Human Visceral Leishmaniasis Research Articles

Detection of Leishmania spp. in Chronic Dermatitis: Retrospective Study in Exposed Horse Populations.

Leishmania infantum is a protozoan causing human zoonotic visceral leishmaniasis (ZVL) and visceral–cutaneous canine leishmaniosis (CanL) in the Mediterranean Basin. L. infantum is able to infect a large number of wild and domestic species, including cats, dogs, and horses. Since the 1990s, clinical cases of equine leishmaniasis (EL), typically characterized by cutaneous forms, have been increasingly diagnosed worldwide. The aim of the present study was to evaluate the presence of clinical forms of EL in CanL-endemic areas in Italy, where exposure of equine populations was ascertained from recent serological surveys. For this purpose, formalin-fixed and paraffin-embedded skin biopsies of 47 horses presenting chronic dermatitis compatible with EL were retrospectively selected for the study and subjected to conventional and q-PCR. A singular positivity for L. infantum was found; BLAST analysis of sequence amplicons revealed a 99–100% homology with L. infantum sequences. The histological examination revealed a nodular lymphoplasmacytic and histiocytic infiltrate; immunohistochemistry showed rare macrophages containing numerous positive amastigotes. The present retrospective study reports, for the first time, a case of a cutaneous lesion by L. infantum occurring in an Italian horse. Pathological and healthy skin samples should be investigated on a larger scale to provide information on the potential clinical impact of EL in the practice, and to define the role of horses in epidemiological ZVL and CanL scenarios.

The current epidemiology of leishmaniasis in Turkey, Azerbaijan and Georgia and implications for disease emergence in European countries

Leishmania spp. are sand fly‐borne protozoan parasites causing leishmaniasis in humans and animals. The aim of the study was to analyse the epidemiology of leishmaniasis in Turkey, Azerbaijan and Georgia from 2005 to 2020 and evaluate the associated risk for disease emergence in European countries. It is based on an analysis of WHO and OIE reported cases between 2005 and 2020, a review of scientific articles published in SCOPUS between 2009 and 2020 and a questionnaire survey to public health and veterinary authorities in these countries. Endemic Leishmania spp. include L. infantum in the three countries, L. major in Azerbaijan and Turkey and L. tropica and L. donovani in Turkey. Leishmaniasis is reported in humans, animals and sand flies and incidence is spatially and temporarily variable. In the southern Caucasus and particularly in Georgia, reported incidence of human visceral leishmaniasis by L. infantum remains high. However, whilst Georgia experienced a gradual decrease from >4.0 cases per 100,000 population in 2005–09 to 1.13 cases per 100,000 population in 2020, the period with highest incidence in Azerbaijan, which ranged between 0.40 and 0.61 cases per 100,000 population, was 2016–2019, and no cases have so far been reported for 2020. Visceral leishmaniasis in the Southern Caucasus affects mostly young children from deprived urban areas and its closely associated to canine leishmaniasis. Turkey reported cases of visceral leishmaniasis between 2005 and 2012 and in 2016 only, and incidence ranged between 0.02 and 0.05 per 100,000 population. In contrast, the reported annual incidence of cutaneous leishmaniasis in Turkey was much greater and peaked at 7.02 cases per 100,000 population in 2013, associated to imported cases from cutaneous leishmaniasis endemic Syria. Leishmaniasis by L. infantum in Azerbaijan and Georgia represents a regional public and animal health challenge that requires support to improve diagnosis, treatment and control. The unprecedented rise of cutaneous leishmaniasis and the spread of L. tropica and L. donovani in Turkey is an important risk factor for their emergence in Europe, especially in Mediterranean countries where competent vectors are widespread.

