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Background: We previously identified a Leishmania (L.) major large RAB GTPase (LmlRAB), a new atypical RAB GTPase protein. It is highly conserved in Leishmania species while displaying low level of homology with mammalian homologues. Leishmania small RAB GTPases proteins have been involved in regulation of exocytic and endocytic pathways whereas the role of large RAB GTPases proteins has not been characterized yet. We report here the immunogenicity of both recombinant rLmlRAB and rLmlRABC, in individuals with immunity against L. major or L. infantum.Methods: PBMC were isolated from individuals cured of L. major (CCLm) or from healthy individuals. The latter were subdivided into high or low IFN-γ responders. Healthy high IFN-γ responders, considered as asymptomatics, were living in an endemic area for L. major (HHRLm) or L. infantum (HHRLi). Healthy low IFN-γ responders (HLR) were considered as naïve controls. Cells from all volunteers were stimulated with rLmlRAB or rLmlRABC. Cytokines were analysed by CBA and ELISA and phenotypes of IFN-γ-producing cells were analysed by flow cytometry.Results: Both rLmlRAB and rLmlRABC induced high significant levels of IFN-γ in CCLm, HHRLm and HHRLi groups. Phenotype analysis of rLmlRAB and rLmlRABC-stimulated T cells in CCLm individuals showed a significant increase in the percentage of specific IFN-γ-producing CD4+ and CD8+ T cells. rLmlRAB induced significant granzyme B levels in CCLm and HHRLm. Low but significant granzyme B levels were detected in naïve group. IL-10 was detected in immune and naïve individuals.Conclusion: We showed that rLmlRAB protein and its divergent carboxy-terminal part induced a predominant Th1 response in individuals immune to L. major or L. infantum. Our results suggest that rLmlRAB and rLmlRABC proteins are potential cross-species vaccine candidates against cutaneous and visceral leishmaniasis.

Skin leishmaniasis is a transmissible disease, endemic in countries with warm and hot climates. The increase in the incidence and the frequency of complications, the lack of modern diagnostics determine the relevance of the problem of therapy of the disease. The use of polymerase chain reaction (PCR) is the best method of diagnosing leishmaniasis, since conventional methods (parasitological) are not sensitive enough.Objective — to evaluate the use of polymerase chain reaction for the diagnosis of type identification of pathogens of skin leishmaniasis.Materials and methods. The initial material for genetic analysis were swabs from epithelial tissues of the skin of the patient with skin leishmaniasis. The first step was identification of DNA of leishmania with determination of the DNA concentra­tion and clearness of DNA on the flowmeter. Next, allelespecific PCR was conducted to determine the type specificity of leishmanias. Examination of 40 samples of the biological material with skin leishmaniasis was performed. PCR was performed in regimen of real time (40 performances per every type of leishmania).Results and discussion. The performance of PCR in the regimen of real time provided identification of L. infantum in all 40 samples, while L. major and L. tropica were not registered. The isolation of L. infantum showed some epidemiological features connected to these clinical forms of the skin leishmaniosis, as well as allowed performing further type specific treatment, which will promote considerable increase of the general therapeutic effect in this disease.Conclusions. The investigations performed will allow the development of a more precise test, that is, a native panel of diagnosticums for identification of leishmania and its various types. The developed method of PCR-diagnosis in the real time may be used in the screening investigations of the population of the endemic zones of the republic., Кожный лейшманиоз — трансмиссивная болезнь, эндемичная в странах с теплым и жарким климатом. Рост заболеваемости, увеличение частоты осложнений, отсутствие современной диагностики обус­ловливают актуальность проблемы терапии данного заболевания. Использование полимеразной цепной реакции (ПЦР) — наиболее предпочтительный метод диагностики лейшманиоза, так как обычные методы (паразитологические) не являются достаточно чувствительными.Цель работы — оценить использование ПЦР для диагностики видовой идентификации возбудителей кожного лейшманиоза.Материалы и методы. Исходным материалом для генетического анализа служили соскобы из эпителиальной ткани кожи больного кожным лейшманиозом. Вначале выделяли ДНК лейшманий с определением концентрации и чистоты ДНК на флуорометре. Далее проводилась постановка аллельспецифической ПЦР для определения видовой специфичности лейшманий. Было проведено обследование 40 образцов биологического материала у больных кожным лейшманиозом, причем ПЦР проводилась в режиме реального времени (по 40 постановок на каждый вид лейшманий).Результаты и обсуждение. Проведение ПЦР в режиме реального времени позволило во всех 40 образцах выделить L. infantum, в то время как L. major и L. tropica не регистрировались. Выделение L. infantum указывает на некоторые эпидемиологические особенности, связанные с данными клиническими формами кожного лейшманиоза, а также позволяет в дальнейшем проводить видоспецифическое лечение, что позволит значительно повысить общий терапевтический эффект при данном заболевании.Выводы. Проведенные исследования позволят создать более точный тест — отечественную панель диагностикумов для обнаружения лейшманий и ее разновидностей. Разработанный метод ПЦР-диагностики в реальном времени может быть использован в скрининговых обследованиях населения эндемических зон республики., Шкірний лейшманіоз — трансмісивна хвороба, ендемічна в країнах з теплим і жарким кліматом. Зростання захворюваності, збільшення частоти ускладнень, відсутність сучасної діагностики зумовлюють актуальність проблеми терапії цього захворювання. Використання полімеразної ланцюгової реакції (ПЛР) — найкращий метод діагностики лейшманіозу, оскільки звичайні методи (паразитологічні) не є достатньо чутливими.Мета роботи — оцінити використання ПЛР для діагностики видової ідентифікації збудників шкірного лейшманіозу.Матеріали та методи. Вихідним матеріалом для генетичного аналізу слугували зіскрібки з епітеліальної тканини шкіри хворого на шкірний лейшманіоз. Спочатку виділяли ДНК лейшманій з визначенням концентрації і чистоти ДНК на флуорометрі. Далі проводилася постановка алельспецифічної ПЛР для визначення видової специфічності лейшманій. Було проведено обстеження 40 зразків біологічного матеріалу у хворих на шкірний лейшманіоз, причому ПЛР проводилася в режимі реального часу (по 40 постановок на кожен вид лейшманій).Результати та обговорення. Проведення ПЛР в режимі реального часу дало змогу в усіх 40 зразках виділити L. infantum, у той час як L. major і L. tropica не реєструвалися. Виділення L. infantum вказує на деякі епідеміологічні особливості, пов’язані з цими клінічними формами шкірного лейшманіозу, а також дозволяє в подальшому проводити видоспецифічне лікування, що дасть можливість значно підвищити загальний терапевтичний ефект при цьому захворюванні.Висновки. Проведені дослідження дають змогу створити більш точний тест — вітчизняну панель діагностикумів для виявлення лейшманій і її різновидів. Розроблений метод ПЛР­-діагностики в реальному часі може бути використаний в скринінгових обстеженнях населення ендемічних зон республіки.

