, summary measures of population health may be calculated, such as the Disability-Adjusted Life Year (DALY). However, the data required to calculate such measures are often not readily available for these diseases, which may lead to a vicious circle of under-recognition and under-funding. METHODOLOGY: We examined the burden of PZs in Nepal through a systematic review of online and offline data sources. PZs were classified qualitatively according to endemicity, and where possible a quantitative burden assessment was conducted in terms of the annual number of incident cases, deaths and DALYs. PRINCIPAL FINDINGS: Between 2000 and 2012, the highest annual burden was imposed by neurocysticercosis and congenital toxoplasmosis (14,268 DALYs [95% Credibility Interval (CrI): 5450-27,694] and 9255 DALYs [95% CrI: 6135-13,292], respectively), followed by cystic echinococcosis (251 DALYs [95% CrI: 105-458]). Nepal is probably endemic for trichinellosis, toxocarosis, diphyllobothriosis, foodborne trematodosis, taeniosis, and zoonotic intestinal helminthic and protozoal infections, but insufficient data were available to quantify their health impact. Sporadic cases of alveolar echinococcosis, angiostrongylosis, capillariosis, dirofilariosis, gnathostomosis, sparganosis and cutaneous leishmaniosis may occur. CONCLUSIONS/SIGNIFICANCE: In settings with limited surveillance capacity, it is possible to quantify the health impact of PZs and other neglected diseases, thereby interrupting the vicious circle of neglect. In Nepal, we found that several PZs are endemic and are imposing a significant burden to public health, higher than that of malaria, and comparable to that of HIV/AIDS. However, several critical data gaps remain. Enhanced surveillance for the endemic PZs identified in this study would enable additional burden estimates, and a more complete picture of the impact of these diseases.

Critical phase among patients with dengue fever during the 2010 outbreak in Nepal.

BACKGROUND: Dengue virus infection has become an increasingly serious threat to public health in Nepal. Detection and/or serotyping of dengue virus has been reported in the past; however, little is known about the critical phase among dengue patients in Nepal. METHODS: Rapid immunochromatographic tests and reverse transcription PCR (RT-PCR) were performed in order to diagnosis dengue fever. Ultrasonographic and chest radiography were used to identify the plasma leakage phase. RESULTS: Of the total 264 patients with dengue, 8.7% (23/264) were admitted during the critical phase; 82.6% (19/23) of the patients were between 20 and 44 years of age. CONCLUSION: Physicians need to be vigilant and attentive to young adult patients with dengue fever.

A series of case reports of autochthonous visceral leishmaniasis, mostly in non-endemic hilly areas of Nepal.

The government of Nepal has committed to eliminate visceral leishmaniasis (VL) by 2015. The expansion of VL into new areas would constitute a major obstacle to achieving this goal. We report a series of autochthonous VL cases from areas currently considered non-endemic, mostly in hilly regions of Nepal.

Rotavirus, vaccine and unanswered questions: a perspective from a least developed country.

Two rotavirus vaccines, RotaTeq (Merck) and Rotarix (GlaxoSmithKline) have been developed to neutralize the most common rotavirus serotypes, and are now available in the global market. These vaccines are primarily aimed at reducing rotavirus gastroenteritis in children in the least developed countries, where rotavirus mortality rate is believed to be greatest. Thus, the World Health Organization (WHO) has recommended rotavirus vaccination be included in all national immunization programs, while the least developed countries have so far not come up with clear vision and long term strategy on vaccine implementation, and several questions, in addition to this, remain unanswered.