• Awareness
• HIV
• Immunosuppression
• Missed opportunity
• Visceral leishmaniasis

• Adult
• Female
• Humans
• Male
• Coinfection/drug therapy
• Delayed Diagnosis
• HIV Infections
• Immunosuppression Therapy
• Leishmaniasis, Visceral/diagnosis
• Leishmaniasis, Visceral/drug therapy
• Leishmaniasis, Visceral/epidemiology
• Retrospective Studies
• Middle Aged

• Leishmaniasis
• Molecular diagnosis
• Pan-Leishmania
• Recombinase polymerase amplification

• Humans
• DNA, Kinetoplast/genetics
• Leishmania/genetics
• Leishmania/isolation & purification
• Nucleic Acid Amplification Techniques/methods
• Recombinases/genetics
• RNA, Ribosomal, 18S/genetics
• Sensitivity and Specificity
• Leishmaniasis/diagnosis

Reports of leishmaniasis are scarce in North America. It is considered to be one of the neglected tropical diseases. It is seen in immigrants from endemic areas to United States. Treatments are not readily available in the United States. Untreated or inadequately treated cutaneous leishmaniasis not only causes localized disfigurement but can advance to more permanent and devastating mucosal disfigurement and perforation, if caused by a species that can also cause mucocutaneous leishmaniasis.

A 42-year-old human immunodeficiency virus negative male immigrant from Honduras presented to the emergency department of our facility in Louisiana with a 2-mo history of a left lower extremity ulcer. It started as a painless blister that progressed in size and developed into other smaller lesions tracking up the thigh and became tender and erythematous. Clinically looked nontoxic and healthy. He was afebrile. Blood tests, except inflammatory markers, were within normal limits. The cellulitis of the leg was treated with 6 d of vancomycin that also relieved the pain. Skin biopsy was obtained, and histopathology was suspicious for leishmania. Polymerase chain reaction/deoxyribonucleic acid sequencing done by centers for disease control and prevention confirmed the diagnosis as Leishmania panamensis. There was no involvement of naso-oropharyngeal mucosa, confirmed by otolaryngology. The patient was treated with miltefosine for 28 d. Clinic follow-up after approximately 11 mo revealed a healed skin ulcer.

Cutaneous leishmaniasis should be in the differential diagnosis of skin ulcers of travelers from endemic areas. Awareness regarding diagnosis and treatment of leishmaniasis needs to be enhanced.

• Cutaneous leishmaniasis
• Neglected diseases
• Leishmania (Viannia) panamensis
• Miltefosine
• Leishmania
• Case report

• Animals
• DNA/genetics
• Feeding Behavior
• Insect Vectors
• Leishmania/genetics
• Psychodidae/classification
• Psychodidae/genetics
• Psychodidae/pathogenicity
• Seasons
• Sicily
• Species Specificity
• Trypanosomatina/genetics

• Investigator Starting Grant
• PhD grant

In Colombia, the cutaneous leishmaniasis (CL) is the most common manifestation across the army personnel. Hence, it is mandatory to determine the species associated with the disease as well as the association with the clinical traits. A total of 273 samples of male patients with CL were included in the study and clinical data of the patients was studied. PCR and sequencing analyses (Cytb and HSP70 genes) were performed to identify the species and the intra-specific genetic variability. A georeferenced database was constructed to identify the spatial distribution of Leishmania species isolated. The identification of five species of Leishmania that circulate in the areas where army personnel are deployed is described. Predominant infecting Leishmania species corresponds to L. braziliensis (61.1%), followed by Leishmania panamensis (33.5%), with a high distribution of both species at geographical and municipal level. The species L. guyanensis, L. mexicana and L. lainsoni were also detected at lower frequency. We also showed the identification of different genotypes within L. braziliensis and L. panamensis. In conclusion, we identified the Leishmania species circulating in the areas where Colombian army personnel are deployed, as well as the high intra-specific genetic variability of L. braziliensis and L. panamensis and how these genotypes are distributed at the geographic level.

Colombia is one of the countries with the highest incidence of Cutaneous Leishmaniasis in the world and the army population is the most vulnerable population. Herein, we identified the infecting Leishmania species (L. braziliensis, L. panamensis, L. guyanensis, L. mexicana and L. lainsoni). We also showed the high intra-specific genetic variability of L. braziliensis and L. panamensis and how these genotypes are distributed at the geographic level.