We report here a case of simultaneous cutaneous and visceral manifestations due toLeishmania L. infantumdiagnosed in an immunocompetent adult. We describe a 74-year-old woman from Tunis, Tunisia, who presented a biologically confirmed visceral leishmaniasis infection concomitant with arm ulceration which appeared 2 years before.LeishmaniaDNA was detected by ITS PCR in both buffy coat and dermal scrapping of the arm lesion. Sequencing revealed that the 2 isolated strains corresponded toL. infantumand were 100% identical. The symptoms of visceral leishmaniasis responded to amphotericin B with rapid healing. However, the skin lesion did not improve althoughLeishmaniaPCR on dermal sample became negative. This location is probably secondarily to lymphatic or blood dissemination during the systemic visceral leishmaniasis infection. It would be favored by the inflammatory environment induced by the basal cell carcinoma subsequently diagnosed.

This study's main objective was to evaluate the action of larval therapy derived from Lucilia sericata and Sarconesiopsis magellanica (blowflies) regarding Leishmania panamensis using an in vivo model. Eighteen golden hamsters (Mesocricetus auratus) were used; they were divided into 6 groups. The first three groups consisted of 4 animals each; these, in turn, were internally distributed into subgroups consisting of 2 hamsters to be used separately in treatments derived from each blowfly species. Group 1 was used in treating leishmanial lesions with larval therapy (LT), whilst the other two groups were used for evaluating the used of larval excretions and secretions (ES) after the ulcers had formed (group 2) and before they appeared (group 3). The three remaining groups (4, 5 and 6), consisting of two animals, were used as controls in the experiments. Biopsies were taken for histopathological and molecular analysis before, during and after the treatments; biopsies and smears were taken for assessing parasite presence and bacterial co-infection. LT and larval ES proved effective in treating the ulcers caused by the parasite. There were no statistically significant differences between the blowfly species regarding the ulcer cicatrisation parameters. There were granulomas in samples taken from lesions at the end of the treatments. The antibacterial action of larval treatment regarding co-infection in lesions caused by the parasite was also verified. These results potentially validate effective LT treatment against cutaneous leishmaniasis aimed at using it with humans in the future. © 2016 Elsevier B.V.

Antonia MR Franco,1 Iryna Grafova,2 Fabiane V Soares,1,3 Gennaro Gentile,4 Claudia DC Wyrepkowski,1,3 Marcos A Bolson,5 Ézio Sargentini Jr,5 Cosimo Carfagna,4 Markku Leskelä,2 Andriy Grafov2 1Laboratory of Leishmaniasis and Chagas Disease, National Institute of Amazonian Research (INPA), Manaus, Amazonas, Brazil; 2Department of Chemistry, University of Helsinki, Helsinki, Finland; 3Multi-Institutional Post-Graduate Program in Biotechnology, Federal University of Amazonas, Manaus, Amazonas, Brazil; 4Institute for Polymers, Composites, and Biomaterials, National Research Council, Pozzuoli, Naples Province, Italy; 5Laboratory of Environmental Chemistry, National Institute of Amazonian Research (INPA), Manaus, Amazonas, Brazil Background: Coordination compounds of pentavalent antimony have been, and remain, the first-line drugs in leishmaniasis treatment for >70years. Molecular forms of Sb (V) complexes are commercialized as sodium stibogluconate (Pentostam®) and meglumine antimoniate (MA) (Glucantime®). Ever-increasing drug resistance in the parasites limits the use of antimonials, due to the low drug concentrations being administered against high parasitic counts. Sb5+ toxicity provokes severe side effects during treatment. To enhance therapeutic potency and to increase Sb (V) concentration within the target cells, we decided to try a new active substance form, a hydrosol of Sb2O5⋅nH2Onanoparticles (NPs), instead of molecular drugs. Methodology/principal findings: Sb2O5⋅nH2O NPs were synthesized by controlled SbCl5 hydrolysis in a great excess of water. Sb2O5⋅nH2Ophase formation was confirmed by X-ray diffraction. The surface of Sb (V) NPs was treated with ligands with a high affinity for target cell membrane receptors. The mean particle size determined by dynamic light scattering and transmission electron microscopy was ~35–45nm. In vitro tests demonstrated a 2.5–3 times higher antiparasitic activity of Sb (V) nanohybrid hydrosols, when compared to MA solution. A similar comparison for in vivo treatment of experimental cutaneous leishmaniasis with Sb5+ nanohybrids showed a 1.75–1.85 times more effective decrease in the lesions. Microimages of tissue fragments confirmed the presence of NPs inside the cytoplasm of infected macrophages. Conclusion/significance:Sb2O5⋅nH2Ohydrosols are proposed as a new form of treatment for cutaneous leishmaniasis caused by Leishmania amazonensis. The NPs penetrate directly into the affected cells, creating a high local concentration of the drug, a precondition to overcoming the parasite resistance to molecular forms of pentavalent antimonials. The nanohybrids are more effective at a lower dose, when compared to MA, the molecular drug. Our data suggest that the new form of treatment has the potential to reduce and simplify the course of cutaneous leishmaniasis treatment. At the same time, Sb2O5⋅nH2Ohydrosols provide an opportunity to avoid toxic antimony (V) spreading throughout the body. Keywords: nanoparticle, leishmaniasis, hydrated antimony (V) oxide, TEM, transmission electron microscopy