PREVALENCE OF VISCERAL LEISHMANIASIS AMONG ADULTS IN SANA’A CITY-YEMEN

Background and Aims: Visceral leishmaniasis (VL) is a zoonotic and human disease caused by species of Leishmania. Parasites are transmitted to the vertebrate host by the bite of a sand fly female (Phlebotomus longipalpis), where the infected promastigotes transform into amastigotes; and this is deadly if left untreated. The purposes of the current research were to reveal the prevalence and potential risk factors for VL in adults in Sana’a city, Yemen. Subjects and methods: This cross-sectional study was performed during the period from January 2020 to November 2020. Individuals who come for a regular medical examination at Al-Zahrawi Medical Center, Althobhani Specialist Laboratories, Police Hospital, and University of Science and Technology Hospital. A target sample size of 300 was selected, and serum samples were collected from all subjects to determine the prevalence of anti-VL antibodies in human by immune-chromatographic assay using K39 recombinant antigen. Results: The ages of the participants' ranged from 18-65 years, with a mean of 29.8±8.2 years. The positive rate of antibodies against Leishmania species by immune-chromatographic dipstick strip (rK39) was 6.0%. There was statistically important association linking male gender and contracting VL (8.8%, OR=4.1, CI=1.2-14.4, P=0.01). There was a significant association (<0.001) between the presence of dogs, rats, and goats in or around live houses and positive VL antibodies with an OR equal to 8.8.7.3 and 8.4, respectively. There were significant risk factors for garbage around the living house, there was also a significant association between displacement and the incidence of VL (P<0.001) (OR=8.6, CI=2.8–27.2). Conclusion: Visceral leishmaniasis was highly prevalent in Sana'a city, and potential risk factors for VL were present with displacement, dogs, rats, goats, garbage, sleeping outside enclosed rooms, and sand flies in living houses. Further studies of human VL need to be conducted to clarify this issue in Yemen, to track and confirm potential reservoirs among canines and other animals, as well as to study vectors. Keywords: adults, immune-chromatographic assay, prevalence, potential risk factors, recombinant antigen K39, Sana’a city, visceral leishmaniasis, Yemen. Peer Review History: Received: 9 March 2022; Revised: 14 April; Accepted: 29 April, Available online: 15 May 2022 Academic Editor: Dr. Sally A. El-Zahaby, Pharos University in Alexandria, Egypt, sally.elzahaby@yahoo.com UJPR follows the most transparent and toughest ‘Advanced OPEN peer review’ system. The identity of the authors and, reviewers will be known to each other. This transparent process will help to eradicate any possible malicious/purposeful interference by any person (publishing staff, reviewer, editor, author, etc) during peer review. As a result of this unique system, all reviewers will get their due recognition and respect, once their names are published in the papers. We expect that, by publishing peer review reports with published papers, will be helpful to many authors for drafting their article according to the specifications. Auhors will remove any error of their article and they will improve their article(s) according to the previous reports displayed with published article(s). The main purpose of it is ‘to improve the quality of a candidate manuscript’. Our reviewers check the ‘strength and weakness of a manuscript honestly’. There will increase in the perfection, and transparency. Received file: Reviewer's Comments: Average Peer review marks at initial stage: 6.0/10 Average Peer review marks at publication stage: 7.0/10 Reviewers: Dr. Tamer Elhabibi, Suez Canal University, Egypt, tamer_hassan@pharm.suez.edu.eg Dr. Nicola Micale, University of Messina, Italy, nmicale@unime.it Dr. Rawaa Souhil Al-Kayali, Aleppo University, Syria, rawah67@hotmail.com Similar Articles: PREVALENCE OF HEPATITIS G VIRUS AMONG PATIENTS WITH CHRONIC LIVER DISEASE AND HEALTHY INDIVIDUALS, SANA'A CITY-YEMEN

T Lymphocyte Exhaustion During Human and Experimental Visceral Leishmaniasis.