• Adult
• Colombia/epidemiology
• DNA, Protozoan/chemistry
• DNA, Protozoan/genetics
• Genotype
• Humans
• Leishmania/classification
• Leishmania/isolation & purification
• Leishmaniasis, Cutaneous/epidemiology
• Leishmaniasis, Cutaneous/microbiology
• Male
• Military Personnel
• Phylogeography
• Polymerase Chain Reaction
• Sequence Analysis, DNA
• Topography, Medical
• Young Adult

• Departamento Administrativo de Ciencia, Tecnología e Innovación (CO)
• Dirección de Ciencia y Tecnología del Comando de Educación y Doctrina del ejército y al Comando de Personal del Ejército

• Animals
• Brazil
• Ecology
• Humans
• Insect Vectors
• Leishmania/isolation & purification
• Leishmaniasis/transmission
• Psychodidae/parasitology

• Fundação de Amparo a Pesquisa do Estado de Minas Gerais/FAPEMIG
• Conselho Nacional de Desenvolvimento Científico e Tecnológico/CNPq

• Culture
• Diagnostic
• Laboratory
• Molecular diagnosis
• NZD
• Neglected tropical disease
• Neglected zoonotic disease
• PCR
• Reliability
• Sensitivity
• Serology
• Specificity

Recently, there has been a re-emergence of cutaneous leishmaniasis in endemic countries and an increase in imported cases in non-endemic countries by travelers, workers, expatriates, immigrants, and military force personnel. Old World cutaneous leishmaniasis is caused primarily by Leishmania major, L. tropica and L. aethiopica. Despite their low sensitivity, diagnosis traditionally includes microscopic and histopathological examinations, and in vitro cultivation. Several conventional PCR techniques have been developed for species identification, which are time-consuming and labour-intensive. Real-time PCR using SYBR green dye, although provides rapid detection, may generate false positive signals. Therefore, a rapid and easy method such as a FRET-based real-time PCR would improve not only the turn-around time of diagnosing Old World cutaneous Leishmania species but will also increase its specificity and sensitivity.

A FRET-based real-time PCR assay which amplifies the cathepsin L-like cysteine protease B gene encoding a major Leishmania antigen was developed to differentiate L. major, L. tropica, and L. aethiopica in one single step using one set of primers and probes. Assay performance was tested on cutaneous and visceral strains of Leishmania parasite cultures and isolates of other protozoan parasites as well as human biopsy specimen.

The assay readily differentiates between the three Old World cutaneous leishmaniasis species based on their melting curve characteristics. A single Tm at 55.2 ± 0.5 °C for L. aethiopica strains was distinguished from a single Tm at 57.4 ± 0.2 °C for L. major strains. A double curve with melting peaks at 66.6 ± 0.1 °C and 48.1 ± 0.5 °C or 55.8 ± 0.6 °C was observed for all L. tropica strains. The assay was further tested on biopsy specimens, which showed 100 % agreement with results obtained from isoenzyme electrophoresis and Sanger sequencing.

Currently, there are no published data on real-time PCR using FRET technology to differentiate between Old World cutaneous Leishmania species. In summary, our assay based on specific hybridization addresses the limitations of previous PCR technology and provides a single step, reliable method of species identification and rapid diagnostic applications.

• Cutaneous
• Diagnosis
• FRET
• Leishmania
• Melting curve
• Real-time PCR
• qPCR

• Leishmania mexicana
• arginase
• α,α-disubstituted boronic amino-acid inhibitors

• Amino Acids/chemistry
• Animals
• Arginase/antagonists & inhibitors
• Arginase/chemistry
• Boronic Acids/chemistry
• Boronic Acids/pharmacology
• Enzyme Inhibitors/pharmacology
• Leishmania mexicana/enzymology
• Models, Molecular