In this study, we identified the phlebotomine sandfly vectors involved in the transmission of American Cutaneous Leishmaniasis (ACL) in Assis Brasil, Acre, Brazil, which is located on the Brazil-Peru-Bolivia frontier. The genotyping of Leishmania in phlebotomines was performed using polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism. A total of 6,850 sandflies comprising 67 species were captured by using CDC light traps in rural areas of the municipality. Three sandfly species were found in the state of Acre for the first time: Lutzomyia georgii, Lu. complexa and Lu. evangelistai. The predominant species was Lu. auraensis/Lu. ruifreitasi and Lu. davisi (total 59.27%). 32 of 368 pools were positive for the presence of Leishmania DNA (16 pools corresponding to Lu. davisi, and 16 corresponding to Lu. auraensis/Lu. ruifreitasi), with a minimal infection prevalence of 1.85% in Lu. davisi and 2.05% in Lu. auraensis/Lu. ruifreitasi. The Leishmania species found showed maximum identity with L. (Viannia) guyanensis and L. (V.) braziliensis in both phlebotomine species. Based on these results and similar scenarios previously described along the Brazil/Peru/Bolivia tri-border, the studied area must take into consideration the possibility of Lu. davisi and Lu. auraensis/Lu. ruifreitasi as probable vectors of ACL in this municipality. © 2016, Fundacao Oswaldo Cruz. All rights reserved.

Background: The parasite of the genus Leishmania causes a neglected disease known as leishmaniasis, which has different clinical aspects depending on the species that infects the person and the immune response of the patient. The objective was to determine, using molecular biology, the current circulating species of Leishmania in biopsies of different types of cutaneous leishmaniasis (CL) from the endemic district in the region of “Madre de Dios”, Peru. Methods: The study's 19 samples were obtained in the Huepethue district in the “Madre de Dios” region from patients who were diagnosed for suspected leishmaniasis infection with three clinical manifestations. These were analyzed using kDNA-PCR, Nested-PCR and HRM-PCR to identify the genus of an infecting parasite as well as its species. Results: kDNA-PCR detected all tests for the genus of Leishmania to be positive, while the Nested-PCR could not detect 20.9% of species (one L. (V.) braziliensis and three L. (V.) amazonensis), and the HRM-PCR detected all species presented in the samples. The most prevalent species was L. (V.) braziliensis (73.7%), and the clinical manifestations were ulcers (63.2%), superficial scabs (5.3%) and diffuse form (5.3%), followed by L. (V.) amazonensis (15.8%), manifesting ulcers (15.6%), and L. (V.) lainsoni (10.5%), manifesting superficial scabs (5.3%) and ulcers (5.3%). L. (V.) braziliensis, L. (V.) amazonensis and L. (V.) lainsoni were detected effectively using HRM-PCR in the samples with different clinical manifestations. Conclusions: Highlighting the findings of a high diversity of Leishmania species using High Resolution Melting PCR in biopsies with different clinical manifestations of cutaneous leishmaniasis. Keywords: Leishmania, Clinical manifestations, PCR, Genotyping, Cutaneous leishmaniasis

Aim: Here, we evaluate the ddPCR platform using an evaluated qPCR-based diagnostic assay for the detection of Leishmania infection in Cutaneous Leishmaniasis patients. Methods: A standard curve of cultured Leishmania parasite material and clinical samples of CL patients were tested with ddPCR to determine the analytical and diagnostic performance. Results: The limit of detection of the assay on the ddPCR platform was much higher than the published limit of detection of the same assay on the qPCR platform (100 vs 1 parasites/mL, respectively). Conclusion: While the performance of this assay in ddPCR format was acceptable for research purposes, it is not sufficient for clinical diagnostic purposes. The assay is more suited to the qPCR platform. Keywords: Leishmania, ddPCR, Molecular diagnosis, Cutaneous Leishmaniasis

Leishmaniasis is a disease caused by intracellular protozoan parasites of the genus Leishmania. In endemic areas, only a portion of exposed subjects develops cutaneous leishmaniasis (CL), suggesting that the genetic inheritance of the host plays a vital role in both resistance and susceptibility to the disease. Interleukin-2 (IL-2) is a cytokine that plays a central role in the regulation of the immune response in infection through the axis IL-2/IL-2R (receptor) complex, triggering a series of intracellular events, among which the signaling of Janus kinase/signal transducers and activators of transcription (JAK-STAT). The present study aimed at verifying the possible relationship between single nucleotide polymorphism (s) (SNP s) in the genes IL-2, IL-2RB, and JAK3 in subjects with CL caused by Leishmania guyanensis in the city of Manaus, state of Amazonas, Brazil. 820 patients with CL and 850 healthy subjects (control group) coming from the same endemic areas as the patients were examined. The SNPs -2425G/A (rs4833248) and -330 T/G (rs2069762), located in the IL-2 gene promoter region, seem to influence the expression of the gene and the SNP +10558G/A (rs1003694) and +13295T/C (rs3212760) located in the 3rd intron of the IL-2RB gene and the 13th intron of the JAK3 gene, respectively, were studied by PCR-RFLP. Genotypes and alleles frequencies were obtained by direct counting. For the comparison between the two groups, the χ2 test with OR (odds ratio) and the 95% confidence interval (CI) were used. Similar genotypes and alleles frequencies for the different SNPs were observed in both patients with CL and healthy controls. Comparison of genotypic and allelic frequency between patients with CL and healthy subjects did not show any difference. These polymorphisms do not predict susceptibility to, or protection against the development of CL caused by L. guyanensis in the Amazonas. © 2019 de Araújo Santos et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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., Author summary Cutaneous leishmaniasis is a vector-borne zoonotic disease with a complex transmission cycle that includes many parasite, vector and host species. This disease continues to pose public health problems worldwide despite the measures put in place. In recent years, methodological tools commonly used in ecology, called ecological niche prediction models, have made it possible to determine the environmental and anthropogenic variables that may be favourable to the presence of the host and vector species communities involved in the cycle and therefore to the presence of certain disease agents. The use of these models, based on the presence of human cases of the disease, can overcome some of the uncertainties concerning the diversity of the vectors and the potential hosts involved in the transmission cycle. This approach of health ecology combining ecology and epidemiology could provide new insights into understanding the cycle of disease transmission and the influence of environmental factors and thus improve the prediction of disease emergence and epidemics. It can be applied to various vector-borne diseases whose transmission cycles are still poorly understood and for which studies classically carried out in epidemiology have not prevented disease progression.

