ABSTRACT
In the Anopheles genus, various mosquito species are able to transmit the Plasmodium parasites responsible for malaria, while others are non-vectors. In an effort to better understand the biology of Anopheles species and to quantify transmission risk in an area, the identification of mosquito species collected in the field is an essential but problematic task. Morphological identification requires expertise and cannot be checked after processing samples in a destructive treatment, while sequencing of numerous samples is costly. Here, we introduce a method of Species identification via Simple Observation Coupled with Capillary Electrophoresis Technology (SOCCET). This molecular technique of species identification is based on precise determination of ITS2 length combined with a simple visual observation, the colour of mosquito hindleg tip. DNA extracted from field-collected Anopheles mosquitoes was amplified with universal Anopheles ITS2 primers and analysed with a capillary electrophoresis device, which precisely determines the size of the fragments. We defined windows of amplicon sizes combined with fifth hind tarsus colour, which allows discrimination of the major Anopheles species found in our collections. We validated our parameters via Sanger sequencing of ITS2 amplicons. Using the SOCCET method, we characterised the composition of Anopheles populations in five locations of French Guiana, where we detected a total of nine species. Anopheles braziliensis and Anopheles darlingi were detected in four locations each and represented 13 and 67% of our samples, respectively. The SOCCET method can be particularly useful when working with routine sampling sites with a moderate species diversity, that is, when the number of local species is too high to define species-specific primers but low enough to avoid individual ITS2 sequencing. This tool will be of interest to evaluate local malaria transmission risk and this approach may be further implemented for other mosquito genera.
ABSTRACT
Cutaneous leishmaniasis (CL) caused by infection with the parasite Leishmania exhibits a large spectrum of clinical manifestations ranging from single healing to severe chronic lesions with the manifestation of resistance or not to treatment. Depending on the specie and multiple environmental parameters, the evolution of lesions is determined by a complex interaction between parasite factors and the early immune responses triggered, including innate and adaptive mechanisms. Moreover, lesion resolution requires parasite control as well as modulation of the pathologic local inflammation responses and the initiation of wound healing responses. Here, we have summarized recent advances in understanding the in situ immune response to cutaneous leishmaniasis: i) in North Africa caused by Leishmania (L.) major, L. tropica, and L. infantum, which caused in most cases localized autoresolutives forms, and ii) in French Guiana resulting from L. guyanensis and L. braziliensis, two of the most prevalent strains that may induce potentially mucosal forms of the disease. This review will allow a better understanding of local immune parameters, including cellular and cytokines release in the lesion, that controls infection and/or protect against the pathogenesis in new world compared to old world CL.
Subject(s)
Leishmania , Leishmaniasis, Cutaneous , Humans , French Guiana/epidemiology , Africa, Northern , CytokinesABSTRACT
Changes in biodiversity may impact infectious disease transmission through multiple mechanisms. We explored the impact of biodiversity changes on the transmission of Amazonian leishmaniases, a group of wild zoonoses transmitted by phlebotomine sand flies (Psychodidae), which represent an important health burden in a region where biodiversity is both rich and threatened. Using molecular analyses of sand fly pools and blood-fed dipterans, we characterized the disease system in forest sites in French Guiana undergoing different levels of human-induced disturbance. We show that the prevalence of Leishmania parasites in sand flies correlates positively with the relative abundance of mammal species known as Leishmania reservoirs. In addition, Leishmania reservoirs tend to dominate in less diverse mammal communities, in accordance with the dilution effect hypothesis. This results in a negative relationship between Leishmania prevalence and mammal diversity. On the other hand, higher mammal diversity is associated with higher sand fly density, possibly because more diverse mammal communities harbor higher biomass and more abundant feeding resources for sand flies, although more research is needed to identify the factors that shape sand fly communities. As a consequence of these antagonistic effects, decreased mammal diversity comes with an increase of parasite prevalence in sand flies, but has no detectable impact on the density of infected sand flies. These results represent additional evidence that biodiversity changes may simultaneously dilute and amplify vector-borne disease transmission through different mechanisms that need to be better understood before drawing generalities on the biodiversity-disease relationship.