A key point of immunity against protozoan Leishmania parasites is the development of an optimal T cell response, which includes a low apoptotic rate, high proliferative activity and polyfunctionality. During acute infection, antigen-specific T cells recognize the pathogen resulting in pathogen control but not elimination, promoting the development and the maintenance of a population of circulating effector cells that mount rapid response quickly after re-exposure to the parasite. However, in the case of visceral disease, the functionality of specific T cells is lost during chronic infection, resulting in inferior effector functions, poor response to specific restimulation, and suboptimal homeostatic proliferation, a term referred to as T cell exhaustion. Multiple factors, including parasite load, infection duration and host immunity, affect T lymphocyte exhaustion. These factors contribute to antigen persistence by promoting inhibitory receptor expression and sustained production of soluble mediators, influencing suppressive cell function and the release of endogenous molecules into chronically inflamed tissue. Together, these signals encourage several changes, reprogramming cells into a quiescent state, which reflects disease progression to more severe forms, and development of acquired resistance to conventional drugs to treat the disease. These points are discussed in this review.

MiRNA-21 regulates CD69 and IL-10 expression in canine leishmaniasis.

Visceral leishmaniasis in humans is a chronic and fatal disease if left untreated. Canine leishmaniasis (CanL) is a severe public health problem because infected animals are powerful transmitters of the parasite to humans via phlebotomine vectors. Therefore, dogs are an essential target for control measures. Progression of canine infection is accompanied by failure of cellular immunity with reduction of circulating lymphocytes and increased cytokines that suppress macrophage function. Studies showed that the regulation of the effector function of macrophages and T cells appears to depend on miRNAs; miRNA-21 (miR-21) shows increased expression in splenic leukocytes of dogs with CanL and targets genes related to the immune response. Mimics and inhibitors of miR-21 were used in vitro to transfect splenic leukocytes from dogs with CanL. After transfection, expression levels of the proteins FAS, FASL, CD69, CCR7, TNF-α, IL-17, IFN-γ, and IL-10 were measured. FAS, FASL, CD69, and CCR7 expression levels decreased in splenic leukocytes from dogs with CanL. The miR-21 mimic decreased CD69 expression in splenic leukocytes from CanL and healthy groups. The miR-21 inhibitor decreased IL-10 levels in culture supernatants from splenic leukocytes in the CanL group. These findings suggest that miR-21 alters the immune response in CanL; therefore, miR-21 could be used as a possible therapeutic target for CanL.

Experimental evolution links post-transcriptional regulation to Leishmania fitness gain.

The protozoan parasite Leishmania donovani causes fatal human visceral leishmaniasis in absence of treatment. Genome instability has been recognized as a driver in Leishmania fitness gain in response to environmental change or chemotherapy. How genome instability generates beneficial phenotypes despite potential deleterious gene dosage effects is unknown. Here we address this important open question applying experimental evolution and integrative systems approaches on parasites adapting to in vitro culture. Phenotypic analyses of parasites from early and late stages of culture adaptation revealed an important fitness tradeoff, with selection for accelerated growth in promastigote culture (fitness gain) impairing infectivity (fitness costs). Comparative genomics, transcriptomics and proteomics analyses revealed a complex regulatory network associated with parasite fitness gain, with genome instability causing highly reproducible, gene dosage-independent and -dependent changes. Reduction of flagellar transcripts and increase in coding and non-coding RNAs implicated in ribosomal biogenesis and protein translation were not correlated to dosage changes of the corresponding genes, revealing a gene dosage-independent, post-transcriptional mechanism of regulation. In contrast, abundance of gene products implicated in post-transcriptional regulation itself correlated to corresponding gene dosage changes. Thus, RNA abundance during parasite adaptation is controled by direct and indirect gene dosage changes. We correlated differential expression of small nucleolar RNAs (snoRNAs) with changes in rRNA modification, providing first evidence that Leishmania fitness gain in culture may be controlled by post-transcriptional and epitranscriptomic regulation. Our findings propose a novel model for Leishmania fitness gain in culture, where differential regulation of mRNA stability and the generation of modified ribosomes may potentially filter deleterious from beneficial gene dosage effects and provide proteomic robustness to genetically heterogenous, adapting parasite populations. This model challenges the current, genome-centric approach to Leishmania epidemiology and identifies the Leishmania transcriptome and non-coding small RNome as potential novel sources for the discovery of biomarkers that may be associated with parasite phenotypic adaptation in clinical settings.