Loop-mediated isothermal amplification (LAMP) is ideal for the detection of Leishmania DNA as it is a quick and easy-to-perform test that does not require complex or sophisticated equipment or infrastructure. However, the application of this technique in the detection of Leishmania DNA has not been comprehensively analyzed to date (analytical validation). Our objective was to evaluate the sensitivity and analytical specificity (anticipated reportable range [ARR], the limit of detection [LoD], and accuracy) of LAMP targeting the 18S rRNA gene in the diagnosis of six New World Leishmania species. We then applied the validated LAMP assay across 50 samples of sandflies and 50 direct smears from a recent outbreak of cutaneous leishmaniasis in Colombia to determine its diagnostic performance. The LAMP assay exclusively amplified the DNA of Leishmania spp., and an ARR of between 1 × 10 4 and 1 × 10 -2 equivalent parasites/mL was determined. An LoD of 1 × 10 -2 equivalent parasites/mL was established and there was no statistically significant variation in terms of accuracy. Finally, a sensitivity of 100% in direct smears and sandflies samples was calculated and a specificity of 90.9% for direct smears using microscopy as reference and 96.8% for sandflies using real-time polymerase chain reaction as reference were determined. To our knowledge, this is the first attempt to analytically validate a LAMP test to detect Leishmania DNA, which showed good diagnostic potential from sandflies and direct smear samples. Copyright © 2018 by The American Society of Tropical Medicine and Hygiene.

To evaluate the dynamics of regulatory T cells (Tregs) during tegumentary leishmaniasis, we assessed peripheral blood and biopsies from 54 patients. Patients with cutaneous leishmaniasis (CL) had a decreased proportion of Tregs in the peripheral blood, but the proportion was higher in the biopsies of lesions. During treatment of CL, circulating Tregs increased reaching normal proportions, whereas antigen-specific interferon-γ responses diminished. By contrast, circulating Tregs from mucosal leishmaniasis patients failed to normalize during treatment. C-C chemokine receptor type 5 was expressed on a large proportion of Tregs at the site of infection. These results demonstrate increased Tregs at the site of infection, possibly homing from the peripheral circulation.

Background: The serological diagnostic methods currently available for mucocutaneous leishmaniasis (MCL) lack specificity when complete parasites are used; however, such specificity increases when protein fractions are used. Ribosomal proteins have been reported to induce antibodies in animal and humans infected with the parasite, making them a worth candidate to assess its diagnosis potential. Objective: This study was thus aimed at evaluating synthetic peptides derived from Leishmania braziliensis ribosomal proteins S25 and S5 as antigen candidates for diagnosing MCL by ELISA Methods: It was used 8 and 13 peptides derived from ribosomal proteins 25 and S5 respectively as antigens in order to detect IgG antibodies by ELISA in people with active MCL, Chagas disease (CH) and autoimmune disease (AID). Results: 4 of these 21 peptides (P4, P6, P19 and P21) had the greatest sensitivity (21.7%, 13.04%, 20% and 20%, respectively) as well as having 95%, 100%, 100% and 82.5% specificity, respectively. Conclusion: The study revealed the limited usefulness of the peptides being studied as a diagnostic tool in the conditions used here, because its low sensitivity, but it is worth highlighting that the use of peptides as antigen in the serodiagnosis of MCL may overcome the cross reaction presented with other antigens, thus avoiding false positives.

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., Author summary 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.

The data on long-term efficacy, safety and drug survival rates of conventional systemic therapeutics in pediatric psoriasis is lacking. The primary aim of this study is to investigate acitretin, methotrexate, cyclosporin efficacy, safety and drug survival rates in pediatric patients as well as predictors of drug survival. This is a multicenter study including 289 pediatric cases being treated with acitretin, methotrexate and cyclosporin in four academic referral centers. Efficacy, adverse events, reasons for discontinuation, 1, 2- and 3-year drug survival rates, and determinants of drug survival were analyzed. A 75% reduction of Psoriasis Area and Severity Index score or better response rate was obtained in 47.5%, 34.1% and 40% of the patients who were treated with acitretin, methotrexate and cyclosporin, respectively. One-year drug survival rates for acitretin, methotrexate and cyclosporin were 36.3%, 21.1% and 15.1%, respectively. The most significant determinant of drug survival, which diminished over time, was treatment response whereas arthritis, body mass index and sex had no influence. Although all three medications are effective and relatively safe in children, drug survival rates are low due to safety concerns at this age group. Effective disease control through their rational use can be expected to improve survival rates. Pfizer Novartis Merck & Company - Schering Plough Corporation AbbVie Merck & Company Johnson & Johnson - Johnson & Johnson USA - Janssen Biotech Inc

The kinetoplastids include a large number of parasites responsible for serious diseases in humans and animals (Leishmania and Trypanosoma brucei) considered endemic in several regions of the world. These parasites are characterized by digenetic life cycles that undergo morphological and genetic changes that allow them to adapt to different microenvironments on their vertebrates and invertebrates hosts. Recent advances in ´omics´ technology, specifically transcriptomics have allowed to reveal aspects associated with such molecular changes. So far, different techniques have been used to evaluate the gene expression profile during the various stages of the life cycle of these parasites and during the host-parasite interactions. However, some of them have serious drawbacks that limit the precise study and full understanding of their transcriptomes. Therefore, recently has been implemented the latest technology (RNA-seq), which overcomes the drawbacks of traditional methods. In this review, studies that so far have used RNA-seq are presented and allowed to expand our knowledge regarding the biology of these parasites and their interactions with their hosts. © 2017 Elsevier B.V.

In order to understand the epidemiological dynamics of antimonial (SbV) resistance in zoonotic tegumentary leishmaniasis and its link with treatment outcome, we analyzed the population structure of 24 Peruvian Leishmania braziliensis clinical isolates with known in vitro antimony susceptibility and clinical phenotype by multilocus microsatellite typing (14 microsatellite loci). The genetic variability in the Peruvian isolates was high and the multilocus genotypes were strongly differentiated from each other. No correlation was found between the genotypes and in vitro drug susceptibility or clinical treatment outcome. The finding of a polyphyletic pattern among the SbV-resistant L. braziliensis might be explained by (i) independent events of drug resistance emergence, (ii) sexual recombination and/or (iii) other phenomena mimicking recombination signals. Interestingly, the polyphyletic pattern observed here is very similar to the one we observed in the anthroponotic Leishmania donovani (Laurent et al., 2007), hereby questioning the role of transmission and/or chemotherapeutic drug pressure in the observed population structure.