Subject(s)
Leishmania , Leishmaniasis , Psychodidae , Animals , Humans , Leishmania/genetics , Biodiversity , Zoonoses , MammalsABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Leishmaniasis are widely distributed among tropical and subtropical countries, and remains a crucial health issue in Amazonia. Indigenous groups across Amazonia have developed abundant knowledge about medicinal plants related to this pathology. AIM OF THE STUDY: We intent to explore the weight of different pharmacological activities driving taxa selection for medicinal use in Amazonian communities. Our hypothesis is that specific activity against Leishmania parasites is only one factor along other (anti-inflammatory, wound healing, immunomodulating, antimicrobial) activities. MATERIALS AND METHODS: The twelve most widespread plant species used against leishmaniasis in Amazonia, according to their cultural and biogeographical importance determined through a wide bibliographical survey (475 use reports), were selected for this study. Plant extracts were prepared to mimic their traditional preparations. Antiparasitic activity was evaluated against promastigotes of reference and clinical New-World strains of Leishmania (L. guyanensis, L. braziliensis and L. amazonensis) and L. amazonensis intracellular amastigotes. We concurrently assessed the extracts immunomodulatory properties on PHA-stimulated human PBMCs and RAW264.7 cells, and on L. guyanensis antigens-stimulated PBMCs obtained from Leishmania-infected patients, as well as antifungal activity and wound healing properties (human keratinocyte migration assay) of the selected extracts. The cytotoxicity of the extracts against various cell lines (HFF1, THP-1, HepG2, PBMCs, RAW264.7 and HaCaT cells) was also considered. The biological activity pattern of the extracts was represented through PCA analysis, and a correlation matrix was calculated. RESULTS: Spondias mombin L. bark and Anacardium occidentale L. stem and leaves extracts displayed high anti-promatigotes activity, with IC50 ≤ 32 µg/mL against L. guyanensis promastigotes for S. mombin and IC50 of 67 and 47 µg/mL against L. braziliensis and L. guyanensis promastigotes, respectively, for A. occidentale. In addition to the antiparasitic effect, antifungal activity measured against C. albicans and T. rubrum (MIC in the 16-64 µg/mL range) was observed. However, in the case of Leishmania amastigotes, the most active species were Bixa orellana L. (seeds), Chelonantus alatus (Aubl.) Pulle (leaves), Jacaranda copaia (Aubl.) D. Don. (leaves) and Plantago major L. (leaves) with IC50 < 20 µg/mL and infection rates of 14-25% compared to the control. Concerning immunomodulatory activity, P. major and B. orellana were highlighted as the most potent species for the wider range of cytokines in all tested conditions despite overall contrasting results depending on the model. Most of the species led to moderate to low cytotoxic extracts except for C. alatus, which exhibited strong cytotoxic activity in almost all models. None of the tested extracts displayed wound healing properties. CONCLUSIONS: We highlighted pharmacologically active extracts either on the parasite or on associated pathophysiological aspects, thus supporting the hypothesis that antiparasitic activities are not the only biological factor useful for antileishmanial evaluation. This result should however be supplemented by in vivo studies, and attracts once again the attention on the importance of the choice of biological models for an ethnophamacologically consistent study. Moreover, plant cultural importance, ecological status and availability were discussed in relation with biological results, thus contributing to link ethnobotany, medical anthropology and biology.