Bayesian latent class models for identifying canine visceral leishmaniosis using diagnostic tests in the absence of a gold standard.

Like many infectious diseases, there is no practical gold standard for diagnosing clinical visceral leishmaniasis (VL). Latent class modeling has been proposed to estimate a latent gold standard for identifying disease. These proposed models for VL have leveraged information from diagnostic tests with dichotomous serological and PCR assays, but have not employed continuous diagnostic test information. In this paper, we employ Bayesian latent class models to improve the identification of canine visceral leishmaniasis using the dichotomous PCR assay and the Dual Path Platform (DPP) serology test. The DPP test has historically been used as a dichotomous assay, but can also yield numerical information via the DPP reader. Using data collected from a cohort of hunting dogs across the United States, which were identified as having either negative or symptomatic disease, we evaluate the impact of including numerical DPP reader information as a proxy for immune response. We find that inclusion of DPP reader information allows us to illustrate changes in immune response as a function of age. Utilization of continuous DPP reader information can improve the correct discrimination between individuals that are negative for disease and those with clinical VL. These models provide a promising avenue for diagnostic testing in contexts with multiple, imperfect diagnostic tests. Specifically, they can easily be applied to human visceral leishmaniasis when diagnostic test results are available. Also, appropriate diagnosis of canine visceral leishmaniasis has important consequences for curtailing spread of disease to humans.

Down regulation of IL-10 and TGF-β1 mRNA expression associated with reduced inflammatory process correlates with control of parasitism in the liver after treating L. infantum infected dogs with the LBMPL vaccine therapy

The liver plays an important role in human and canine visceral leishmaniasis, then it is considered as target to understand the mechanisms involved in the parasite control and a parameter to assess therapeutic responses. In this sense, our study focuses on evaluating the major alterations in the liver by histological (morphometric parenchyma inflammation/semi-quantitative portal inflammation), immunohistochemical assays (parasitism), and qPCR (parasitism and cytokine gene expression) in Leishmania infantum naturally infected dogs and treated with LBMPL vaccine. Animals were divided in four groups: NI group (n = 5): uninfected and untreated dogs; INT group (n = 7): L. infantum-infected dogs and not treated; MPL group (n = 6): L. infantum-infected dogs that received only monophosphoryl lipid A adjuvant, and LBMPL group (n = 10): L. infantum-infected dogs that received treatment with the vaccine composed by L. braziliensis disrupted promastigotes associated with MPL adjuvant. Ninety days after the end of treatments, the dogs were euthanized, and the liver was collected for the proposed evaluations. Significantly lower portal inflammatory reactions, and lower parenchyma inflammation were observed in the LBMPL group compared to INT and MPL groups. iNOS mRNA expression was higher in LBMPL group and in contrast, IL-10 and TGF-β1 mRNA expression was lower in this group when compared to INT group. Immunohistochemical and qPCR analysis showed significant parasite load reduction in LBMPL group compared to INT and MPL animals. Our data suggest that in naturally Leishmania-infected dogs, LBMPL vaccine reduces the damage in the hepatic tissue, being able to attenuate the type 2 immune response. It could be associated with a marked reduction in the parasitism decreasing liver inflammation in treated dogs. Along with previously obtained data, our results suggest that LBMPL vaccine can significantly contribute to the therapy strategy for L. infantum infected dogs.

MiR-548d-3pIs Up-Regulated in Human Visceral Leishmaniasis and Suppresses Parasite Growth inMacrophages.

