Suman Rijal

B.P. Koirala Institute of Health Sciences

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Marleen Boelaert

Instituut voor Tropische Geneeskunde

Jean‐Claude Dujardin

Instituut voor Tropische Geneeskunde

François Chappuis

Hôpital Beau-Séjour

Basudha Khanal

B.P. Koirala Institute of Health Sciences

Shyam Sundar

Banaras Hindu University

Narayan Raj Bhattarai

B.P. Koirala Institute of Health Sciences

Albert Picado

Foundation for Innovative New Diagnostics

Bart Ostyn

Instituut voor Tropische Geneeskunde

Murari Lal Das

Rajendra Memorial Research Institute of Medical Sciences

Manu Vanaerschot

Columbia University Irving Medical Center

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Instituut voor Tropische Geneeskunde

114

B.P. Koirala Institute of Health Sciences

102

London School of Hygiene & Tropical Medicine

University of Antwerp

Banaras Hindu University

Research Institute for Tropical Medicine

Institute of Medical Sciences

Drugs for Neglected Diseases Initiative

World Health Organization

University of London

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Genomes of Leishmania parasites directly sequenced from patients with visceral leishmaniasis in the Indian subcontinent

PLoS neglected tropical diseases (2019)

Malgorzata A. Domagalska Hideo Imamura Mandy Sanders Frederik Van den Broeck Narayan Raj Bhattarai

Whole genome sequencing (WGS) is increasingly used for molecular diagnosis and epidemiology of infectious diseases. Current Leishmania genomic studies rely on DNA extracted from cultured parasites, which might introduce sampling and biological biases into the subsequent analyses. Up to now, direct analysis of Leishmania genome in clinical samples is hampered by high levels of human DNA and large variation in parasite load in clinical samples. Here, we present a method, based on target enrichment of Leishmania donovani DNA with Agilent SureSelect technology, that allows the analysis of Leishmania genomes directly in clinical samples. We validated our protocol with a set of artificially mixed samples, followed by the analysis of 63 clinical samples (bone marrow or spleen aspirates) from visceral leishmaniasis patients in Nepal. We were able to identify genotypes using a set of diagnostic SNPs in almost all of these samples (97%) and access comprehensive genome-wide information in most (83%). This allowed us to perform phylogenomic analysis, assess chromosome copy number and identify large copy number variants (CNVs). Pairwise comparisons between the parasite genomes in clinical samples and derived in vitro cultured promastigotes showed a lower aneuploidy in amastigotes as well as genomic differences, suggesting polyclonal infections in patients. Altogether our results underline the need for sequencing parasite genomes directly in the host samples

Amastigote

genomic DNA

10.1371/journal.pntd.0007900

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Citations (60)

<I>Phlebotomus argentipes</I> Seasonal Patterns in India and Nepal

Journal of Medical Entomology (2010)

Albert Picado Murari Lal Das Vijay Kumar Diwakar Singh Dinesh Suman Rijal

The current control of Phebotomus argentipes (Annandale and Brunetti), the vector of Leishmania donovani (Laveran and Mesnil), on the Indian subcontinent is base on indoor residual spraying. The efficacy of this method depends, among other factors, on the timing and number of spraying rounds, which depend on the P. argentipes seasonality. To describe P. argentipes' seasonal patterns, six visceral leishmaniasis (VL) endemic villages, three in Muzaffarpur and three in Sunsari districts in India and Nepal, respectively, were selected based on accessibility and VL incidence. Ten houses per cluster with the highest P. argentipes density were monitored monthly for 15-16 mo using Center for Disease Control and Prevention light traps. Minimum and maximum temperature and rainfall data for the months January 2006 through December 2007 were collected from the nearest available weather stations. Backwards stepwise regression was used to generate the minimal adequate model for explaining the monthly variation in P. argentipes populations. The seasonality of P. argentipes is similar in India and Nepal, with two annual density peaks around May and October. Monthly P. argentipes density is positively associated with temperature and negatively associated with rainfall in both study sites. The multivariate climate model explained 57% of the monthly vectorial abundance. Vector control programs against P. argentipes (i.e., indoor residual spraying) should take into account the seasonal described here when implementing and monitoring interventions. Monitoring simple meteorological variables (i.e., temperature, rainfall) may allow prediction of VL epidemics on the Indian subcontinent.