Subject(s)
Antiprotozoal Agents/pharmacology , Leishmania/drug effects , Plant Extracts/pharmacology , Plants, Medicinal/chemistry , Animals , Antiprotozoal Agents/isolation & purification , Brazil , HaCaT Cells , Hep G2 Cells , Humans , Leishmaniasis/drug therapy , Leishmaniasis/parasitology , Leukocytes, Mononuclear/parasitology , Medicine, Traditional , Mice , RAW 264.7 Cells , THP-1 CellsABSTRACT
BACKGROUND: Cutaneous Leishmaniasis (CL) is endemic in French Guiana but cases are usually sporadic. An outbreak signal was issued on May 15th 2020 with 15 suspected cases after a military training course in the rainforest. An outbreak investigation was carried out. METHODOLOGY/PRINCIPAL FINDINGS: Thirty cases were confirmed. Leishmania guyanensis was the most frequent species (90%). The most frequent presentation was ulcerative (90%). Lesions on the face and hands were frequent (40% each). Eight cases (26%) presented a poor outcome after treatment with pentamidine and required a second line with amphotericin B. Three of them required further treatments with meglumine antimoniate or miltefosine. Two spots within the training area were deemed as likely sites of contamination, due to illegal logging. The isolated Leishmania strains did not form a separate cluster. Participation in Week 13 of year 2020 was associated with infection (OR = 4.59 [1.10-19.83]; p = 0.016) while undergoing only the "Fighting" exercise was protective (OR = 0.1 [0-0.74]; p = 0.021). There was no association between infection and other risk factors at the individual level. The attack rate of Regiment B (14/105 = 13.3%) was significantly higher (OR = 4.22 [1.84-9.53], p = 0.0001) compared to Regiment A (16/507 = 3.2%). The attack rate during this training course (30/858 = 3.5%) was significantly higher (OR 2.29 [1.28-4.13]; p = 0.002) than for other missions in French Guiana during the same period (22/1427 = 1.5%). CONCLUSIONS: This outbreak could be explained by a combination of factors: climatic conditions around week 13, at-risk activities including night trainings, absence of impregnation, a lesser experience of rainforest duties in Regiment B and illegal logging attracting sandflies on military training grounds.
Subject(s)
Leishmania/isolation & purification , Leishmaniasis, Cutaneous/epidemiology , Leishmaniasis, Cutaneous/parasitology , Military Personnel/statistics & numerical data , Phylogeny , Adult , Antiprotozoal Agents/administration & dosage , Disease Outbreaks , Female , French Guiana/epidemiology , Humans , Leishmania/drug effects , Leishmania/genetics , Leishmania/physiology , Leishmaniasis, Cutaneous/diagnosis , Leishmaniasis, Cutaneous/drug therapy , Male , Middle Aged , Pentamidine/administration & dosage , Young AdultABSTRACT
Cutaneous Leishmaniasis (CL) is the most prevalent form of Leishmaniasis and is widely endemic in the Americas. Several species of Leishmania are responsible for CL, a severely neglected tropical disease and the treatment of CL vary according to the different species of Leishmania. We proposed to map the distribution of the Leishmania species reported in French Guiana (FG) using a biogeographic approach based on environmental predictors. We also measured species endemism i.e., the uniqueness of species to a defined geographic location. Our results show that the distribution patterns varied between Leishmania spp. and were spatially dependent on climatic covariates. The species distribution modelling of the eco-epidemiological spatial patterns of Leishmania spp. is the first to measure endemism based on bioclimatic factors in FG. The study also emphasizes the impact of tree cover loss and climate on the increasing distribution of L. (Viannia) braziliensis in the most anthropized regions. Detection of high-risk regions for the different between Leishmania spp. is essential for monitoring and active surveillance of the vector. As climate plays a major role in the spatial distribution of the vector and reservoir and the survival of the pathogen, climatic covariates should be included in the analysis and mapping of vector-borne diseases. This study underscores the significance of local land management and the urgency of considering the impact of climate change in the development of vector-borne disease management strategies at the global scale.
ABSTRACT
French Guiana is a European ultraperipheric region located on the northern Atlantic coast of South America. It constitutes an important forested region for biological conservation in the Neotropics. Although very sparsely populated, with its inhabitants mainly concentrated on the Atlantic coastal strip and along the two main rivers, it is marked by the presence and development of old and new epidemic disease outbreaks, both research and health priorities. In this review paper, we synthetize 15 years of multidisciplinary and integrative research at the interface between wildlife, ecosystem modification, human activities and sociodemographic development, and human health. This study reveals a complex epidemiological landscape marked by important transitional changes, facilitated by increased interconnections between wildlife, land-use change and human occupation and activity, human and trade transportation, demography with substantial immigration, and identified vector and parasite pharmacological resistance. Among other French Guianese characteristics, we demonstrate herein the existence of more complex multi-host disease life cycles than previously described for several disease systems in Central and South America, which clearly indicates that today the greater promiscuity between wildlife and humans due to demographic and economic pressures may offer novel settings for microbes and their hosts to circulate and spread. French Guiana is a microcosm that crystallizes all the current global environmental, demographic and socioeconomic change conditions, which may favor the development of ancient and future infectious diseases.