• P30 EB009998 NIBIB NIH HHS
• R01 GM049758 NIGMS NIH HHS
• GM49758 NIGMS NIH HHS

• Citocromo b
• Cytochrome b
• Europe du sud-ouest
• Leishmania tropica
• PCR-RFLP
• Phlebotomus sergenti
• South-western Europe
• Sudoeste de Europa
• características ecológicas
• cytochrome b
• ecological traits
• lignées mitochondriales
• linajes mitocondriales
• mitochondrial lineages
• traits écologiques

• Adolescent
• Antimony Sodium Gluconate/therapeutic use
• Disease Outbreaks
• Female
• Humans
• Leishmania braziliensis/isolation & purification
• Leishmania guyanensis/isolation & purification
• Leishmaniasis, Cutaneous/drug therapy
• Leishmaniasis, Cutaneous/epidemiology
• Male
• Military Personnel
• Peru/epidemiology
• Real-Time Polymerase Chain Reaction
• Surveys and Questionnaires
• Young Adult

• D43 TW007393 FIC NIH HHS
• 2D43 TW007393 FIC NIH HHS

In the past decade, the number of imported leishmaniasis cases has increased in countries of Western Europe. The trend is associated with increasing travels, ecotourism activity, military operations and immigration. While in endemic countries leishmaniasis is usually well diagnosed, accurate patient history and parasite identification are necessary to distinguish between autochthonous and imported cases. This is particularly important, as new Leishmania species/genotypes may be introduced and transmitted by local phlebotomine vectors without appropriate surveillance, with unpredictable consequences. We report on the surveillance of imported leishmaniasis performed by the Leishmania Identification Reference Centre of Rome from 1986 through 2012, involving health care centres from 16/20 Italian regions. Suspected imported cases were analyzed and conclusions were based on clinical, epidemiological and diagnostic findings. Over the years, different parasite identification methods were employed, including MultiLocus Enzyme Electrophoresis and molecular techniques combining disease diagnosis (SSU rDNA nested-PCR) and Leishmania typing (nuclear repetitive sequence and ITS-1 PCR-RFLPs). A total of 105 imported cases were recorded (annual range: 0-20) of which 36 were visceral (VL) (16 HIV-coinfections) and 69 cutaneous (CL) cases; 85 cases (52 CL) were from the Old World and 20 (17 CL) from the New World. Eight Leishmania species were identified, of which 7 were exotic to Italy. VL importation until 1995 was associated with the spread of Mediterranean Leishmania-HIV co-infections in early 1990s. Following the introduction of HAART treatment, such cases became occasional in Italians but relatively frequent among immigrants. In contrast, a steady increase of CL cases was observed from different areas of the Old and New Worlds, that in recent years included mainly immigrants ‘visiting friends and relatives’ and Italian tourists. This positive trend likely depends on better diagnosis and reporting; however, we suspect that many CL cases remained unrecognized. Given the relatively low incidence of leishmaniasis importation, the risk of introduction of exotic parasites appears limited, although the detection of anthroponotic species requires attention.

• Humans
• Incidence
• Italy/epidemiology
• Leishmania/classification
• Leishmania/genetics
• Leishmaniasis/epidemiology
• Leishmaniasis/parasitology
• Leishmaniasis/transmission
• Population Surveillance
• Travel

Cutaneous leishmaniasis caused by Leishmania major is an emergent, uncontrolled public health problem and there is no vaccine. A promising prophylactic approach has been immunotherapy with Toll-like receptor (TLR) agonists to enhance parasite-specific immune responses. We have previously reported that vaccination of C57BL/6 mice with live L. major plus the TLR9 agonist CpG DNA prevents lesion development and confers immunity to reinfection. Our current study aims to investigate whether other TLR agonists can be used in leishmanization without induction of lesion formation. We found that live L. major plus the TLR2 agonist Pam3CSK4 reduced the pathology in both genetically resistant (C57BL/6) and susceptible (BALB/c) mouse strains. The addition of Pam3CSK4 activated dermal dendritic cells and macrophages to produce greater amounts of proinflammatory cytokines in both mouse strains. Both Th1 and Th17 responses were enhanced by leishmanization with L. major plus Pam3CSK4 in C57BL/6 mice; however, Th17 cells were unchanged in BALB/c mice. The production of IL-17 from neutrophils was enhanced in both strains infected with L. major plus Pam3CSK4. However, the sustained influx of neutrophils in sites of infection was only observed in BALB/c mice. Our data demonstrate that the mechanism behind leishmanization with TLR agonists may be very different depending upon the immunological background of the host. This needs to be taken into account for the rational development of successful vaccines against the disease.