We hypothesized that Leishmania kDNA may be present in urine of patients with cutaneous leishmaniasis (CL). Urine samples and standard diagnostic specimens were collected from patients with skin lesions. kDNA polymerase chain reaction (PCR) was performed on samples from patients and 10 healthy volunteers from non-endemic areas. Eighty-six of 108 patients were diagnosed with CL and 18 (21%) had detectable Leishmania Viannia kDNA in the urine. Sensitivity and specificity were 20.9% (95% confidence interval [CI] 12.3-29.5%) and 100%. Six of 8 patients with mucocutaneous involvement had detectable kDNA in urine versus 12 of 78 patients with isolated cutaneous disease (P < 0.001). L. (V.) braziliensis (N = 3), L. (V.) guyanensis (N = 6), and L. (V.) peruviana (N = 3) were identified from urine. No healthy volunteer or patient with an alternate diagnosis had detectable kDNA in urine. Sensitivity of urine PCR is sub-optimal for diagnosis. On the basis of these preliminary data in a small number of patients, detectable kDNA in urine may identify less localized forms of infection and inform treatment decisions.

In Argentina, the incidence of American Cutaneous Leishmaniasis (ACL) has shown a steady increase over the last few decades. In the Chaco biogeographical region, specifically, several outbreaks of ACL were recently reported in addition to the usual time-space scattering of ACL cases. However, little is known about the sandfly composition in the eastern, humid Chaco (HC) region or the western, dry Chaco (DC) region. Therefore, phlebotomine captures were performed throughout this region and an analysis of the distribution of reported ACL cases was conducted in order to assess the vector diversity in ACL endemic and epidemic scenarios in the Chaco region. The results support the hypothesis of two distinct patterns: (1) the DC, where Lutzomyia migonei was the most prevalent species, had isolated ACL cases and a zoonotic cycle; (2) the HC, where Lutzomyia neivai was the most prevalent species, had an increase in ACL incidence and outbreaks and an anthropozoonotic cycle. The epidemic risk in the Chaco region may be associated with the current climate trends, landscape modification, connection with other ACL foci, and Lu. neivai predominance and abundance. Therefore, changes in sandfly population diversity and density in the Chaco region are an indicator of emergent epidemic risk in sentinel capture sites., Facultad de Ciencias Naturales y Museo

Epidemiological studies have been conducted to better understand the dynamics of American Cutaneous Leishmaniasis (ACL) in the Amazon region where distinct species of Leishmania circulate. In endemic areas, the optimal diagnosis must be made in the earlier clinical presentation to avoid the complications of chronic disease. The scarcity of financial support, laboratory infrastructure and trained persons are the major obstacles in this reality. This paper describes the result of performing different diagnostic methods for ACL in Amazonas State between the years 2010 and 2011. The tests used were the intradermal skin test (Montenegro's skin test), ELISA (Enzyme-Linked Immunosorbent Assay), direct examination, culture isolation and identification of Leishmania species. A total of 38 suspected human cases of ACL were diagnosed by different methods, of which 71.0% (n = 27) were positive by direct examination, 75.6% (n = 28) had positivity in the culture isolates and, of these, 54.0% (n = 19) had infection with Leishmania (Viannia) guyanensis. The positivity of the intradermal skin test with the leishmanin solution was observed in 77.0% of cases analyzed and the serology with detection of IgG and IgM showed the presence of antibodies in 100% of exams realized results, showing variation in the titles of antibodies. The success of Leishmaniasis treatment depends on an effective and early diagnosis. Parasitological diagnosis is highly specific, but sensitivity is subject to variation because the tissue distribution of parasites generally is not homogeneous and depends on the specie of parasite. Moreover, parasitological tests require invasive procedures and depend on restrictive conditions for the collection of biological sample, which limit their use in large-scale for epidemiological studies. ELISA has been the most widely used serological method for the diagnosis of Visceral Leishmaniasis (VL) as it is easy to perform and has a low cost. However, flaws in specificity are observed in the diagnosis of cutaneous leishmaniasis. Actually the diagnosis needs to be done as an associated methods depending on the question to be solved. © 2016 Elsevier Inc.

Background: Phlebotomine sand flies (Diptera: Psychodidae) are insects of medical importance due to the role that some species play in the transmission of leishmaniasis. This work aimed to study some ecological aspects among sand flies fauna inhabiting two different environments: the várzea (lowland Amazonian forest) and terra firme (upland Amazonian forest), both located in Tefé Municipality, Amazonas State, Braziland to detect Leishmania infection in those phlebotomine populations. Methods: Sand flies were collected using HP light traps. Collection took place over the course of six months: January, February, April, August, September, and October of 2013. To detect natural infection by Leishmania, DNA samples were extracted from female sand flies and submitted to Polymerase Chain Reaction (PCR) targeting the kDNA gene; Leishmania species were identified by PCR-RFLP targeting the hsp70 gene and genetic sequencing. Results: In all, 5,716 individuals were collected, and 46 species were identified. Trichophoromyia ubiquitalis (3,330 - 58.26%) and Nyssomyia antunesi (661 - 11.26%) were the most abundant species. Species richness was greater in terra firme environments (42 species) than in the várzea environments (22 species), and forests ecotopes (43 species) were richer than peridomiciles (28 species). DNA of Leishmania was found in Th. ubiquitalis and Psychodopygus davisi, both of which inhabit the terra firme environment and sequencing analysis confirmed the presence of Leishmania (Viannia) lainsoni DNA in Th. ubiquitalis in Tefé Municipality. Conclusions: The high abundance of Th. ubiquitalis and Ps. davisi and detection of DNA of Leishmania sp. may indicate that both species could be putative vectors for American Cutaneous Leishmaniasis (ACL) in the terra firme environment of Tefé. The sand fly fauna found in várzea is rich and diverse, exhibiting several species, nevertheless the seasonal hydric stress during part of the year that could influence the local diversity, if compared with other studies. This is the first report in Amazonas State of Th. ubiquitalis with presence of L. (V.) lainsoni DNA. © 2015 Pereira et al.; licensee BioMed Central.