Subject(s)
Animals, Wild , Demography , Ecosystem , Vector Borne Diseases , Zoonoses , Animals , French Guiana/epidemiology , Human Activities , Humans , Incidence , Interdisciplinary Research , Prevalence , Vector Borne Diseases/epidemiology , Vector Borne Diseases/transmission , Zoonoses/epidemiology , Zoonoses/etiology , Zoonoses/transmissionABSTRACT
Recent studies have highlighted the interest in noninvasive sampling procedures coupled with real-time PCR methods for the detection of Leishmania species in South America. In French Guiana, the sampling method still relied on skin biopsies. Noninvasive protocols should be tested on a large annual cohort to improve routine laboratory diagnosis of cutaneous leishmaniasis. Therefore, we evaluated the performance of a new Leishmania detection and species identification protocol involving cotton swabs and SYBR green-based real-time PCR of the Hsp70 gene, coupled with Sanger sequencing. Between May 2017 and May 2018, 145 patients with ulcerated lesions compatible with cutaneous leishmaniasis were included in the study at the Cayenne Hospital and its remote health centers. Each patient underwent scrapings for a smear, skin biopsies for parasite culture and PCR-restriction fragment length polymorphism (RFLP) (RNA polymerase II), and sampling with a cotton swab for SYBR green-based PCR. The most accurate diagnostic test was the SYBR green-based PCR on swab samples, showing 98% sensitivity. The mean PCR cycle threshold (CT ) was 24.4 (minimum CT , 17; maximum CT , 36) and was <35 in 97.6% of samples. All samples positive by SYBR green-based real-time PCR were successfully identified at the species level by DNA sequencing. This new method should be considered for routine diagnosis of cutaneous leishmaniasis in South America and especially for remote areas, since noninvasive collection tools are easier to use and require fewer precautions for transportation.
Subject(s)
Leishmaniasis, Cutaneous , DNA, Protozoan/genetics , French Guiana , Humans , Leishmaniasis, Cutaneous/diagnosis , Polymorphism, Restriction Fragment Length , Real-Time Polymerase Chain Reaction , Sensitivity and Specificity , South AmericaABSTRACT
The mosquito microbiota reduces the vector competence of Anopheles to Plasmodium and affects host fitness; it is therefore considered as a potential target to reduce malaria transmission. While immune induction, secretion of antimicrobials and metabolic competition are three typical mechanisms of microbiota-mediated protection against invasive pathogens in mammals, the involvement of metabolic competition or mutualism in mosquito-microbiota and microbiota-Plasmodium interactions has not been investigated. Here, we describe a metabolome analysis of the midgut of Anopheles coluzzii provided with a sugar-meal or a non-infectious blood-meal, under conventional or antibiotic-treated conditions. We observed that the antibiotic treatment affects the tricarboxylic acid cycle and nitrogen metabolism, notably resulting in decreased abundance of free amino acids. Linking our results with published data, we identified pathways which may participate in microbiota-Plasmodium interactions via metabolic interactions or immune modulation and thus would be interesting candidates for future functional studies.
ABSTRACT
In French Guiana, five species are associated with Cutaneous Leishmaniasis (CL). Though infections with Leishmania guyanensis, L. (V.) braziliensis and L. (L.) amazonensis have been extensively described, there are few available clinical and genetic data on L. (V.) lainsoni and L. (V.) naiffi. We determined the clinical and epidemiological features of all cases of CL due to L. (V.) naiffi and L. (V.) lainsoni diagnosed in French Guiana between 2003 and 2019. Phylogenetic analysis was performed by sequencing a portion of HSP70 and cyt b genes. Five cases of L. naiffi and 25 cases of L. lainsoni were reported. Patients infected by L. (V.) lainsoni were usually infected on gold camps, mostly along the Maroni river (60%), while L. naiffi was observed in French patients infected on the coast (100%). A high number of pediatric cases (n = 5; 20%) was observed for L. (V.) lainsoni. A mild clinical course was observed for all cases of L. (V.) naiffi. HSP70 and cyt b partial nucleotide sequence analysis revealed different geographical clusters within L. (V.) naiffi and L. (V.) lainsoni but no association were found between phylogenetic and clinical features. Our data suggest distinct socio-epidemiological features for these two Leishmania species. Patients seem to get infected with L. (V.) naiffi during leisure activities in anthropized coastal areas, while L. (V.) lainsoni shares common features with L. (V.) guyanensis and braziliensis and seems to be acquired during professional activities in primary forest regions. Phylogenetic analysis has provided information on the intraspecific genetic variability of L. (V.) naiffi and L. (V.) lainsoni and how these genotypes are distributed at the geographic level.