Visceral leishmaniasis caused by Leishmania (Leishmania) infantum in Latin America progress with hepatosplenomegaly, pancytopenia, hypergammaglobulinemia, and weight loss and maybe lethal mainly in untreated cases. miRNAs are important regulators of immune and inflammatory gene expression, but their mechanisms of action and their relationship to pathogenesis in leishmaniasis are not well understood. In the present study, we sought to quantify changes in miRNAs associated with immune and inflammatory pathways using the L. (L.) infantum promastigote infected- human monocytic THP-1 cell model and plasma from patients with visceral leishmaniasis. We identified differentially expressed miRNAs in infected THP-1 cells compared with non-infected cells using qPCR arrays. These miRNAs were submitted to in silico analysis, revealing targets within functional pathways associated with TGF-β, chemokines, glucose metabolism, inflammation, apoptosis, and cell signaling. In parallel, we identified differentially expressed miRNAs in active visceral leishmaniasis patient plasma compared with endemic healthy controls. In silico analysis of these data indicated different predicted targets within the TGF-β, TLR4, IGF-I, chemokine, and HIF1α pathways. Only a small number of miRNAs were commonly identified in these two datasets, notably with miR-548d-3p being up-regulated in both conditions. To evaluate the potential biological role of miR-548d-3p, we transiently transfected a miR-548d-3p inhibitor into L. (L.) infantum infected-THP-1 cells, finding that inhibition of miR-548d-3p enhanced parasite growth, likely mediated through reduced levels of MCP-1/CCL2 and nitric oxide production. Further work will be required to determine how miR-548d-3p plays a role in vivo and whether it serves as a potential biomarker of progressive leishmaniasis.

Revealing a Novel Antigen Repressor of Differentiation Kinase 2 for Diagnosis of Human Visceral Leishmaniasis in India.

Visceral leishmaniasis (VL) is one of the major global health concerns due to its association with morbidity and mortality. All available diagnostic tools have been, until now, unable to provide a very specific and cost-effective mode of detection for VL globally. Therefore, the design of robust, specific, and commercially translatable diagnostic tests is urgently required. Currently, we are attempting to identify and explore the diagnostic potential of a novel parasite antigen. Repressor of differentiation kinase 2 (RDK2), a serine/threonine kinase, has a versatile role in parasite life cycle progression. However, its role as a diagnostic candidate for VL has not been investigated. Herein, we cloned and over-expressed LdRDK2 and studied the recombinant RDK2 for the diagnosis of human VL using serum and urine samples. In silico analysis predicted that RDK2 is conserved among Leishmania species with the least conservation in humans. RDK2 developed immune-reactive bands with antibodies present in VL patients’ sera, and it demonstrated no cross-reactivity with sera from healthy controls and other diseases. Additionally, RDK2 antigen demonstrated a significant reactivity with IgG antibodies of VL patients’ sera, with 78% sensitivity and 86.67% specificity as compared to healthy controls and other diseases. Furthermore, we evaluated its utility for non-invasive diagnosis of VL using patients’ urine samples and found 93.8% sensitivity and 85.7% specificity. RDK2 was found to have better sensitivity and treatment response in patients’ urine compared to serum samples, indicating its role as a promising point of care (POC) antigen. In a nutshell, we explored the role of RDK2 as a potential diagnostic marker for VL in both invasive and non-invasive modes as well as its utility as a promising POC antigen for treatment response cases.

Epidemiological Aspects and High Magnitude of Human Visceral Leishmaniasis in Ceará, Northeast of Brazil, 2007-2021.

ABSTRACTBackground: Human visceral leishmaniasis HVL is endemic to 75 countries. The state of Ceará, the Northeast region of Brazil, is of great sanitary importance for the transmission of HVL, and it stands out as an area of interest for epidemiological aspects and control strategies. This study aimed to characterize HVL in relation to epidemiological aspects, composite incidence, and mortality rates in the state of Ceará, Brazil, from 2007 to 2021. Methods: This ecological study used temporal and spatial cuts of HVL data from the notifiable diseases information system. Epidemiological indicators such as incidence, mortality, and composite indices of incidence and mortality were calculated according to the Ministry of Health standardization. Results: There were 6,775 confirmed cases, with high incidence coefficients in 2009 6.96 cases/100,000 inhabitants and 2011 9.83 cases/100,000 inhabitants, and the highest mortality rate in 2011 6.96 deaths/100,000 inhabitants. The composite index of incidence and mortality identified municipalities in the Northern, Northwestern, and Southern regions of Ceará as having the highest risk of HVL. Conclusions: HVL remained endemic throughout the study period, with epidemiological indicators and risk of transmission expressing high magnitude, mainly in the Northeast, Northwest, and South regions of Ceará.