Indoor residual spraying

Seasonality

Indian subcontinent

10.1603/me09175

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Citations (39)

Lindane and Cetrimide lotion poisoning in an adult patient: A case report on an uncommon ingestion

Clinical Case Reports (2024)

Pragya Rai Shrijan Shrestha Suman Rijal Rakesh Singh

Key Clinical Message Lindane induces severe side effects, including fatality, while Cetrimide causes esophageal damage. With no antidotes available, our patient ingested both, requiring prompt gastric lavage and comprehensive treatment.

Cetrimide

Gastric lavage

Lindane

Lotion

10.1002/ccr3.8864

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Citations (1)

Adherence to miltefosine treatment for visceral leishmaniasis under routine conditions in Nepal

Tropical Medicine & International Health (2012)

Surendra Uranw Bart Ostyn Thomas P. C. Dorlo Epco Hasker Bruno Dujardin

Abstract Objective To assess patient adherence to unsupervised single‐drug miltefosine treatment for visceral leishmaniasis and to identify the factors influencing adherence. Methods This is a prospective cohort study of 171 patients with V isceral leishmaniasis ( VL ) in three healthcare settings in N epal. Adherence was assessed through pill count, checking of treatment cards and adherence questionnaires, as well as miltefosine concentration measurements at the end of treatment. Poor adherence was defined as less than 90% of required capsules taken. Results Patient adherence to miltefosine was 83%. Predictors of adherence were being the male sex ( OR = 2.60, 95% CI 1.02–6.67) and knowing the duration of treatment ( OR = 3.05, 95% CI 1.16–8.04). Adherence was also better for patients who were literate and knew the side effects of treatment. Gastrointestinal side effects and negligence after the resolution of clinical symptoms of VL were the main reasons for poor adherence. Poor adherence was associated (though not statistically significant) with future relapse. Conclusion Effective counselling during the treatment, a short take‐home message on VL and on side effects and action of miltefosine, and follow‐up visits are the best way to prevent poor adherence. Single end‐of‐treatment measurements of miltefosine concentrations as objective assessment of adherence would only be useful in addition to the subjective assessments when substantial doses of miltefosine have been missed.

Miltefosine

Pill

10.1111/tmi.12025

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Citations (19)

Epistaxis in Visceral Leishmaniasis with Hematological Correlation

International Journal of Otolaryngology (2012)

Brihaspati Sigdel Shital Bhandary Suman Rijal

Objective. To study the prevalence of epistaxis in visceral leismaniasis and its correlation with hematological profile. Methods. Out of 80 diagnosed cases of visceral leishmaniasis, 19 patients with epistaxis were included in the study. Diagnosis was made by Rk-39 from peripheral smear and LD bodies from bone marrow. Before starting anti-kala-azar treatment, nasal examination findings and hematological profile were noted. Study Design. Prospective cross-sectional hospital-based study. Results. Epistaxis was found in the age group of 7-66 years. Epistaxis was observed in 19 (23.8%) cases. One patient died because of epistaxis and neck hematoma. Conclusion. Epistaxis is a common ENT finding in endemic area of visceral leishmaniasis like our case.

10.1155/2012/809056

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Citations (4)

Importance of secondary screening with clinical isolates for anti-leishmania drug discovery

Scientific Reports (2018)

Aya Hefnawy Juan Cantizani Imanol Peña Pilar Manzano Suman Rijal

The growing drug resistance (DR) raises major concerns for the control of visceral leishmaniasis (VL), a neglected disease lethal in 95 percent of the cases if left untreated. Resistance has rendered antimonials (SSG) obsolete in the Indian Sub-Continent (ISC) and the first miltefosine-resistant Leishmania donovani were isolated. New chemotherapeutic options are needed and novel compounds are being identified by high-throughput screening (HTS). HTS is generally performed with old laboratory strains such as LdBOB and we aimed here to validate the activity of selected compounds against recent clinical isolates. In this academic/industrial collaboration, 130 compounds from the GSK "Leishbox" were screened against one SSG-sensitive and one SSG-resistant strain of L. donovani recently isolated from ISC patients, using an intracellular assay of L. donovani-infected THP1-derived macrophages. We showed that only 45% of the compounds were active in both clinical isolates and LdBOB. There were also different compound efficiencies linked to the SSG susceptibility background of the strains. In addition, our results suggested that the differential susceptibility profiles were chemical series-dependent. In conclusion, we demonstrate the potential value of including clinical isolates (as well as resistant strains) in the HTS progression cascade.