The first epidemic tegumentary leishmaniasis ' outbreak in the province of Misiones was recorded in 1998, in the locality of Puerto Esperanza. Phlebotominae collected in the region, previously or simultaneously to the outbreak (September 1993-Decemher 1998) showed that the species Lutzomyia intermedia s. 1. was prevalent (94%, n 6,150) at all the sites sampled with miniature light trap (10) and Shannon trap (3). L. pessoai, L. whitmani, L. migonei, L. shannoni, L. fischeri, L. misionensis, Brumptomyia avellari and B. guimaraesi were also captured. Sand fly distribution in time and space suggests that in the province of Misiones (1) the species already present before 1990 could give rise to the epidemic by the density/dispersion fluctuation of their local populations; (2) the abundance of L. intermedia s. 1. was associated with environments with ecotones of primary-secondary vegetation, close to water bodies and with moderate human disturbance; (3) this species showed, towards the end of 1997, peaks of exceptional abundance, subsequent to rainfall peaks in 1996. This increase in abundance of potential vector sand fly populations close to houses with colonizable surroundings could have generated the 1998 epidemic outbreak., Facultad de Ciencias Naturales y Museo

The authors discuss in this paper the role of inflammatory, anti-inflammatory, and regulatory cytokines in patients infected with different species ofLeishmaniain Amazonas State, Brazil. A comparative analysis was made of serum concentrations of these cytokines in the peripheral blood of 33 patients infected with cutaneous leishmaniasis. The isolates were identified asLeishmania guyanensis,L. naiffi, andL. amazonensis. Most (64%) of the patients were male ranging in age from 18 to 58 years. Protein expression profiles of IL-2, IL-4, IL-6, IL-10, IFN-γ, TNF-α, and IL-17 cytokines were shown to vary significantly between infected and noninfected (control group) individuals and according to theLeishmaniaspecies. Infection caused byL. guyanensisaccounted for 73% of the cases and patients with this parasite also showed higher concentrations of IL-2, IFN-γ, IL-4, and IL-17 when compared to infection byL. amazonensis. Patients with infection caused byL. naiffishowed higher concentration of the cytokines analyzed when compared to uninfected patients; however, there was no statistically significant difference with the other species analyzed.

This study evaluated the pharmacokinetics of topical creams containing 15% paromomycin (“paromomycin alone”) and 15% paromomycin plus 0.5% gentamicin (WR 279,396) in patients with cutaneous leishmaniasis. The investigational creams were applied topically to all lesions once daily for 20 days. Plasma samples were analyzed for simultaneous quantitation of paromomycin and gentamicin isomers and total gentamicin. Pharmacokinetic parameters for gentamicin could not be calculated because detectable levels were rarely evident. After one application, the paromomycin area under the concentration-time curve from 0 to 24 h (AUC 0–24 ) was 2,180 ± 2,621 ng · h/ml (mean ± standard deviation [SD]) for the paromomycin-alone group and 975.6 ± 1,078 ng · h/ml for the WR 279,396 group. After 20 days of application, the paromomycin AUC 0–24 and maximum concentration of drug ( C max ) were 5 to 6 times greater than those on day 1 for both treatment groups. For the paromomycin-alone group, the AUC 0–24 was 8,575 ± 7,268 ng · h/ml and the C max was 1,000 ± 750 ng/ml, compared with 6,037 ± 3,956 ng · h/ml and 660 ± 486 ng/ml for the WR 279,396 group, respectively. Possibly due to large intersubject variability, no differences ( P ≥ 0.05) in the AUC 0–24 or C max were noted between treatment or between sites on day 1 or 20. The percentage of dose absorbed on day 20 was 12.0% ± 6.26% and 9.68% ± 6.05% for paromomycin alone and WR 279,396, respectively. Paromomycin concentrations in plasma after 20 days of application were 5 to 9% of those after intramuscular administration of 15 mg/kg of body weight/day to adults for the systemic treatment of visceral leishmaniasis. Effective topical treatment of cutaneous leishmaniasis appears to be possible with limited paromomycin and gentamicin systemic absorption, thus avoiding drug accumulation and toxicity. (The work described here has been registered at ClinicalTrials.gov under registration no. NCT01032382 and NCT01083576.)

Introduction: This work was carried out on the purpose of identifying the species of phlebotomine sandflies in the municipality of Monte Negro, state of Rondonia, Brazil, that may have been transmitting the American cutaneous leishmaniasis (ACL), and concisely describe epidemiological aspects of disease. METHODS: The epidemiologic and socioeconomical indicators were obtained from government institutions and the local Municipal Secretary of Health. Phlebotomine sandflies were captured using CDC light traps between July 2006 to July 2008. The total of 1,240 of female sandflies were examined by PCR method directed to k-DNA. RESULTS: There has been a significant decrease in the incidence of ACL of about 50% over the last ten years in the municipality. A total of 1,935 specimens of 53 sandfly species were captured, three of the genus Brumptomyia genus and 50 of the genus Lutzomyia. The predominant species was Lutzomyia acanthopharynx, Lutzomyia whitmani, Lutzomyia geniculata and Lutzomyia davisi. None were positive for Leishmania sp. CONCLUSIONS: Four sandflies species were found in the State of Rondonia for the first time: Brumptomyia brumpti, Lutzomyia tarapacaensis, Lutzomyia melloi and Lutzomyia lenti. The presence of Lutzomyia longipalpis, was also captured. Socioeconomical improvement of Brazilian economy and the increase of environmental surveillance in the last 15 years collaborated in the decrease of people exposed to vectors, reducing the incidence of ACL.

Background Traditional methods of detecting Leishmania from cutaneous lesions involve invasive diagnostic procedures, such as scrapings, which cause discomfort, require technical expertise, and carry risks of invasive procedures. We compared the performance of 2 novel, molecular-based non-invasive methods for the diagnosis of cutaneous leishmaniasis (CL). Methods Consecutive patients presenting to the Leishmania Clinic at the Hospital Nacional Cayetano Heredia were enrolled. PCR was performed on filter paper lesion impressions (FPLIs), cytology brushes, and lancets for detection of Leishmania DNA. Smears from lesion scrapings and leishmanin skin test were also performed. Outcome measures were sensitivity and specificity. Composite reference standard was any 2 of 5 tests positive. Species identification was performed by PCR assays of positive specimens. Results Ninety patients with 129 lesions were enrolled, 117 of which fulfilled reference criteria for a diagnosis of CL. Of these 117 lesions, 113 were positive by PCR of lancets used for lesion scrapings versus 116 by PCR of FPLIs (p = 0.930) or 116 by PCR of cytology brushes (p = 0.930). Sensitivity and specificity of PCR on lancets were 96.6% [95% CI 93.3–99.9%] and 100%, respectively. Sensitivity and specificity of FPLI PCR were 99.1% [95% CI 97.4–100%] and 100%, respectively. Sensitivity and specificity of cytology brush PCR were 99.1% [95% CI 97.4–100%] and 100%, respectively. Giemsa-stained lesion smear and leishmanin skin test had inferior sensitivities at 47.9% [95% CI 38.9–57.0%] and 82.3% [95% CI 73.9–90.7%], respectively, compared to PCR of invasive or non-invasive specimens (p

We present an algorithm based on three PCR assays for Leishmania ( Viannia ) species identification and assessed its performance using 70 specimens from Peruvian patients. The succession of the assayed targets can be ordered according to species prevalence. Sequential progression through the algorithm reduced the number of samples here studied by approximately 30% after each step.