Epidemiological and clinical factors associated with lethality from Human Visceral Leishmaniasis in Northeastern Brazil, 2007 to 2018.

ABSTRACTHuman Visceral Leishmaniasis (HVL) presents a subacute clinical evolution with systemic involvement, which can result in high case fatality, especially among untreated individuals or those with low socioeconomic status. This study aimed to identify epidemiological and clinical factors associated with HVL case fatality in the Ceara State, from 2007 to 2018. This is an analytical cross-sectional study. The bivariate analysis was performed by Stata 15.1 using Pearson’s Chi-square or Fisher’s exact test; and Poisson regression for age-controlled multivariate analysis. From 2007 to 2018, there were 4,863 new confirmed cases and 343 deaths from HVL (case fatality rate=7.05%). The risk factors associated with case fatalities were: age group (RR=8.69; 95%CI:3.56-21.20); black population (RR=2.21; 95%CI:1.45-3.35); jaundice symptoms (RR=1.72; 95%CI:1.38-2.14); edema (RR=2.62; 95%CI:2.10-3.26) and hemorrhagic phenomena (RR=1.63; 95%CI:1.26-2.10); and no prescription drug intake (RR=4.03; 95%CI:2.98-5.46). Treatment with pentavalent antimonial was a protective factor (RR=0.35; 95%CI:0.27-0.45). The number of deaths increased among the elderly, illiterate, urban residents, and black skin color individuals. The drugs pentavalent antimonial and amphotericin B showed an association with death, but were not considered causal factors. Treatment failure led to a high risk of death. In multivariate analysis, the risk factors for fatal cases were age group, black skin, symptoms of jaundice, edema and hemorrhagic phenomena; and failure to take the prescription drugs. Treatment with pentavalent antimonial was shown to be a protective factor. Knowing the factors associated with the fatality of VL-HIV cases may help to improve public policies, in order to refine the epidemiological surveillance program and, consequently, prevent deaths related to the disease in Ceara.

Culicoides' species from a Leishmania transmission hotspot and efficacy of the Captor® suction light trap.

Culicoides have medical and veterinary importance, as they play a role as vectors of viruses, protozoa, and nematodes that cause diseases. Despite the relevance to public health, greater attention has been given to other insect vectors. Therefore, the objective of this study was to evaluate the efficiency of the Captor® light trap in capturing Culicoides that could be examined for the presence of Leishmania DNA. The insects were captured in a rural area of Santa Maria, Rio Grande do Sul, Brazil, where canine and human visceral leishmaniasis have been diagnosed. Adult insects were collected weekly, from 6:00 pm to 6:00 am, for a 12 month period using a Captor® brand suction light trap. All Culicoides were identified at species level. Pools of Culicoides were tested using the Polymerase Chain Reaction (PCR) technique for the detection of Leishmania DNA. A total of 16,016 specimens were collected (71.54% females and 28.39% males), divided among seven species. In the DNA analysis, none of the pools showed a positive result for Leishmania. The data presented demonstrate that the trap is efficient and can be an alternative for use in entomological research. They also demonstrate that, despite the females having hematophagous habits, similar to other vector insects, they did not have contact with Leishmania in the studied locality.

CRYOPRESERVATION OF LEISHMANIA DONOVANI PROMASTIGOTES AT -80°C

Visceral leishmaniasis (VL) is one of the important parasitic diseases caused by the obligate intracellular protozoan parasite of Leishmania donovani complex. The disease is associated with severe morbidity and is fatal if left untreated. Human VL is rare in Bulgaria, but local cases are registered annually in the country. Diagnosis of the disease is based on clinical and epidemiological data and the different laboratory methods for ethiological diagnosis. Isolation of the causative agent through cultural methods, allows to make an extremely accurate diagnosis, but require time and cost of materials. Cryopreservation of isolated parasites allows their keeping for a long time and recovery in need of subsequent immunological and molecular analyzes. The described and optimized cryopreservation method of Leishmania species promastigotes in laboratory conditions is easily feasible and reliable and can be used for long-term storage of these parasites without changing their viability and biological characteristics.