Subject(s)
Leishmania/classification , Leishmaniasis, Cutaneous/drug therapy , Leishmaniasis, Cutaneous/epidemiology , Adolescent , Adult , Child , Child, Preschool , Cytochromes b/genetics , Female , French Guiana/epidemiology , HSP70 Heat-Shock Proteins/genetics , Humans , Infant , Leishmania/genetics , Leishmania/pathogenicity , Leishmaniasis, Cutaneous/pathology , Leisure Activities , Male , Middle Aged , Mining , Neglected Diseases , Phylogeny , Retrospective Studies , Sequence Analysis, DNAABSTRACT
A major challenge of eco-epidemiology is to determine which factors promote the transmission of infectious diseases and to establish risk maps that can be used by public health authorities. The geographic predictions resulting from ecological niche modelling have been widely used for modelling the future dispersion of vectors based on the occurrence records and the potential prevalence of the disease. The establishment of risk maps for disease systems with complex cycles such as cutaneous leishmaniasis (CL) can be very challenging due to the many inference networks between large sets of host and vector species, with considerable heterogeneity in disease patterns in space and time. One novelty in the present study is the use of human CL cases to predict the risk of leishmaniasis occurrence in response to anthropogenic, climatic and environmental factors at two different scales, in the Neotropical moist forest biome (Amazonian basin and surrounding forest ecosystems) and in the surrounding region of French Guiana. With a consistent data set never used before and a conceptual and methodological framework for interpreting data cases, we obtained risk maps with high statistical support. The predominantly identified human CL risk areas are those where the human impact on the environment is significant, associated with less contributory climatic and ecological factors. For both models this study highlights the importance of considering the anthropogenic drivers for disease risk assessment in human, although CL is mainly linked to the sylvatic and peri-urban cycle in Meso and South America.
Subject(s)
Ecology , Ecosystem , Forests , Leishmaniasis, Cutaneous/epidemiology , French Guiana/epidemiology , Humans , Prevalence , Seasons , South America/epidemiologyABSTRACT
INTRODUCTION: American cutaneous leishmaniasis (ACL) is regarded as a public health problem in the Oiapoque basin, between Brazil and French Guiana. METHODS: Data on ACL occurrence/epidemiological profile and etiology were sourced from Brazilian health services and a reference laboratory. Rainfall correlation was also analyzed. RESULTS: Majority of the ACL cases were observed in adult men working as gold miners. ACL incidence peaks appeared to be linked to periods 2 months after the dry season. Migratory flow was found to be a non-negligible complicating factor in epidemiological surveillance. CONCLUSIONS: Binational strategies are required to minimize exposure for high-risk populations.
Subject(s)
Leishmaniasis, Cutaneous/epidemiology , Adult , Aged , Brazil/epidemiology , Female , French Guiana/epidemiology , Humans , Incidence , Male , Middle Aged , Seasons , Young AdultABSTRACT
Phlebotomine sand flies are insects that are highly relevant in medicine, particularly as the sole proven vectors of leishmaniasis. Accurate identification of sand fly species is an essential prerequisite for eco-epidemiological studies aiming to better understand the disease. Traditional morphological identification is painstaking and time-consuming, and molecular methods for extensive screening remain expensive. Recent studies have shown that matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a promising tool for rapid and cost-effective identification of arthropod vectors, including sand flies. The aim of this study was to validate the use of MALDI-TOF MS for the identification of Northern Amazonian sand flies. We constituted a MALDI-TOF MS reference database comprising 29 species of sand flies that were field-collected in French Guiana, which are expected to cover many of the more common species of the Northern Amazonian region, including known vectors of leishmaniasis. Carrying out a blind test, all the sand flies tested (n = 157) with a log (score) threshold greater than 1.7 were correctly identified at the species level. We confirmed that MALDI-TOF MS protein profiling is a useful tool for the study of sand flies, including neotropical species, known for their great diversity. An application that includes the spectra generated here will be available to the scientific community in the near future via an online platform.