Miltefosine

10.1038/s41598-018-30040-5

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Citations (27)

Gene Expression Analysis of the Mechanism of Natural Sb(V) Resistance in Leishmania donovani Isolates from Nepal

Antimicrobial Agents and Chemotherapy (2005)

Saskia Decuypere Suman Rijal Vanessa Yardley Simonne De Doncker Thierry Laurent

Control of visceral leishmaniasis (VL) is being challenged by the emergence of natural resistance against the first line of treatment, pentavalent antimonials [Sb(V)]. An insight into the mechanism of natural Sb(V) resistance is required for the development of efficient strategies to monitor the emergence and spreading of Sb(V) resistance in countries where VL is endemic. In this work, we have focused on the mechanism of natural Sb(V) resistance emerging in Nepal, a site where anthroponotic VL is endemic. Based on the current knowledge of Sb(V) metabolism and of the in vitro trivalent antimonial [Sb(III)] models of resistance to Leishmania spp., we selected nine genes for a comparative transcriptomic study on natural Sb(V)-resistant and -sensitive Leishmania donovani isolates. Differential gene expression patterns were observed for the genes coding for 2-thiol biosynthetic enzymes, gamma-glutamylcysteine synthetase (GCS) and ornithine decarboxylase (ODC), and for the Sb(III) transport protein aquaglyceroporin 1 (AQP1). The results indicate that the mechanism for natural Sb(V) resistance partially differs from the mechanism reported for in vitro Sb(III) resistance. More specifically, we hypothesize that natural Sb(V) resistance results from (i) a changed thiol metabolism, possibly resulting in inhibition of Sb(V) activation in amastigotes, and (ii) decreased uptake of the active drug Sb(III) by amastigotes.

Amastigote

Miltefosine

10.1128/aac.49.11.4616-4621.2005

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Citations (147)

Evaluation of rapid diagnostic tests: visceral leishmaniasis

Nature Reviews Microbiology (2007)

Marleen Boelaert Sujit Bhattacharya François Chappuis Sayda Hassan El Safi Asrat Hailu

10.1038/nrmicro1766

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Citations (110)

Model-Based Investigations of Different Vector-Related Intervention Strategies to Eliminate Visceral Leishmaniasis on the Indian Subcontinent

PLoS neglected tropical diseases (2014)

Anette Stauch Hans-Peter Duerr Albert Picado Bart Ostyn Shyam Sundar

The elimination of infectious diseases requires reducing transmission below a certain threshold. The Visceral Leishmaniasis (VL) Elimination Initiative in Southeast Asia aims to reduce the annual VL incidence rate below 1 case per 10,000 inhabitants in endemic areas by 2015 via a combination of case management and vector control. Using a previously developed VL transmission model, we investigated transmission thresholds dependent on measures reducing the sand fly density either by killing sand flies (e.g., indoor residual spraying and long-lasting insecticidal nets) or by destroying breeding sites (e.g., environmental management). Model simulations suggest that elimination of VL is possible if the sand fly density can be reduced by 67% through killing sand flies, or if the number of breeding sites can be reduced by more than 79% through measures of environmental management. These results were compared to data from two recent cluster randomised controlled trials conducted in India, Nepal and Bangladesh showing a 72% reduction in sand fly density after indoor residual spraying, a 44% and 25% reduction through the use of long-lasting insecticidal nets and a 42% reduction after environmental management. Based on model predictions, we identified the parameters within the transmission cycle of VL that predominantly determine the prospects of intervention success. We suggest further research to refine model-based predictions into the elimination of VL.

Indoor residual spraying

Indian subcontinent

10.1371/journal.pntd.0002810

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Citations (39)

The economic burden of visceral leishmaniasis for households in Nepal

Transactions of the Royal Society of Tropical Medicine and Hygiene (2006)

Suman Rijal S Koirala Patrick Van der Stuyft Marleen Boelaert

Visceral leishmaniasis (VL) affects persons from the lowest socioeconomic strata of the community, but its economic impact is not precisely known. An exploratory survey to document the economic costs of VL to households was conducted in an endemic focus in eastern Nepal. Data were collected from the 20 households in this cluster. Cases of VL over the last 3 years were elicited and information on direct and indirect costs incurred due to the disease as well as income of the households over the last year was estimated. It was reported that 15.0% (16/107) of the residents had suffered from VL and that almost all of the patients had preferred, in the first instance, to visit the private services or local faith healers instead of visiting the local public health facility. Average total costs incurred per episode of VL were above the median annual per capita income, and six of the seven affected households either had to sell part of their livestock or to take a loan to cover the costs. Direct costs consisted of 53% of the total cost, with 75% of this cost incurred before the patients actually received any treatment for VL. This study demonstrates how VL can lead to catastrophic expenditure for affected households.

Economic cost

Per capita income

10.1016/j.trstmh.2005.09.017

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Citations (76)