Spatial and spatiotemporal patterns of human visceral leishmaniasis in an endemic southeastern area in countryside Brazil.

ABSTRACTVisceral leishmaniasis (VL) has shown endemic pattern and epidemic episodes in urban and rural areas, however, there are still gaps in knowledge with regards to disease transmission. This study aimed to analyze the spatiotemporal dispersion of VL cases in the municipality of Araçuaí, Minas Gerais. A study of confirmed VL cases was conducted considering the endemic and epidemic periods between 2012 and 2017. The incidence rate was calculated, and for spatial analysis, the kernel map, directional distribution ellipse, and space-time scanning techniques were used. The correlations between VL cases and exposure variables (precipitation, humidity, and temperature) were calculated. The mean incidence of VL in the endemic period was 18.5 (95% confidence interval (CI) 5.9-32.5) and 44.4 in the epidemic period (95%CI, 12.0-28.6) by 100,000 inhabitants. The relative risk for the epidemic period was 2.4 (95% CI 1.4-4.1) when compared to the endemic period. A higher incidence of the disease was observed in rural areas of the municipality. Kernel mapping analysis revealed hotspots in the urban area of the municipality. The directional distribution ellipse encompasses the urban perimeter and part of the rural area of the municipality, expanding eastward during the epidemic period. Spatial analysis revealed a high-risk cluster for VL in rural areas. A positive correlation was observed between VL cases and temperature during the endemic period. Spatial analysis allowed us to outline the epidemiological scenario of human VL cases. These findings may be useful in case surveillance and in the work of health professionals and managers in Brazil.

Prevalence of Leishmania infantum in Dogs from Deforested Areas of the Amazon Biome.

Background and Objectives: Species of Leishmania genus are intracellular parasites responsible for severe zoonotic diseases worldwide, such as leishmaniasis. In Brazil, the most important species is Leishmania infantum. In the northern region, the state of Pará is notable, with a high number of cases reported in recent years. The second largest number of cases of human visceral leishmaniasis in the region in recent years has been reported in the city of Marabá (Pará state). We investigated the prevalence of L. infantum in domestic dogs from Marabá in Amazon region in Brazil. Materials and Methods: A total of 400 blood samples were tested using the dual-path platform chromatographic immunoassay (DPP® CVL-Bio-Manguinhos), and molecular diagnosis based in cathepsin L-like gene, which has better specificity and sensitivity than other similar tests. Results: The prevalence of L. infantum was 75.5% (302/400) in the serological test, and 59.25% (237/400) in molecular diagnosis, and 45.5% (182/400) in both tests. Interpretation and Conclusion: The results confirmed that the parasite L. infantum was prevalent in the dog population of the studied region and adoption of appropriate public policies are urgently needed.

Canine leishmaniosis in Tunisia: Growing prevalence, larger zones of infection.