Subject(s)
Phlebotomus/genetics , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Animals , Female , French Guiana , Gene Library , Male , Species SpecificityABSTRACT
Abstract INTRODUCTION: American cutaneous leishmaniasis (ACL) is regarded as a public health problem in the Oiapoque basin, between Brazil and French Guiana. METHODS Data on ACL occurrence/epidemiological profile and etiology were sourced from Brazilian health services and a reference laboratory. Rainfall correlation was also analyzed. RESULTS: Majority of the ACL cases were observed in adult men working as gold miners. ACL incidence peaks appeared to be linked to periods 2 months after the dry season. Migratory flow was found to be a non-negligible complicating factor in epidemiological surveillance. CONCLUSIONS: Binational strategies are required to minimize exposure for high-risk populations.
Subject(s)
Humans , Male , Female , Adult , Aged , Young Adult , Leishmaniasis, Cutaneous/epidemiology , Seasons , Brazil/epidemiology , Incidence , French Guiana/epidemiology , Middle AgedABSTRACT
BACKGROUND: An entomological study was conducted in the municipality of Oiapoque (lower Oyapock River Basin) in the Brazilian side bordering French Guiana to gain information on the transmission pattern of American cutaneous leishmaniasis (ACL) in that region, presumed to reflect the classical Amazonian/Guianan enzootic scenario. METHODS: Three ecologically isolated forested areas near urban environments were surveyed during the rainy and dry seasons of 2015 and 2016, using a multi-trapping approach comprising ground-level and canopy light traps, black and white colored cloth Shannon traps and manual aspiration on tree bases. Female phlebotomines were dissected to find infections and isolate flagellates from Leishmania spp. The strains were characterized by restriction fragment length polymorphism analysis and compared with those of local ACL cases and World Health Organization reference strains. RESULTS: Nyssomyia umbratilis, Trichopygomyia trichopyga and Evandromyia infraspinosa were the most frequently found species. Findings on relative abundance, spatiotemporal vector/ACL congruence, natural infections and anthropophilic insights strengthened the Guianan classical transmission of Leishmania (Viannia) guyanensis by Ny. umbratilis and suggested further investigations for Ev. infraspinosa. Nyssomyia umbratilis showed an eclectic feeding habit, including bird blood. Ecological data and literature reports also included Psychodopygus squamiventris maripaensis and Bichromomyia flaviscutellata on the list of suspected vectors. CONCLUSIONS: These findings contributed to understanding ACL ecoepidemiology in the Amazonian/Guianan scenario. Local studies are required to better comprehend the Leishmania spp. enzootic mosaic in specific ecotopes.
Subject(s)
Insect Vectors/parasitology , Leishmania/physiology , Leishmaniasis, Cutaneous/parasitology , Leishmaniasis, Cutaneous/transmission , Psychodidae/parasitology , Animals , Brazil , Ecosystem , Female , French Guiana , Humans , Insect Vectors/physiology , Leishmania/genetics , Leishmania/isolation & purification , Male , Psychodidae/physiology , SeasonsABSTRACT
BACKGROUND: New world cutaneous leishmaniasis (NWCL) can be found in French Guiana as well as in several other parts of Central and South America. Leishmania guyanensis accounts for nearly 90% of cases in French Guiana and is treated with pentamidine isethionate, given by either intramuscular or intravenous injection. The military population is particularly exposed due to repeated missions in the rainforest. The purpose of the present study was to identify the factors associated with pentamidine isethionate treatment failure in a series of service members with L. guyanensis NWCL acquired in French Guiana. METHOD: All the French service members reported as having acquired leishmaniasis in French Guiana from December 2013 to June 2016 were included. RESULTS: Seventy-three patients infected with L. guyanensis were included in the final analysis. Patients treated with IV pentamidine isethionate had better response rates than those treated with IM pentamidine isethionate (pâ¯=â¯0.002, adjusted odds ratio (AOR)â¯=â¯0.15, 95% CI [0.04-0.50]). The rate of treatment success was 85.3% (95% CI [68.9-95.0]) for IV pentamidine isethionate and 51.3% (95% CI [34.8-67.6]) for IM pentamidine isethionate. CONCLUSIONS: The use of intramuscular pentamidine isethionate in the treatment of Leishmania guyanensis cutaneous leishmaniasis is associated with more treatment failures than intravenous pentamidine isethionate.