The heat-shock protein 70 gene (hsp70) has been exploited for Leishmania species identification in the Old and New World, using polymerase chain reaction (PCR) followed by restriction fragment length polymorphism analysis. Three new Leishmania-specific hsp70 PCRs were recently described, and we applied 2 of these on 89 clinical samples from a total of 73 Peruvian patients with either cutaneous or mucocutaneous leishmaniasis. The new PCRs on average showed a 2- to 3-fold improved sensitivity in the tested sample types (lesion biopsies, aspirates, and scrapings), for both genus detection and species typing, and were most successful in biopsies. Leishmania braziliensis, L. peruviana, and L. guyanensis were encountered. About one third of the L braziliensis parasites contained 2 hsp70 alleles. This study is a paradigm for the implementation of a globally applicable upgraded tool for the identification of Leishmania directly on human specimens from cutaneous and mucocutaneous lesions in the New World. (C) 2012 Elsevier Inc. All rights reserved.

A retrospective analysis was carried out on census data collected from house-to-house surveys during 1991–1992 in 4 areas endemic for Andean cutaneous leishmaniasis (uta) in the Department of Lima, Peru. Major changes in mean annual incidence in susceptible persons have taken place in these sites during the last 60 years. In particular, there is strong support for the hypothesis that, from the 1950s to the 1970s, the transmission rate was temporarily suppressed, largely as a by-product of the DDT house spraying campaign against malaria. These results are consistent with (i) anecdotal evidence, contemporary with the spraying campaign, and (ii) the official Ministry of Health records for the annual number of uta cases in the Departments of Lima and Ancash.

International audience; This review gives an update of current knowledge on the clinical pleiomorphism of Leishmania, with a special emphasis on the case of asymptomatic carriage. The first part describes the numerous unusual expressions of the disease that occur besides the classic (visceral, cutaneous, and mucocutaneous) forms of leishmaniases. The second part deals with progress in the understanding of disease outcome in humans, and the possible future approaches to improve our knowledge in the field. The third part highlights the role of the too often neglected asymptomatic carrier compartment. This group could be key to understanding infraspecific differences in virulence and pathogenicity of the parasite, as well as identifying the genetic determinants involved in the expression of the disease.

We report a case of a 9-year-old boy who presented with 2 lesions that were compatible clinically with cutaneous leishmaniasis of the New World. A skin biopsy showed tuberculoid granulomas with rounded calcified bodies. The diagnosis was supported by a positive leishmanin test and a positive polymerase chain reaction. The patient responded to specific treatment for leishmaniasis. These calcified bodies, described under different names in the literature (Michaelis-Gutmann bodies, Schaumann bodies, psammoma bodies, or conchoidal bodies), have been reported in an experimental leishmaniasis and, only once before, in a case of human leishmaniasis of the Old World.

We compared traditional cutaneous leishmaniasis diagnostic methods to filter paper lesion impression (FPLI) PCR for secondarily infected ulcers and nonulcerative lesions. The sensitivity and specificity of FPLI PCR for secondarily infected lesions ( n = 8) were 100%. In primarily nonulcerative lesions ( n = 15), the sensitivity of FPLI PCR was inferior to that of pooled-invasive-specimen PCR (72.7% versus 100%) ( P = 0.10). FPLI PCR is sensitive, specific, and unlike invasive procedures, can be used in secondarily infected ulcers. Invasive specimen collection is superior in nonulcerative lesions.

Background: Traditional methods of diagnosing mucosal leishmaniasis (ML), such as biopsy with histopathology, are insensitive and require collection of an invasive diagnostic specimen. Methods: We compared standard invasive procedures including biopsy histopathology, biopsy PCR, and leishmanin skin test (LST) to a novel, non-invasive, cytology-brush based PCR for the diagnosis of ML in Lima, Peru. Consensus reference standard was 2/4 tests positive, and outcome measures were sensitivity and specificity. Leishmania species identification was performed by PCR-based assays of positive specimens. Results: Twenty-eight patients were enrolled, 23 of whom fulfilled criteria for a diagnosis of ML. Sensitivity and specificity of biopsy with histopathology were 21.7% [95% CI 4.9-38.5%] and 100%; 69.6% [95% CI 50.8-88.4%] and 100% for LST; 95.7% [95% CI 87.4-100%] and 100% for biopsy PCR; and 95.7% [95% CI 87.4-100%] and 90% [95% CI 71.4-100%] for cytology brush PCR using both Cervisoft® and Histobrush® cervical cytology brushes. Represented species identified by PCR-RFLP included: L. (V). braziliensis (n = 4), and L. (V). peruviana (n = 3). Conclusions: Use of commercial grade cytology brush PCR for diagnosis of ML is sensitive, rapid, well tolerated, and carries none of the risks of invasive diagnostic procedures such as biopsy. Further optimization is required for adequate species identification. Further evaluation of this method in field and other settings is warranted.