Cutaneous leishmaniasis is a skin infection caused by a protozoan parasite Leishmania major (L. major). The only available treatment option is chemotherapy, which is toxic and expensive. Currently, there is no vaccine. Although inoculation of virulent L. major (leishmanization) that provides effective protection in humans was widely applied, it was discontinued due to safety concerns. To improve the safety of leishmanization, we applied agonists of Toll-like receptor in the leishmanization to induce parasite-specific immune responses. In particular, we show here that inoculation with live L. major plus a TLR2 agonist Pam3CSK4 in both resistant (C57BL/6) and susceptible (BALB/c) mouse strains completely prevents the development of lesion and decreases parasite burden. The improved pathology is associated with enhanced production of IL-6 and IL-12 from dermal dendritic cells and macrophages. Both Th1 and Th17 responses are enhanced in C57BL/6 mice. Although only the Th1 response was enhanced in BALB/c mice in the presence of Pam3CSK4, there is an enhanced and sustained neutrophil influx at sites of infection. Overall, our study reveals the clinical significance of TLR2 agonist in treating cutaneous leishmaniasis. However, the protective mechanism may be quite different depending upon the genetic background of the host.

• Animals
• Cytokines/biosynthesis
• Dendritic Cells/parasitology
• Injections
• Leishmaniasis, Cutaneous/drug therapy
• Leishmaniasis, Cutaneous/immunology
• Leishmaniasis, Cutaneous/pathology
• Lipopeptides/administration & dosage
• Lipopeptides/therapeutic use
• Mice
• Mice, Inbred BALB C
• Mice, Inbred C57BL
• Toll-Like Receptor 2/agonists

• R21 AI061379 NIAID NIH HHS
• R21-AI-61379 NIAID NIH HHS

Leishmaniasis is a vector borne disease caused by protozoa of the genus Leishmania. Human leishmaniasis is not endemic in Australia though imported cases are regularly encountered. This study aimed to provide an update on the molecular epidemiology of imported leishmaniasis in Australia. Of a total of 206 biopsies and bone marrow specimens submitted to St Vincent’s Hospital Sydney for leishmaniasis diagnosis by PCR, 55 were found to be positive for Leishmania DNA. All PCR products were subjected to restriction fragment length polymorphism analysis for identification of the causative species. Five Leishmania species/species complexes were identified with Leishmania tropica being the most common (30/55). Travel or prior residence in a Leishmania endemic region was the most common route of acquisition with ~47% of patients having lived in or travelled to Afghanistan. Cutaneous leishmaniasis was the most common manifestation (94%) with only 3 cases of visceral leishmaniasis and no cases of mucocutaneous leishmaniasis encountered. This report indicates that imported leishmaniasis is becoming increasingly common in Australia due to an increase in global travel and immigration. As such, Australian clinicians must be made aware of this trend and consider leishmaniasis in patients with suspicious symptoms and a history of travel in endemic areas. This study also discusses the recent identification of a unique Leishmania species found in native kangaroos and a potential vector host which could create the opportunity for the establishment of a local transmission cycle within humans.

Leishmaniasis is increasingly reported among travellers. Leishmania species vary in sensitivity to available therapies. Fast and reliable molecular techniques have made species-directed treatment feasible. Many treatment trials have been designed poorly, thus developing evidence-based guidelines for species-directed treatment is difficult. Published guidelines on leishmaniasis in travellers do not aim to be comprehensive or do not quantify overall treatment success for available therapies. We aimed at providing comprehensive species-directed treatment guidelines.