Subject(s)
Antiprotozoal Agents/administration & dosage , Antiprotozoal Agents/adverse effects , Injections, Intramuscular/adverse effects , Leishmaniasis, Cutaneous/drug therapy , Pentamidine/administration & dosage , Pentamidine/adverse effects , Treatment Failure , Administration, Intravenous/adverse effects , Adult , Female , French Guiana/epidemiology , Humans , Leishmania guyanensis/drug effects , Leishmaniasis, Cutaneous/epidemiology , Male , Military Personnel , Odds Ratio , Pentamidine/therapeutic use , Risk Factors , South America/epidemiology , Treatment Outcome , Young AdultABSTRACT
INTRODUCTION: Leishmania RNA virus type 1 (LRV1) is an endosymbiont of some Leishmania (Vianna) species in South America. Presence of LRV1 in parasites exacerbates disease severity in animal models and humans, related to a disproportioned innate immune response, and is correlated with drug treatment failures in humans. Although the virus was identified decades ago, its genomic diversity has been overlooked until now. METHODOLOGY/PRINCIPLES FINDINGS: We subjected LRV1 strains from 19 L. (V.) guyanensis and one L. (V.) braziliensis isolates obtained from cutaneous leishmaniasis samples identified throughout French Guiana with next-generation sequencing and de novo sequence assembly. We generated and analyzed 24 unique LRV1 sequences over their full-length coding regions. Multiple alignment of these new sequences revealed variability (0.5%-23.5%) across the entire sequence except for highly conserved motifs within the 5' untranslated region. Phylogenetic analyses showed that viral genomes of L. (V.) guyanensis grouped into five distinct clusters. They further showed a species-dependent clustering between viral genomes of L. (V.) guyanensis and L. (V.) braziliensis, confirming a long-term co-evolutionary history. Noteworthy, we identified cases of multiple LRV1 infections in three of the 20 Leishmania isolates. CONCLUSIONS/SIGNIFICANCE: Here, we present the first-ever estimate of LRV1 genomic diversity that exists in Leishmania (V.) guyanensis parasites. Genetic characterization and phylogenetic analyses of these viruses has shed light on their evolutionary relationships. To our knowledge, this study is also the first to report cases of multiple LRV1 infections in some parasites. Finally, this work has made it possible to develop molecular tools for adequate identification and genotyping of LRV1 strains for diagnostic purposes. Given the suspected worsening role of LRV1 infection in the pathogenesis of human leishmaniasis, these data have a major impact from a clinical viewpoint and for the management of Leishmania-infected patients.
Subject(s)
Genetic Variation , Leishmania/virology , Leishmaniavirus/classification , Leishmaniavirus/isolation & purification , Phylogeny , Adult , Aged , Cluster Analysis , Female , French Guiana , Genome, Viral , Humans , Leishmania/isolation & purification , Leishmaniasis/parasitology , Leishmaniavirus/genetics , Male , Middle Aged , RNA, Viral/genetics , Sequence Alignment , Sequence Analysis, DNA , Young AdultABSTRACT
From 1996 to 1999 multi-trapping methods (Center of Diseases Control, CDC) light traps, light-baited Shannon traps, and aspiration on tree bases) were used to study the phlebotomine fauna of the "Serra do Navio" region of the Brazilian State of Amapá, which is part of the Guiana Shield. Fifty-three species were identified among 8,685 captured individuals. The following species, associated with the transmission of American cutaneous leishmaniasis in Amazonian Brazil, were captured: Nyssomyia umbratilis (3,388), Psychodopygus squamiventris maripaensis (995), Ny. anduzei (550), Trichophoromyia ubiquitalis (400), Ny. whitmani (291), Ps. paraensis (116), and Bichromomyia flaviscutellata (50). Flagellate infections were detected in 45 flies. Of the 19 parasites isolated in vitro, 15 were Leishmania (Viannia) guyanensis (13 in Ny. umbratilis, 1 in Ny. whitmani, 1 in Ny. anduzei) and three were L. (V.) naiffi (2 in Ps. s. maripaensis, 1 in Ny. anduzei). The results indicate the participation of three phlebotomine species in the transmission of L. (V.) guyanensis and two species in that of L. (V.) naiffi, and show that the same phlebotomine species is involved in the transmission of different Leishmania (Viannia) species in the Guianan/Amazon region. A review of the literature together with the results of the present study, and other published and unpublished results, indicate that eight phlebotomine species potentially participate in the transmission of Leishmania (Viannia) naiffi in Amazonia.