Background Increased rates for failure in leishmaniasis antimony treatment have been recently recognized worldwide. Although several risk factors have been identified there is no clinical score to predict antimony therapy failure of cutaneous leishmaniasis. Methods A case control study was conducted in Peru from 2001 to 2004. 171 patients were treated with pentavalent antimony and followed up to at least 6 months to determine cure or failure. Only patients with ulcerative cutaneous leishmaniasis (N = 87) were considered for data analysis. Epidemiological, demographical, clinical and laboratory data were analyzed to identify risk factors for treatment failure. Two prognostic scores for antimonial treatment failure were tested for sensitivity and specificity to predict antimony therapy failure by comparison with treatment outcome. Results Among 87 antimony-treated patients, 18 (21%) failed the treatment and 69 (79%) were cured. A novel risk factor for treatment failure was identified: presence of concomitant distant lesions. Patients presenting concomitant-distant lesions showed a 30.5-fold increase in the risk of treatment failure compared to other patients. The best prognostic score for antimonial treatment failure showed a sensitivity of 77.78% and specificity of 95.52% to predict antimony therapy failure. Conclusions A prognostic score including a novel risk factor was able to predict antimonial treatment failure in cutaneous leishmaniasis with high specificity and sensitivity. This prognostic score presents practical advantages as it relies on clinical and epidemiological characteristics, easily obtained by physicians or health workers, and makes it a promising clinical tool that needs to be validated before their use for developing countries., Author Summary The manuscript is relevant because of the finding of a new risk factor for chemotherapy failure and the development of a prognosis score for cutaneous leishmaniasis. The proportion of patients that have multiple lesions in American Tegumentary Leishmaniasis (ATL) is considerable. Publications and our experience permit to estimate that they represent around 20% of the affected population from the Amazon basin with cutaneous lesions. In addition, about 1/3 of them would correspond to the concomitant distant lesions category, the novel risk factor identified with a very high odds ratio (20–30) associated. Such numbers merit study of concomitant distant ulcers category on its own, not only because of clinical management implications, but also to search for factors that are contributing to chemotherapy failure. Finally, the simple equation proposed in the manuscript can be easily adapted to smart phone technologies. Similar prognosis equations are scarce for other pathologies and do not exist for Cutaneous Leishmaniasis at all. The simplicity of this tool should be followed by subsequent epidemiologic studies in other ATL endemic regions.

In much of the endemic area for cutaneous leishmaniasis (uta) in the Peruvian Andes, the only 2 anthropophilic sandfly species present are Lutzomyia peruensis and Lu. verrucarum. On the basis of a single confirmed isolation of Leishmania peruviana (the aetiological agent of uta) from a wild Lu. peruensis, and apparent associations between sandfly abundance and the incidence of uta, it is generally believed that Lu. peruensis is the most important vector. In this paper, a potential role for Lu. verrucarum in the transmission of uta is indicated by laboratory experiments which show that this species is vectorially competent for L. peruviana. Individual or pooled colonized sandflies were permitted to take a second blood meal on 22 susceptible golden hamsters at varying intervals after feeding on hamsters previously infected with L. peruviana. Transmission was achieved by a single infected sandfly (of a total of 59) following a 15 d incubation period. Transmission was recognized by the characteristic clinical response (footpad swelling) associated with hamsters which have been inoculated with L. peruviana, and by the presence of parasites in aspirates made from the swollen footpad, detected by the polymerase chain reaction (PCR) and by parasite isolations in biphasic blood-agar culture medium. The identity of the parasite isolates was also confirmed by PCR (specific for parasites in the L. braziliensis complex). This is the first reported experimental transmission of L. peruviana by any sandfly species.

In Perd tegumentary leishmaniases are endemic, more than 5000 new cases per year being reported by the Ministry of Health, although the estimated real incidence is three times as great, and human immunodeficiency virus (HIV) infections and the reported incidence of the acquired immune deficiency syndrome (AIDS) are increasing. The more frequently reported infectious diseases associated with AIDS patients in Perd are diarrhoea due to coccidia, extrapulmonary tuberculosis, cerebral toxoplasmosis, and cryptococcal meningitis...

Background American cutaneous leishmaniasis (ACL) is a re-emerging disease in the state of São Paulo, Brazil. It is important to understand both the vector and disease distribution to help design control strategies. As an initial step in applying geographic information systems (GIS) and remote sensing (RS) tools to map disease-risk, the objectives of the present work were to: (i) produce a single database of species distributions of the sand fly vectors in the state of São Paulo, (ii) create combined distributional maps of both the incidence of ACL and its sand fly vectors, and (iii) thereby provide individual municipalities with a source of reference material for work carried out in their area. Results A database containing 910 individual records of sand fly occurrence in the state of São Paulo, from 37 different sources, was compiled. These records date from between 1943 to 2009, and describe the presence of at least one of the six incriminated or suspected sand fly vector species in 183/645 (28.4%) municipalities. For the remaining 462 (71.6%) municipalities, we were unable to locate records of any of the six incriminated or suspected sand fly vector species (Nyssomyia intermedia, N. neivai, N. whitmani, Pintomyia fischeri, P. pessoai and Migonemyia migonei). The distribution of each of the six incriminated or suspected vector species of ACL in the state of São Paulo were individually mapped and overlaid on the incidence of ACL for the period 1993 to 1995 and 1998 to 2007. Overall, the maps reveal that the six sand fly vector species analyzed have unique and heterogeneous, although often overlapping, distributions. Several sand fly species - Nyssomyia intermedia and N. neivai - are highly localized, while the other sand fly species - N. whitmani, M. migonei, P. fischeri and P. pessoai - are much more broadly distributed. ACL has been reported in 160/183 (87.4%) of the municipalities with records for at least one of the six incriminated or suspected sand fly vector species, while there are no records of any of these sand fly species in 318/478 (66.5%) municipalities with ACL. Conclusions The maps produced in this work provide basic data on the distribution of the six incriminated or suspected sand fly vectors of ACL in the state of São Paulo, and highlight the complex and geographically heterogeneous pattern of ACL transmission in the region. Further studies are required to clarify the role of each of the six suspected sand fly vector species in different regions of the state of São Paulo, especially in the majority of municipalities where ACL is present but sand fly vectors have not yet been identified.

Leishmania naiffi was isolated from 10 out of 64 armadillos (Dasypus novemcinctus) examined in Amazonas, Pará and Rondônia States in the Brazilian Amazon Region. The isolates were obtained in culture from samples of liver (3), spleen (3), lymph nodes (2), skin (1) and blood (1) from the infected animals. Heavy infections with the same parasite were detected for the first time in Psychodopygus squamiventris, a common man-biting phlebotomine, in Amazonas and Pará. A new case of cutaneous leishmaniasis caused by L. naiffi is described from the Manaus area, making a total of three known cases of human infection by this parasite.