English literature was searched using PubMed. Trials and observational studies were included if all cases were parasitologically confirmed, the Leishmania species was known, clear clinical end-points and time points for evaluation of treatment success were defined, duration of follow-up was adequate and loss to follow-up was acceptable. The proportion of successful treatment responses was pooled using mixed effects methods to estimate the efficacy of specific therapies. Final ranking of treatment options was done by an expert panel based on pooled efficacy estimates and practical considerations. 168 studies were included, with 287 treatment arms. Based on Leishmania species, symptoms and geography, 25 clinical categories were defined and therapy options ranked. In 12/25 categories, proposed treatment agreed with highest efficacy data from literature. For 5/25 categories no literature was found, and in 8/25 categories treatment advise differed from literature evidence. For uncomplicated cutaneous leishmaniasis, combination of intralesional antimony with cryotherapy is advised, except for L. guyanensis and L. braziliensis infections, for which systemic treatment is preferred. Treatment of complicated (muco)cutaneous leishmaniasis differs per species. For visceral leishmaniasis, liposomal amphotericin B is treatment of choice.

Our study highlights current knowledge about species-directed therapy of leishmaniasis in returning travellers and also demonstrates lack of evidence for treatment of several clinical categories. New data can easily be incorporated in the presented overview. Updates will be of use for clinical decision making and for defining further research.

Human leishmaniasis is caused by unicellular parasites that are injected into the skin by sand-flies, small, flying insects. Many different Leishmania species cause various manifestations of disease, both of the skin and internal organs. Leishmaniasis is a curable disease but clear guidelines on the best available treatment are lacking. Leishmania species differ in sensitivity to available drugs. Until recently, identification of the infecting Leishmania parasite was laborious, thus therapy could not precisely be targeted to the infecting species, in contrast to many other infectious diseases. Nowadays, Leishmania parasites can be identified relatively easily with new DNA techniques. We studied efficacy of therapies for diseases due to different Leishmania species, limited to the English literature. Efficacy was summarized and presented in an easy to read format. Because of difficulties with identification of parasite species in earlier studies, quality of evidence was often limited. Our findings are a major help for clinicians to easily find optimal treatment for specific patients. Moreover, our results demonstrate where additional research is needed to further improve treatment of leishmaniasis.

• global change
• human activities
• infectious disease
• natural factors

Leishmaniasis is a virulent parasitic infection that causes a worldwide disease burden. Most treatments have toxic side-effects and efficacy has decreased due to the emergence of resistant strains. The outlook is worsened by the absence of promising drug targets for this disease. We have taken a computational approach to the detection of new drug targets, which may become an effective strategy for the discovery of new drugs for this tropical disease.

We have predicted the protein interaction network of Leishmania major by using three validated methods: PSIMAP, PEIMAP, and iPfam. Combining the results from these methods, we calculated a high confidence network (confidence score > 0.70) with 1,366 nodes and 33,861 interactions. We were able to predict the biological process for 263 interacting proteins by doing enrichment analysis of the clusters detected. Analyzing the topology of the network with metrics such as connectivity and betweenness centrality, we detected 142 potential drug targets after homology filtering with the human proteome. Further experiments can be done to validate these targets.

We have constructed the first protein interaction network of the Leishmania major parasite by using a computational approach. The topological analysis of the protein network enabled us to identify a set of candidate proteins that may be both (1) essential for parasite survival and (2) without human orthologs. These potential targets are promising for further experimental validation. This strategy, if validated, may augment established drug discovery methodologies, for this and possibly other tropical diseases, with a relatively low additional investment of time and resources.

• Adjuvants, Immunologic/therapeutic use
• Animals
• Cells, Cultured
• CpG Islands/immunology
• Leishmania major/immunology
• Leishmaniasis, Cutaneous/immunology
• Leishmaniasis, Cutaneous/therapy
• Mice
• Mice, Inbred C57BL
• Mice, Knockout
• Oligodeoxyribonucleotides/immunology
• Oligodeoxyribonucleotides/therapeutic use
• Protozoan Vaccines/immunology
• Protozoan Vaccines/therapeutic use
• Vaccines, Attenuated/immunology
• Vaccines, Attenuated/therapeutic use
• Vaccines, DNA/immunology
• Vaccines, DNA/therapeutic use

• R01AI057992-01 NIAID NIH HHS
• R21AI61379 NIAID NIH HHS

• Animals
• Communicable Disease Control
• Disease Reservoirs
• Dog Diseases/parasitology
• Dogs
• Europe
• Host-Parasite Interactions
• Humans
• Insecticides
• Leishmaniasis/transmission
• Protozoan Vaccines/administration & dosage
• Psychodidae
• United States
• Zoonoses