Subject(s)
Insect Vectors/parasitology , Leishmania/classification , Leishmaniasis, Cutaneous/transmission , Psychodidae/parasitology , Animals , Brazil , Climate , Female , Insect Vectors/classification , Isoenzymes/analysis , Leishmania/enzymology , Leishmania/genetics , Leishmania/isolation & purification , Male , Mining , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Psychodidae/classification , SeasonsABSTRACT
AbstractAnti-leishmaniasis drug resistance is a common problem worldwide. The aim of this study was to inventory the general in vitro level of sensitivity of Leishmania isolates circulating in French Guiana and to highlight potential in vitro pentamidine-resistant isolates. This sensitivity study was conducted on 36 patient-promastigote isolates for seven drugs (amphotericin B, azithromycin, fluconazole, meglumine antimoniate, miltefosine, paromomycin, and pentamidine) using the Cell Counting Kit-8 viability test. The IC50 values obtained were heterogeneous. One isolate exhibited high IC50 values for almost all drugs tested. Pentamidine, which is the first-line treatment in French Guiana, showed efficacy at very low doses (mean of 0.0038 µg/mL). The concordance of the in vitro pentamidine results with the patients' clinical outcomes was 94% (K = 0.82).
Subject(s)
Antiprotozoal Agents/pharmacology , Leishmania braziliensis/drug effects , Leishmania guyanensis/drug effects , Leishmaniasis, Cutaneous/drug therapy , Pentamidine/pharmacology , Amphotericin B/pharmacology , Azithromycin/pharmacology , Drug Resistance , Fluconazole/pharmacology , Humans , Leishmania braziliensis/growth & development , Leishmania braziliensis/isolation & purification , Leishmania guyanensis/growth & development , Leishmania guyanensis/isolation & purification , Leishmaniasis, Cutaneous/parasitology , Meglumine/pharmacology , Meglumine Antimoniate , Organometallic Compounds/pharmacology , Parasitic Sensitivity Tests , Paromomycin/pharmacology , Phosphorylcholine/analogs & derivatives , Phosphorylcholine/pharmacology , Treatment OutcomeABSTRACT
Treatment failure and symptomatic relapse are major concerns in American tegumentary leishmaniasis (TL). Such complications are seen frequently in Leishmania guyanensis infections, in which patients respond variously to first-line antileishmanials and are more prone to develop chronic cutaneous leishmaniasis. The factors underlying this pathology, however, are unknown. Recently, we reported that a double-stranded RNA virus, Leishmania RNA virus 1 (LRV1), nested within L. guyanensis parasites is able to exacerbate experimental murine leishmaniasis by inducing a hyperinflammatory response. This report investigates the prevalence of LRV1 in human L. guyanensis infection and its effect on treatment efficacy, as well as its correlation to symptomatic relapses after the completion of first-line treatment. In our cohort of 75 patients with a diagnosis of primary localized American TL, the prevalence of LRV1-positive L. guyanensis infection was elevated to 58%. All patients infected with LRV1-negative L. guyanensis were cured after 1 dose (22 of 31 [71%]) or 2 doses (31 of 31 [100%]) of pentamidine. In contrast, 12 of 44 LRV1-positive patients (27%) presented with persistent infection and symptomatic relapse that required extended therapy and the use of second-line drugs. Finally, LRV1 presence was associated with a significant increase in levels of intra-lesional inflammatory markers. In conclusion, LRV1 status in L. guyanensis infection is significantly predictive (P = .0009) of first-line treatment failure and symptomatic relapse and has the potential to guide therapeutic choices in American TL.