BackgroundDiscovered by Nicolle and Comte in 1908 in Tunisia, Leishmania infantum is an intracellular protozoan responsible for zoonotic canine leishmaniosis (CanL) and zoonotic human visceral leishmaniasis (HVL). It is endemic in several regions of the world, including Tunisia, with dogs considered as the main domestic reservoir. The geographic expansion of canine leishmaniosis (CanL) has been linked to global environmental changes that have affected the density and the distribution of its sand fly vectors.Methodology/Principal findingsIn this study, a cross-sectional epidemiological survey on CanL was carried out in 8 localities in 8 bioclimatic areas of Tunisia. Blood samples were taken from 317 dogs after clinical examination. Collected sera were tested by indirect fluorescent antibody test (IFAT; 1:80) for the presence of anti-Leishmania infantum antibodies. The overall seroprevalence was 58.3% (185/317). Among positive dogs, only 16.7% showed clinical signs suggestive of leishmaniosis. Seroprevalence rates varied from 6.8% to 84.6% and from 28% to 66% by bioclimatic zone and age group, respectively. Serological positivity was not statistically associated with gender. The presence of Leishmania DNA in blood, using PCR, revealed 21.2% (64/302) prevalence in dogs, which varied by bioclimatic zone (7.3% to 31%) and age group (7% to 25%). The entomological survey carried out in the studied localities showed 16 species of the two genera (Phlebotomus and Sergentomyia). P. perniciosus, P. papatasi, and P. perfiliewi were the most dominant species with relative abundances of 34.7%, 25% and 20.4%, respectively.Conclusions/SignificanceThe present report suggests a significant increase of CanL in all bioclimatic areas in Tunisia and confirms the ongoing spread of the infection of dogs to the country’s arid zone. Such an expansion of infection in dog population could be attributed to ecological, agronomic, social and climatic factors that affect the presence and density of the phlebotomine vectors.

Is it possible to consider the control of human Visceral Leishmaniasis from the perspective of Biomimetics?

Is it possible to consider the control of human Visceral Leishmaniasis from the perspective of Biomimetics?

Increased levels of cortisol are associated with the severity of experimental visceral leishmaniasis in a Leishmania (L.) infantum-hamster model.

BackgroundSeveral infectious diseases are associated with hypothalamic-pituitary-adrenal (HPA) axis disorders by elevating circulating glucocorticoids (GCs), which are known to have an immunosuppressive potential. We conducted this study in golden hamsters, a suitable model for human visceral leishmaniasis (VL), to investigate the relationship of Leishmania (L.) infantum infection on cortisol production and VL severity.MethodsL. infantum-infected (n = 42) and uninfected hamsters (n = 30) were followed-up at 30, 120, and 180 days post-infection (dpi). Plasma cortisol was analyzed by radioimmunoassay and cytokines, inducible nitric oxide synthase (iNOS), and arginase by RT-qPCR.ResultsAll hamsters showed splenomegaly at 180 dpi. Increased parasite burden was associated with higher arginase expression and lower iNOS induction. Cortisol levels were elevated in infected animals in all-time points evaluated. Except for monocytes, all other leucocytes showed a strong negative correlation with cortisol, while transaminases were positively correlated. Immunological markers as interleukin (IL)-6, IL-1β, IL-10, and transforming growth-factor-β (TGF-β) were positively correlated to cortisol production, while interferon-γ (IFN-γ) presented a negative correlation. A network analysis showed cortisol as an important knot linking clinical status and immunological parameters.ConclusionsThese results suggest that L. infantum increases the systemic levels of cortisol, which showed to be associated with hematological, biochemical, and immunological parameters associated to VL severity.

Splenic Transcriptional Responses in Severe Visceral Leishmaniasis: Impaired Leukocyte Chemotaxis and Cell Cycle Arrest.

Structural changes in the spleen have been reported in several infectious diseases. In visceral leishmaniasis (VL), a severe parasitic disease caused by Leishmania spp., the loss of white pulp accompanies a severe clinical presentation. Hamster model reproduces aspects of human VL progression. In the early stages, a transcriptomic signature of leukocyte recruitment was associated with white pulp hyperplasia. Subsequently, impaired leukocyte chemotaxis with loss of T lymphocytes in the periarteriolar lymphoid sheath occurred. This differential gene expression was subsequently corroborated by transcriptomic profiling of spleens in severe human VL. At the latest stage, spleen disorganization was associated with increasing clinical signs of VL. White pulp disruption was accompanied by decreased DLK1 expression. The expression of CXCL13, CCR5, CCL19, CCR6, CCR7 and LTA decreased, likely regulated by CDKN2A overexpression. Our findings enlighten a pathway implying cell cycle arrest and decreased gene expression involved in spleen organization.